Abstract

Acute stroke is a leading cause of disability and death, affecting over 1 million people per year in Europe alone. Physical activity is the most effective way to reduce the risk of all cardiovascular events, including stroke. However, after menopause, a transition where the risk of severe cardiovascular events increases drastically for women, the effects of physical activity on the vascular system are diminished. The molecular pathways underlying these important changes are not yet understood. To elucidate these pathways, we look specifically at noncoding RNAs. RNAs respond rapidly to stimuli such as physical activity and can travel and communicate between different tissues, such as between skeletal muscles and the brain. Furthermore, in response to such stimuli, RNAs undergo chemical modifications at individual nucleotide level, adapting the distinct biological functions of these RNAs. Similarly, factors like aging, changes in female hormone signaling and physical activity can modify our DNA, changing accessibility of certain regions of the genome and altering gene expression, including that of noncoding RNA genes. The ‘epitranscriptome’ and ‘epigenome’ together may hold the key to how physical activity can impact neurovascular health and modulate the risk of acute stroke after menopause. Combining existing studies on acute stroke in women with a new study that follows 200 women of varying physical activity/fitness levels going through menopausal transition, provides us with a unique set of human samples and data. In vitro studies on primary human brain endothelial cells are used to elucidate the underlying cellular RNA signaling pathways. With our findings we aim to empower women of all ages to minimize their risk of acute stroke via physical activity in an optimal manner and provide openings for research into other forms of cardiovascular diseases as well.

Abstract

My research career has taken me on a diverse journey, beginning with a solid foundation in basic hematology and expanding into a variety of interrelated disciplines. This early focus provided critical insights into cellular mechanisms and physiological responses, which led me to investigate environmental physiology and the adaptive strategies organisms employ in response to environmental stressors. The transition into eco-nanotoxicology marked a pivotal point in my career, where I explored the effects of nanomaterials on biological systems. This work illuminated the complexities of nanoparticle interactions within ecosystems and their implications for human health. Building on this foundation, I embraced computational biology and systems biology, utilizing these approaches to achieve a more holistic understanding of biological data. The integration of artificial intelligence (AI) tools brought innovative insights in understanding of biological responses to environmental challenges. My current projects focus on applying AI to interpret nanotoxicity data, bridging the gap between experimental findings and predictive modeling. Interdisciplinary integration in addressing the multifaceted challenges within biology and environmental science today is the future of biological research.

Abstract

Dr. Edi Prifti’s research lies at the intersection of bioinformatics and artificial intelligence, with a focus on predictive health applications. His work in microbiome metagenomics leverages the “Predomics” method, a novel approach aimed at predicting health outcomes by analyzing the complexities of microbial communities. By interpreting microbiome data with high precision, Predomics enables a deeper understanding of the microbiome’s role in human health, paving the way for targeted interventions. In parallel, Dr. Prifti is pioneering AI applications in cardiology through advanced analysis of electrocardiogram (ECG) data to predict sudden cardiac death risk. His work emphasizes developing robust classification models that balance accuracy with interpretability, ensuring that AI predictions are both reliable and transparent for clinical use. A central goal of his research is to enhance model robustness and interpretability, key factors for creating dependable AI solutions in healthcare. This dual focus on microbiome and ECG analysis highlights Dr. Prifti’s commitment to advancing AI-driven tools that inform preventive health strategies and support precision medicine, offering a blueprint for AI’s role in transforming healthcare.

Abstract

Although a small country, Albania shelters a wealth of aquatic and terrestrial ecosystems, in marine, coastal, hilly, and mountainous zones. Due to these circumstances, as other Mediterranean countries Albania is recognized as an important biodiversity hotspot in Europe, with rare and endangered habitats and species. Albanian nature has guaranteed survival and prosperity for humans over the centuries, in agriculture, animal husbandry, forestry, fishing, etc. Today, it is one of the strong points of tourism development, and it is often fascinating for many foreign visitors. A review of biodiversity values and their importance for man and nature will be emphasized here, and how to strengthen the friendly coexistence of man and nature. The friendly balance between development, conservation, and sustainable use of natural resources has not been easy in the new Albania's last 70-80 years. Despite the efforts towards the protection and related legal acts, nature and the natural resources are seriously impacted, especially in the last 30 years of the economic transition; the Protected Areas (PAs) are not saved either. Lastly, the Parliament, through Law 21/2024 amended Law 81/2017 on PAs; after that, all PA categories, even in their core areas can be developed for " infrastructure and hospitality activities, with the highest architectural and environmental standards for supporting tourism of excellence (with 5 stars) and the related infrastructure; the PA zoning was repealed too. All are not in the view of the nature protection, sanctioned in Albanian and international legal acts; but most of all, the approach is against the quality of life for the natural habitats, biota, and the human itself, and reducing the tourist values. Richness in habitats and plant and animal species is an important resource, from both natural and economic perspectives. However, it also implies the responsibility to preserve nature, habitats, and wildlife continuously. Careful use, preservation, and continuous restoration are the country's responsibility. It also means that ecosystem services must be always properly considered, as recognized by EU policy and international environmental conventions on the environment. How to strengthen the friendly coexistence of man and nature should be always a challenge, to Albania. Therefore, we must never stop efforts towards better understanding it, through education, awareness increase, through practical and concrete actions for an ecological society and ecological governance.

Keywords: Albanian natural values; PAs; Man and nature coexistence; Sustainable development; Ecological awareness.

About

This workshop will explore open science's principles, benefits, and practical implementation. Participants will learn how open access, open data, and open methodologies can enhance research transparency, reproducibility, and collaboration across the globe. The workshop will also provide an opportunity to learn the main concepts of research ethics and academic writing.  

About

Our genome is 3.055 billion-base pairs with millions of these base pairs differing between people, referred to as genetic variation. Understanding how this genetic variation relates to human disease and health requires statistic and computational methodologies. In this workshop we will cover some of the foundations of the field of statistical genetics including genotype imputation, genome wide association studies, burden tests, and polygenic risk scores.  

About

Effectively exploring and visualizing omics data is one crucial step for uncovering new biological insights. iSEE, a Bioconductor package/Shiny App, offers a wealth of powerful features to help you visualize your data at any stage of the data analysis pipeline. This workshop will cover some of the core functionalities of iSEE, as well as introducing iSEEfier, a Bioconductor package and iSEE extension, designed to simplify the setup of iSEE instances, making it easier to create customized visualization sessions. The participant will learn how to use iSEE and how to create tailored initial states with iSEEfier for diverse data exploration goals. 

About

 This workshop will introduce in one frame and practical terms the logic of sustainable development, its goals, and the role of education in sustainable development. The workshop will provide a practical approach to connect environmental themes and behaviors with sustainable development goals and increase understanding of how our institutions can be Eco-institutions and how our little daily actions can be translated into contributions to a sustainable society.   

About

We all know that doing a PhD is difficult at times but often we don’t feel comfortable to talk about the issues that we face. In this workshop, we would like to invite you to do just that. In addition to small-group and plenary discussions, participants will learn about potential ways to deal with difficulty in a functional way. We will focus on self-compassion as a tool that has been proven to be effective in boosting PhD students’ well-being.  

Abstract

Background and aim: Sexual reproduction of Toxoplasma gondii, which is restricted to the small intestine of felids, is sparsely documented, due to ethical concerns surrounding the use of cats as model organisms. Chromatin modifiers dictate the developmental fate of the parasite during its multistage life cycle, but their targeting to stage-specific cistromes is poorly described. 

Methodology: In this study, via inducible knock-downs (iKD), we found that transcription factors AP2XII-1 and AP2XI-2, expressed in tachyzoite stage that causes acute toxoplasmosis, can silence genes necessary for merozoites, a developmental stage critical for sexual commitment and transmission to the next host, including humans. Their conditional and simultaneous depletion leads to a drastic change in the transcriptional program, promoting a complete transition from tachyzoites to merozoites. 

Results: Pre-gametes produced in vitro under these conditions are characterized by specific protein markers and undergo typical asexual endopolygenic division cycles. In tachyzoites, AP2XII-1 and AP2XI-2 bind DNA as heterodimers at merozoite promoters and recruit the epigenitors MORC and HDAC31, which in turn restrict the accessibility of chromatin to the transcriptional machinery. Thus, the commitment to merogony stems from a profound epigenetic rewiring orchestrated by AP2XII-1 and AP2XI-2. 

Conclusion & future research: This effective in vitro culture of merozoites paves the way to explore Toxoplasma sexual reproduction without the need to infect kittens and has potential for the development of therapeutics to block parasite transmission.

Keywords: Toxoplasma gondii, sexual commitment, epigenetics, Apetala 2 transcription factors, merogony

Abstract

Background and aim: Anxiety disorders disproportionally affect females and are frequently comorbid with eating disorders. The emerging field of nutritional psychiatry has placed a particular focus on the impact of diet quality on anxiety pathophysiology, and the underlying mechanisms of gut microbiome function. A better understanding of how gut bacteria mediate and moderate such relationships could further contribute to developing personalized programs and inform probiotics and prebiotics manufacturing. To date, studies that look simultaneously at diet, the gut microbiome, and anxiety are missing as only pairwise relationships among them have been investigated. While the relationship between diet and anxiety is bidirectional, improving dietary habits could enhance the effects of pharmacological and psychological therapies.

Methodology: The present work used a deductive research methodology to summarize and integrate the  existing knowledge on the dietary effects on anxiety with focus on gut microbiome and with a sex-specific approach. Findings on the effects of diet on anxiety are critically summarized and reinterpreted in relation to findings on (i) the effects of diet on the gut microbiome composition, and (ii) the associations between the abundance of certain gut bacteria and anxiety.

Results:  Findings from this work suggest a theoretical model wherein the relationship between diet and anxiety is mediated and/or modulated by the gut microbiome through multiple mechanisms. In parallel, the present work further suggests that integrating multiple dietary assessment approaches and improving operationalization strategies could provide a more comprehensive understanding of how diet influences anxiety risk.

Conclusion & future research: The present research provides an update on the current state of nutritional research on anxiety while pinpointing methodological gaps that need further improvement. It also identifies gut microbiome genera with potential roles as mediators and proposes a new theoretical framework to guide future studies in this field.

Keywords: diet, diet quality, anxiety, gut microbiome, females

Abstract

Background and Aim: Although the importance of a healthy microbiome and the abundance of microorganisms in the gut is well recognized, precise quantitative data on the microbial biomass in the human large intestine is lacking. Current information is limited to estimates not based on experimental measurements of native biomass (Sender et al., 2016). Additionally, the extent of microbial contribution to stool composition and the effect of health conditions on total microbial biomass remains unclear. In my master’s thesis, we aim to establish a protocol to preserve both microbial cell mass and cell count over time and address key questions: How do fixatives impact cell weight and cell count over time? Is there intra-sample and inter-sample variability in cell masses and counts over 1- to 2-week intervals. Further, we want to see if vegetarian and omnivorous diets yield different microbial biomass. 

Methodology: We sampled 10 healthy volunteers (5 male, 5 female) at three different time points within a 1- to 8-week period. Three volunteers were vegetarians. Undigested organic material was removed by slow-speed centrifugation, followed by fixation in PFA or DSP. We then measured cell count by flow cytometer and buoyant single-cell mass by Life Scale (suspended micro-channel resonator combined with picoliter-scale microfluidic control) and used these values to estimate dry microbial mass. 

Results: We determined that cell mass remains stable over one month when using DSP or PFA fixation; however, cell count decreases significantly within 24 hours. Our findings suggest that intra-sample variability sometimes exceeds inter-sample variability, indicating that stool consistency, as categorized by the Bristol Stool Chart, may play a significant role. Additionally, we observed higher inter-sample variability in vegetarians compared to omnivores. 

Conclusion & Future Research: Our preliminary data indicate that microbial dry mass in human stool ranges from 0.006 g to 0.03 g per gram of stool, establishing a baseline for microbial biomass in a healthy population. The stability of cell mass with DSP and PFA fixation over one month suggests that these protocols are effective for longitudinal studies, although cell count stability remains a limitation. The observed variability based on stool consistency highlights the importance of considering stool type when evaluating microbial biomass. Our next steps will involve analyzing existing data on participant gender, age, body weight, and dietary background to determine their influence on microbial biomass. Future research could focus on including individuals with diverse health conditions, such as gastrointestinal disorders, to investigate how microbial biomass may vary across health statuses.  

Keywords:  Gut microbiome, Microbial biomass, Dietary impact, suspended micro-channel resonator, Flow cytometry 

Abstract

Background and aim:  Chronic inflammatory diseases (CID), such as rheumatoid arthritis, are not only characterized by direct inflammatory responses but also by alterations in oxidative status and lipid metabolism, increasing the risk of atherosclerosis and cardiovascular mortality. This study aimed to evaluate the potential of astaxanthin (ASX), a natural compound, in normalizing inflammation-induced changes in the expression of inflammatory, antioxidant, and lipid-associated genes in the liver. 

Methodology: In vitro experiments were conducted using a model of HepG2 hepatocytes stimulated with THP-1 monocyte-conditioned medium (CM). Cells were pre-treated with varying concentrations of ASX for different durations, followed by gene expression analysis via qRT-PCR. In vivo studies involved the induction of adjuvant arthritis (AA) in rats through intradermal injection of Mycobacterium butyricum at the base of the tail. ASX was administered daily at doses of 1 mg/kg and 5 mg/kg. Clinical parameters in rats, specifically body weight and hind paw volume, were measured on a weekly basis. After 28 days, the rats were sacrificed, and liver samples were collected for qRT-PCR analysis. Antioxidant status was assessed in erythrocytes by measuring superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities, and in plasma by determining Trolox equivalent antioxidant capacity (TEAC) and lipoperoxide (LPx) levels. 

Results: In vitro, ASX had no significant effect on CM-induced changes in mRNA expression in HepG2 cells. The lack of ASX’s in vitro activity may be due to its poor water solubility, which likely limited its cellular uptake in the aqueous culture medium. Subsequent attempts to enhance ASX uptake through complexing with bovine serum albumin were not significantly successful. In vivo, no significant effect of ASX on clinical parameters nor hepatic expression of inflammatory or lipid-associated genes expression was observed, however, ASX modulated the hepatic expression of antioxidant genes, including heme oxygenase 1 (HO-1), CAT, and lipoprotein-phospholipase A2 (Lp-PLA2) and improved antioxidant activity in erythrocytes and plasma. 

Conclusion & future research: ASX demonstrated efficacy in modulating liver antioxidant gene expression and plasma and erythrocyte antioxidant activity in vivo, but had no effect on inflammatory or lipid-associated genes and other clinical parameters, either in vivo or in vitro. Future in vitro studies should focus on enhancing ASX bioavailability, possibly through the development of nanoparticle-based drug delivery systems.

Keywords: inflammation, lipid metabolism, antioxidants, liver

Abstract

Background and aim: Unsupervised learning techniques can detect clinically relevant structure in a population cohort data of human gut microbiota. While the gut microbiota composition is influenced by individual factors such as diet, medication, and development of the immune system during early childhood, it is proposed that individuals maintain a relatively stable microbiota ecosystem throughout adulthood. This stability allows to distinguish individuals into subgroups based on their gut microbiota characteristics, which leads us to the objective of this work: to comprehensively study techniques for low-resolution microbiota community typing enabling the definition of the key characteristics of common microbiota assemblages in the Finnish population.

Methodology: We compared three probabilistic unsupervised learning techniques, non-negative matrix factorization, Dirichlet Multinomial Mixtures and Latent Dirichlet Allocation, with a naive benchmark, using an all-cause mortality association strength as a quantitative metrics, to distinguish biologically relevant structure in a large Finnish population cohort, FINRISK, with almost 18 years follow-up. The techniques identify microbiota assemblages as either discrete enterotypes, which assign each sample into a single community type, or continuous enterosignatures, which identify patterns of co-occurrence of microbiota community types within each sample. Tools for microbiome analysis were provided by the mia package in Bioconductor, utilizing the TreeSummarizedExperiment infrastructure.

Results: We found five rather robust community types, characterized by Bacteroides, Alistipes, Agathobacter, Escherichia, and Prevotella bacterial genera. All techniques uncovered clinically relevant structure in the data, yet Latent Dirichlet Allocation detected the strongest early mortality signal using Cox regression, outperforming all other techniques. The replicability of Latent Dirichlet Allocation was assessed using cross-validation. The predicted community types uncovered similar ecological landscape on the data with the community types obtained using the entire data, confirming the clinical relevance, robustness, and scalability of the technique.

Conclusion & future research: Such findings emphasize the relevance of this work in advancing both our understanding microbiota communities in human population and the methods suitable to analyse these communities. Comparison of the unsupervised microbiota community typing techniques builds ground for further studies of such ecosystems in more detail, as the choice of the unsupervised model characterizes the learning process. From these findings, we can conclude that Latent Dirichlet Allocation is an effective technique for the large-scale microbiota community typing. Further exploration of this result has the potential to offer valuable insights into the ecological interactions within the microbiota communities and their impact on disease development. 

Keywords: microbiota, unsupervised, latent feature, probabilistic, Bayesian, survival 

Abstract

Background & Aim: Gliomas are aggressive brain tumors difficult to treat mostly due to their large molecular heterogeneity. This requires continuous improvement in the molecular characterization of the glioma types to identify potential therapeutic targets. Advances in glioma research are rapidly evolving, contributing to the updates of the WHO classification of tumors. Data analysis of multiple omics layers through classification and feature selection methods holds promise in identifying crucial molecular features for distinguishing between glioma types. 

Methodology: We developed a robust and sparse classification workflow based on multinomial logistic regression to investigate the molecular landscape of gliomas. We considered transcriptomics and methylomics glioma profiles of patients labeled following the latest WHO glioma classification updates (2016 and 2021).

Results: Overall, our results show a notable improvement in glioma types separability for the 2021 WHO updated patient labels at both omics levels. Patients flagged as outliers for the 2016 WHO classification exhibited a molecular profile deviating from the one of the respective classes, which was more aligned with the current associated glioma type according to the 2021 WHO update.

Conclusion & future work: The methylomics profiles were particularly promising in the identification of outliers. These contributions will support further revisions of glioma molecular characterization and the development of novel targeted therapies.

Keywords: Glioma, Omics, Feature selection, Multinomial classification, Outlier detection.

Abstract

Background and aim: Deciphering the underlying mechanisms driving cancer progression and therapeutic resistance is important for improving patient outcomes and developing effective treatment strategies. Transcriptomics uncovers key regulatory pathways and molecular signatures associated with disease progression. Meanwhile, metabolomics provides a snapshot of cancer metabolic phenotype, highlighting metabolic reprogramming and dysregulated pathways driving cancer growth and survival. However, the direct linkage of transcriptomic and metabolomic levels of regulation is still challenging. In this study, we present an integrative analysis of transcriptomics and metabolomics data modalities based on matrix factorization. 

Methodology: Our investigation utilizes scRNA-seq data from NRAS-mutated melanoma cell lines treated with MEK1/2 plus CDK4/6 inhibitors, bulk RNA-seq data from open access GBM dataset, and metabolomics data for both cancer types. Leveraging advanced computational methods, including consensus Independent Component Analysis (consICA) and machine learning approaches, we integrate these omics datasets to extract molecular signatures associated with cancer progression.

Results: We investigated TCA cycle activation and linked enriched pathways with metabolite level changes. Predicting models for metabolites were built on transcriptomics data. Facing the limitations of the predictive approach, we are currently working on validation using metabolomics experiments. Additionally, we observed the limitations in linking transcriptomics to metabolomics using flux analysis and characterized the robustness of published models such as scFEA. Finally, for melanoma single-cells, we established cell cycle stages and observed a strong linkage between the proportion of proliferating cells and adaptation to the treatment, which occurred at individual speed for each cell line. 

Conclusion & future research: The integrative analysis reveals that combining transcriptomics and metabolomics using matrix factorization approaches, such as consICA, provides valuable insights into cancer progression and treatment adaptation. The observed associations between transcriptomic changes and metabolite variations underscore the complex interplay between gene expression and metabolic reprogramming in cancer. Future work will involve experimental validation of transcriptome-based predictions for metabolite levels Additionally, a deeper investigation into cell line-specific adaptive responses at the single-cell level will aim to advance personalized therapeutic strategies.

Keywords: cancer biology, transcriptomics, metabolomics, integrative analysis, ICA.

Abstract

Background and aim: With the exponential growth in the size and complexity of single-cell RNA-seq data, traditional workflows are becoming increasingly computationally intensive. Existing tools often need help to efficiently handle such large datasets, frequently running out of memory. To address this challenge, the adoption of more efficient algorithms and out-of-memory data representations is crucial for effective analysis.

Methodology: This study presents a comprehensive comparison of various workflows for single-cell data analysis, evaluating their performance and efficacy within both R and Python programming environments. Specifically, we assess the capabilities of Seurat and Bioconductor in R, and Scanpy and rapids_singlecell in Python, with a focus on their utilization of both Central Processing Units (CPUs) and Graphics Processing Units (GPUs) for optimising computational efficiency. Utilising a diverse set of real single-cell RNA-seq datasets, including approximately 1.3 million cells from the mouse brain, BE1, sc_mixology, and cord blood datasets, we expanded the benchmarking to encompass the entire workflow of single-cell analysis. This comprehensive evaluation enabled us to assess the scalability of the workflows across different datasets. Furthermore, recognizing the need for optimized workflows within the Bioconductor framework to handle large single-cell RNA-seq data, we developed a vignette featuring the most efficient methods in terms of scalability, computational time, and memory usage. This vignette serves as a valuable resource for researchers working with large single-cell RNA-seq datasets within the Bioconductor ecosystem. In addition to evaluating computational efficiency, we assessed the concordance between workflows by comparing the Rand index using cell annotations and cluster analysis. This comparative analysis provided insights into the consistency and accuracy of the workflows in identifying cellular groups and their annotations.

Results: The results revealed significant disparities in computational times across workflows and datasets, with GPU-accelerated approaches consistently outperforming CPU-centric methods. Moreover, the comparison of Rand indices demonstrated high concordance between workflows, affirming their reliability and consistency in identifying cellular groups across diverse datasets.

Conclusion & future research: Overall, this research underscores the importance of efficient computational frameworks in handling the escalating demands of single-cell RNA-seq data analysis. The findings not only offer valuable insights into selecting the optimal workflow based on trade-offs between time and memory consumption but also emphasize the transformative impact of GPU acceleration in expediting large-scale single-cell data analysis while ensuring robust and accurate results.

Keywords: benchmark, single-cell RNA-seq, GPU, CPU, Python, R. 

Abstract

Background & Aim: Oligodendrocytes are cells of the central nervous system that form myelin around neuronal axons and modulate how quickly electrical signals travel throughout the brain. These cells have a readily available pool of resident progenitors called oligodendrocyte precursor cells (OPCs). Oligodendrocyte generation from OPCs persists throughout life and allows for oligodendrocyte and myelin replacement in neurodegeneration and aging. The cellular and molecular mechanisms that guide certain subpopulations of OPCs to generate oligodendrocytes are not fully understood. Intracellular metabolism and organelle dynamics are likely to play key roles in regulating this decision. 

Methodology: Using high-resolution intravital optical imaging techniques to trace fluorophore labeled-oligodendrocyte lineage cells and their mitochondria, we determined how mitochondrial distribution, shape, and dynamic movement throughout the cell change as single OPCs transform into myelinating oligodendrocytes. 

Results: We showed that during oligodendrocyte generation, mitochondrial content expands concurrently with a change in subcellular partitioning towards the distal processes. These changes are followed by an abrupt loss of mitochondria in the oligodendrocyte processes and myelin, coinciding with sheath compaction. This reorganization and extensive expansion and depletion take 3 days. In addition, oligodendrocyte mitochondria are stationary over days while OPC mitochondrial motility is modulated by animal arousal state within minutes. Aged OPCs also display decreased mitochondrial size, volume fraction, and motility. 

Conclusion: Mitochondrial dynamics are linked to oligodendrocyte generation, dynamically modified by their local microenvironment, and altered in the aging brain.

Keywords: oligodendrocyte, myelin, mitochondria, differentiation

Abstract

Background and aim: Molecular therapies targeting the EGFR/MAPK pathway have significantly progressed in colorectal cancer (CRC) treatment. However, primary resistance and the emergence of secondary resistance stemming from either pre-existing or adaptive mechanisms continue to pose major challenges. In cancer cells, oncogenic signaling often activates stress response pathways, enabling these cells to manage the stress induced by oncogenes. The hyperactivation of these pathways may expose inherent vulnerabilities that could be targeted for therapeutic purposes. We aim to overcome primary and secondary resistance to targeted therapies in CRC by harnessing the potential of DNA damage response inhibitors (DDRi-s) as a novel therapeutic strategy.

Methodology: To address primary resistance we leveraged our unique biobank of 226 CRC cell lines, selecting 112 models representing the genomic landscape of metastatic CRC. These models were challenged with various DDRi-s targeting replication stress (RS), homologous recombination (HR) repair, and non-homologous end-joining (NHEJ) repair pathway. We mainly focused our attention on inhibitors that target the replication stress pathway, specifically ATR and WEE1 inhibitors, which showed the greatest efficacy. To identify potential biomarkers of response and resistance, we analyzed CRC models displaying high sensitivity or resistance to DDR inhibitors across multiple parameters. To address secondary resistance, we generated several unique CRC models with acquired resistance to different drugs targeting the EGFR/MAPK pathway (ARes platform) and evaluated their sensitivity to DDR inhibition following resistance acquisition.

Results: To address primary resistance, we observed that approximately 30% of colorectal cancers, including those with KRAS and BRAF mutations that are unresponsive to targeted agents, showed sensitivity to at least one DDR inhibitor within to the replication stress pathway. In our biomarker investigation for ATR inhibitor (ATRi) sensitivity, we observed that ATRi-sensitive cells had fewer phospho-RPA32 foci at baseline, while ATRi-resistant cells exhibited increased RAD51 foci formation in response to replication stress. Furthermore,  lack of ATM and RAD51C expression correlated with ATRi sensitivity. Additionally, our analysis of mutational signatures and the HRDetect score revealed a subgroup of models sensitive to ATRi. Notably, organoids derived from patients with metastatic colorectal cancer mirrored the findings obtained from cell lines. In the context of secondary resistance, replication stress and in particular WEE1 resulted as the most compelling targets both in vitro and in vivo.

Conclusion & future research: In conclusion, Our findings suggest that a subset of colorectal cancers resistant to current therapies may benefit from DDR pathway and replication stress inhibitors. A composite biomarker involving phospho-RPA32 and RAD51 foci, lack of ATM and RAD51C expression, and analysis of mutational signatures could aid in identifying colorectal cancers likely to respond to inhibitors of the replication stress pathway. Replication stress and WEE1, particularly, represent a valuable therapeutic targets in overcoming secondary resistance to EGFR-targeted therapies in CRC. In future studies, we plan to identify  alterations in the acquired resistant models that enhance susceptibility to WEE1 inhibition, establishing predictive biomarkers of therapeutic response.

Keywords: Colorectal Cancer, Resistance, DDR, Replication Stress, ATR, WEE1. 

Abstract

Background and aim: Calcium (Ca2+) act as essential messengers in inducing apoptosis, influencing various cellular events and leading to mitochondrial outer membrane permeabilization (MOMP). The Bcl-2 family proteins are key regulators of mitochondrial apoptosis. Anti-apoptotic Bcl-2 protein is well known for its non-canonical role in regulating the pro-apoptotic Ca2+ signals mediated by inositol 1,4,5-trisphosphate receptors (IP3Rs). Upon apoptotic stimuli, Bcl-2 suppresses the excessive efflux of Ca2+ from endoplasmic reticulum (ER) to mitochondria in mitochondria-associated membranes (MAMs). This is achieved by inhibition of IP3R via binding of BH4 domain of Bcl-2 to specific regions on the IP3R. The aims of this study are to (i) investigate the impact of Bcl-2 on ER-mitochondria Ca2+ dynamics, (ii) establish a novel system using mitochondrial targeted Yellow Cameleon (YC) sensors for direct measurements of mitochondrial Ca2+ upon overexpression of Bcl-2 protein and (iii) Optimizing a system using YC biosensore to measure interplay between Bcl-2 and cytosolic Ca2+. 

Methodology: To study the role of anti-apoptotic Bcl-2 protein in suppression of Ca2+ response at cytosolic and mitochondrial level, Bcl-2 protein was overexpressed in HeLa cells. Ca2+ levels in living cells overexpressing Bcl-2 protein were visualized using YC biosensor. Subsequently, ratiometric measurements of Ca2+ responses were obtained through confocal microscope. The data were statistically analyzed to conclude in the impact of Bcl-2 in Ca2+ response in HeLa cells.

Results: Bcl-2 reduces both the cytosolic and mitochondrial Ca2+ response.

Conclusion & future research: FRET- based Ca2+ sensors with distinct affinities are compatible for studying non-canonical Bcl-2 function. Bcl-2 suppresses agonist induced Ca2+ signaling in cytosol and mitochondria irrespective of the agonist concentration. The next step on the project would be to study the impact of Bcl-2 on the cytosolic and mitochondrial Ca2+ exchange upon directly inducing apoptosis on the cells. This can be investigated by using a different stimulator, such as staurosporine. Moreover, to further investigate the involvement of each three IP3R isoform, IP3R knockout and rescue can be performed in the future. 

Keywords: Ca2+, Yellow Cameleon biosensore, Bcl-2, mitochondria. 

Abstract

Background and aim: Variants in membrane trafficking proteins are known to cause rare disorders with severe symptoms. The highly conserved transport protein particle (TRAPP) complexes are key membrane trafficking regulators that are also involved in autophagy. Pathogenic genetic variants in specific TRAPP subunits are linked to neurological disorders, muscular dystrophies, and skeletal dysplasias. Characterizing these variants and their phenotypes is important for understanding the general and specialized roles of TRAPP subunits as well as for patient diagnosis. Patient-derived cells are not always available, which poses a limitation for the study of these diseases. Therefore, other systems, like the yeast Saccharomyces cerevisiae, can be used to dissect the mechanisms at the intracellular level underlying these disorders. 

Methodology: The development of CRISPR/Cas9 technology in yeast has enabled a scar-less editing method that creates an efficient humanized yeast model. In this study, core yeast subunits were humanized by replacing them with their human orthologs, and TRAPPC1, TRAPPC2, TRAPPC2L, TRAPPC6A, and TRAPPC6B were found to successfully replace their yeast counterparts. This system was used for studying the first reported individual with an autosomal recessive disorder caused by biallelic TRAPPC1 variants, a girl with a severe neurodevelopmental disorder and myopathy. 

Results: We show that the maternal variant (TRAPPC1 p.(Val121Alafs*3)) is non-functional while the paternal variant (TRAPPC1 p.(His22_Lys24del)) is conditional-lethal and affects secretion and non-selective autophagy in yeast. This parallels defects seen in fibroblasts derived from this individual which also showed membrane trafficking defects and altered Golgi morphology, all of which were rescued in the human system by wild-type TRAPPC1. 

Conclusion & future research: This study suggests that humanized yeast can be an efficient means to study TRAPP subunit variants in the absence of human cells and can assign significance to variants of unknown significance (VUS). This study lays the foundation for characterizing further TRAPP variants through this system, rapidly contributing to disease diagnosis. 

Keywords: yeast; TRAPP; TRAPPC1; mutation; humanization; Golgi; autophagy

Abstract

Background & Aim: Proteins are essential tools in biotechnology, enabling the production of therapeutics, vaccines, and diagnostic reagents. However, their utility is constrained by stability and solubility issues, potential immunogenicity, and altered biological activity. Over the years, various protein engineering methods have emerged to address these limitations, and one particularly promising approach is glycoengineering. Glycoengineering involves the manipulation and modification of the glycans attached to proteins or other molecules. Glycans play crucial roles in various biological processes, including cell signalling, immune response, and protein folding. By modifying these glycans, desired properties or functions can be achieved.  Since glycosylation is not template-driven, presence of specific glycotransferases and glycosidases is needed.  On the other hand, Mollusca is one of the most diverse groups of animal kingdom and they display significant functions in many ecosystems as they are important filter feeders and decomposers, but they are also pests in agriculture and intermediate hosts of many human and cattle parasites. Their ability to produce complex glycans is a fundamental aspect of their biology. In many cases molluscs display similar glycan structures with the human parasites they host. This makes them an interesting system to study glycosylation.   In this case, the identification of three novel core 1 β-1,3-galactosyltransferses (T-synthase) from Biomphalaria glabrata, Pomacea canaliculata and Crassostrea gigas presents exciting opportunities for biotechnological applications but also gives a better understanding of the glycosylation processes in molluscs and sheds light on their adaptation and survival abilities. 

Methodology: The coding sequences were identified by homology search synthesised and were expressed in Sf9 cells. The expression product of the putative T-synthases displayed core 1 β1,3-galactosyltransferase activity using pNP-α- GalNAc as the substrate. Biochemical characteristics and structural features were analysed and compared with previously characterised enzymes from other species. 

Results & Conclusion: In this study, we present the identification and characterization of novel T-synthases, providing insights into molluscan glycosylation abilities, expanding our knowledge of this enzyme across species, and offering new potential tools for biotechnological applications.  

Keywords: O-glycosylation, Mollusca, T-synthase, enzyme.

Abstract

The Vjosa River and its tributaries represent a large and dynamic fluvial network characterized by a longitudinal continuity in water flow which remained undisturbed throughout its length. Not surprising, the Vjosa River system is a hotspot of biodiversity. It is assumed that the river network provides essential ecosystem services, with a high self-purification capacity of exceptional importance for biodiversity and human needs. Due to its intact hydromorphology, the near natural flow and sediment regime, as well as the exceptional biodiversity, the Vjosa River is a role model of an intact natural river network. 

In rivers, contaminants are attenuated and degraded only to a minor extend in the visible river channels. Natural attenuation mainly takes place in the hyporheic zone (HZ) which is the interface between surface water and groundwater. The HZ is kind of a natural bioreactor, densely colonized by microorganisms and diverse invertebrates. 

This study focuses on key biogeochemical processes involved in the cycling of carbon and nutrients in a qualitative and quantitative manner. As one aspect, we investigate self-purification activities in the sediments of river gravel bars, which are considered hotspots for biogeochemical activity.  First, numerous piezometers were pushed into gravel bar sediments. Subsequently, the subsurface hydrology was assessed by means of conservative tracer (uranine, deuterated water) injection and monitoring of breakthrough curves. In the following, a natural DOM source (from leave leachate) was constantly injected accompanied by daily sampling of all piezometers for several days to weeks. Water samples collected were analyzed for basic physico-chemical parameters (temperature, pH, EC, DO), major ions and nutrients (NO3-, NO2-, NH4+, PO43-), concentration of DOC, DIC, and CO2, as well as the composition and quality of dissolved organic matter (DOM). Moreover, microbial biomass was collected for a later analysis of microbial community composition. 

Our findings, will contribute to a better understanding of the natural attenuation potential in an ecologically intact river ecosystems and will highlight the importance of the Vjosa River as a unique natural laboratory and reference system for other rivers in Europe. 

Keywords: natural attenuation, hyporheic zone, gravel bar, microbial community

Abstract

Background and aim: Albania is considered one of the countries with the highest percentage of untreated plastic waste. Microplastic studies represent a significant gap in environmental research in Albania, particularly regarding marine ecosystems. This study aims to provide an overview of microplastic pollution in Vlora Bay, focusing on their distribution in sediments, water columns, and fish intestines, while proposing strategies for mitigation. 

Methodology: In a collaboration between the Agricultural University of Tirana and the University of Camerino, four sites in Vlora Bay, including the marine protected area of Karaburun-Sazani, are being analyzed. Methodologies include Manta net use for water columns sampling, direct sampling of coastal sediments, direct collection of plastics scattered in the coast and studying of the microorganisms associated with the plastics. Finally, we established a collaboration with small-scale fishers for collecting specimens of the common sole (Solea solea). The samples for the microplastic – associated microorganisms are collected in three replicas per site and in a random collection of plastics along the coast, while microplastics characterization also includes the sampling with the Manta net. The methodology of DNA extraction with quantification protocols, has been established to assess microplastic-associated microorganisms via metagenomics, while the characterization of the microplastics will be conducted with FTIR&RAMAN Spectroscopy in collaboration with the Chemistry Department of University of Camerino. 

Results: Preliminary assessments conducted in October 2023 and February 2024 identified microplastics in seabed  sediments, water samples, and fish intestines. The sampling season in July and early September 2024 produced results regarding the microbial colonization and the presence of microplastics in the site. The results are still to be elaborated regarding the characterization of the microplastics.

Conclusion & future research: These preliminary findings underline the urgency of addressing plastic waste in Albania. Planned samplings for June and August 2025 and 2026 aim to refine the understanding of microplastic presence and their ecological impact, in order to propose mitigation strategies.

Keywords: microplastics, marine ecosystems, Vlora Bay, Karaburun-Sazani, plastic pollution mitigation, metagenomics

Abstract

Background & Aim: Tirana, the capital of Albania, is the most urbanized area in the country, boasting a population exceeding 1 million inhabitants. Research on the diversity and other aspects of urban flora are limited in Albania, despite the significance of such ecosystems. This study investigates the diversity and characteristics of the urban flora in Tirana. 

Methodology: The field trips for plant collections were conducted in 20 randomly selected sampling areas. The statue of Scanderbeg was taken as the central point. A circular area with a radius of 2 km had been selected from this statue. Grids of 250 m x 250 m squares were drawn on this layer and within these grids, 20 were selected and sampled using the random selection function in the QGIS 3.4 Madeira program. 

Results: Our research into the urban areas selected for this study reveals the presence of 317 native taxa. This diverse group of plants is primarily characterized by its spontaneous species, which account for approximately 275 taxa. These spontaneous natives have adapted to their local environments, thriving in various urban habitats and contributing to the overall biodiversity of the area. In contrast, only 42 of the native taxa are cultivated as ornamental plants, such as Prunus cerasifera Ehrh., Aesculus hippocastanum L., Tilia tomentosa Moench, etc. In the urban ecosystems of this study, alien flora plays a significant role, comprising a total of 294 distinct taxa. This diverse array of plant species can be categorized into two main groups: cultivated alien flora and spontaneous alien flora. The majority of alien plants within urban settings are cultivated species, totaling 253 taxa. These plants are primarily introduced for ornamental purposes, enhancing the aesthetic appeal of gardens, parks, and public spaces. In contrast, spontaneous alien flora consists of 41 taxa that have established themselves naturally in urban environments, such as Alopecurus utriculatus Sol., Dichondra micrantha Urb., Commelina communis L., etc.  These species often thrive in disturbed soils and can adapt to the unique conditions found in cities. While they may not be intentionally planted, they contribute to the biodiversity of urban landscapes and can play a role in ecological processes. 

Conclusion: Studying the urban flora of a city is vital for conserving native plant species, enhancing ecological health, and promoting sustainable practices that improve the overall quality of urban life.

Keywords: Urban flora, Tirana, native, alien, spontaneous, cultivated.

Abstract

Vitamin D3 (VD3) is a fat-soluble prohormone synthesized primarily in the skin through exposure to ultraviolet B (UVB) rays from sunlight, as well as absorbed from dietary sources. Traditionally, VD3 has been recognized for its crucial role in calcium homeostasis and bone metabolism. However, recent research has uncovered its broader physiological effects, particularly within the central nervous system (CNS), where VD3 exerts neuroprotective functions. The hippocampus, a critical brain structure involved in learning, memory, and cognitive function, is notably responsive to VD3 due to the presence of Vitamin D receptors (VDRs) in hippocampal neurons. This study aimed to explore the effects of VD3 on the growth and differentiation of hippocampal embryonic cells from the HN9.10E cell line, focusing on the morphological changes in neuronal structures.

The research was conducted as part of the Erasmus+ program in collaboration with the University of Tirana and the University of Perugia. The experimental design involved culturing HN9.10E embryonic hippocampal cells, which were divided into two groups: a control group without VD3 treatment and a treatment group exposed to VD3. VD3 was administered at various concentrations to observe potential dose-dependent effects. Cells were monitored over 24, 48, and 72 hours, and the morphological parameters such as soma size, dendritic length, and general cellular morphology were assessed using microscopic imaging and morphometric analysis. Quantitative measurements were then compared between the two groups to determine the impact of VD3 on neuronal development. Statistical analysis was applied to assess the significance of the findings.

The study demonstrated that VD3 significantly enhanced the growth and differentiation of hippocampal embryonic cells compared to the control group. Specifically, VD3-treated cells exhibited larger soma sizes and longer dendritic extensions, indicating increased neuronal differentiation and maturation. These morphological changes were particularly evident after 48 hours of treatment and were more pronounced at higher VD3 concentrations, suggesting a dose-dependent relationship. Moreover, the findings suggest that VD3 may promote neurogenesis through interactions with neurotrophic factors such as Nerve Growth Factor (NGF) and Brain-Derived Neurotrophic Factor (BDNF), both crucial for neuronal survival, growth, and synaptic plasticity. The ability of VD3 to influence these key cellular processes highlights its importance in maintaining hippocampal health and function.

The results of this study support the hypothesis that VD3 plays a neuroprotective and neurogenic role in hippocampal cells. By promoting neuronal growth and differentiation, VD3 may enhance hippocampal function, which is vital for memory formation and cognitive health. This has significant implications for understanding how VD3 deficiency may contribute to cognitive decline and neurodegenerative diseases such as Alzheimer’s disease, where hippocampal dysfunction is a hallmark. Given the growing evidence of VD3’s involvement in brain health, further research should investigate the molecular mechanisms underlying VD3’s effects on hippocampal neurons. Future studies should also explore the potential therapeutic applications of VD3 supplementation in clinical settings, particularly in the prevention and treatment of neurodegenerative conditions. Additionally, longitudinal studies are needed to assess the long-term impact of VD3 on hippocampal function and cognitive outcomes, and to explore its interactions with other neurotrophic factors.

Keywords: Vitamin D3, hippocampal neurons, neurogenesis, neuronal differentiation, neurotrophic factors, in vitro study.

Abstract

Background and aim: Tobacco consumption has a significant impact on human health. Recent studies have also demonstrated that smoking induces changes in the gut microbiome. This study aims to investigate the effects of smoking on the human gut microbiome and explore the link between smoking-induced microbiome alterations and differential immune responses upon stimulation with various antigens. 

Methodology: Whole genome shotgun sequencing (WGS) data was obtained from the Human Functional Genomics Project1. The dataset includes fecal samples from 60 current smokers and 339 healthy individuals who had never smoked, all without diseases affecting gut flora. Alpha and Beta diversity analyses were performed using the vegan package2. Differential abundances between smokers and non-smokers were analyzed using SIAMCAT3, which was also used to train and assess random forest models. Additionally, metabolomics and immunological datasets from blood serum samples of the same cohort were integrated using canonical correlation analysis (CCA) with MintTea4.

Results: Six microbial species exhibited differential abundance between smokers and non-smokers after filtering for taxa present in at least 10% of all samples. These findings were consistent with prior literature. However, a random forest model showed only limited ability to classify smoking habits based on metagenomic inputs (average accuracy = 0.55).. MintTea identified three biological modules associated with smoking among all input features. One module includes Ruminococcus gnavus, which was also the most significantly altered species in this study between smoking subgroups. Furthermore, cholesterol levels from serum metabolomics and TNFɑ levels from immunological measurements are part of the same module, suggesting a potential connection between these factors.

Conclusion & future research: Emerging research highlights the connection between the gut microbiome, the brain, emotional well-being, and the immune system. This study shows that Smoking habits do alter the human gut microbiome, though the changes appear to be subtle. The findings here suggest that smoking may influence the immune system and human well being via microbiome alterations, though further research is necessary to elucidate the role of e.g. Ruminococcus gnavus or other species in this process.

Keywords: smoking, gut microbiome, metagenomics, canonical correlation analysis, data integration

1. Human Functional Genomics Project. http://www.humanfunctionalgenomics.org/site/ 

2. Oksanen, Jari et al. (2024). vegan: Community Ecology Package. URL : https://github.com/vegandevs/vegan

3. Wirbel J, et al., Genome Biol 22, 93 (2021). https://doi.org/10.1186/s13059-021-02306-1

4. Muller, E, et al.,. Nature Communications, 15,1 (2024). https://doi.org/10.1038/s41467-024-46888-3 

Abstract

Background and aim: Nf2/Merlin, a member of the ERM protein family, plays a pivotal role in cell cortex integration, plasma membrane function, and cytoskeletal stability. Osteoarthritis (OA) is a degenerative joint disease characterized by the breakdown of articular cartilage and associated with global disability. Although Nf2/Merlin is present in cartilage, its role in OA progression is underexplored. This study investigates the involvement of Nf2/Merlin in regulating cartilage homeostasis and its potential as a therapeutic target for OA treatment.

Methodology: The study used a mouse model of OA induced by surgical destabilization of the medial meniscus (DMM). Both control (Nf2-NC) and knockout (Nf2-cKO) chondrocytes were cultured from articular cartilage and meniscus tissues. Transcriptome sequencing was performed to analyze gene expression changes, focusing on signaling pathways related to cartilage development. Micro-CT was used to assess structural changes in subchondral bone, while IHC and qPCR were conducted to validate the expression of inflammatory markers such as MMP13, IL-1β, and IL-6 in Nf2-deficient cartilage.

Results: Nf2-cKO samples showed significant alterations in genes regulating cell adhesion, signaling, and extracellular matrix composition. Micro-CT analysis revealed decreased sclerosis, BV/TV, and increased BMD in knockout mice. IHC and qPCR analyses confirmed the upregulation of inflammatory mediators, suggesting that Nf2/Merlin deficiency modulates key processes involved in OA progression.

Conclusion & future research: Nf2/Merlin plays a critical role in maintaining cartilage integrity by modulating gene expression and inflammatory responses. Its absence leads to structural changes and heightened inflammation in OA, making it a promising target for future therapeutic interventions aimed at managing OA progression. Further studies should explore the potential of Nf2/Merlin as a biomarker and therapeutic target in clinical settings.

Keywords: Osteoarthritis, Nf2/Merlin, cartilage homeostasis, gene knockout, inflammation, signaling pathways. 

Abstract

Background and aim: Cells organize their intricate biochemical processes into specialized compartments responsible for various molecular functions. Recent research suggests that intrinsically disordered proteins (IDPs) are key orchestrators of these dynamic organizational systems. These proteins represent a unique class of biological macromolecules characterized by their lack of a well-defined three-dimensional structure. They often undergo liquid-liquid phase separation (LLPS) to form membraneless organelles, also referred to as liquid droplets. Dysregulation of these liquid droplets can lead to the formation of neurotoxic oligomers and fibrils, contributing to the development of various conditions such as amyotrophic lateral sclerosis, multiple sclerosis, spinal muscular atrophy, Alzheimer’s disease, and Huntington’s disease. The aggregation propensity of IDPs is influenced by various environmental factors, making the characterization of their interaction dynamics particularly challenging. The aim of this research is to explore the conformational behavior of hnRNP A1 protein, with a focus on identifying the key factors that drive their phase separation and protein aggregation.

Methodology: X-ray scattering, specifically small-angle X-ray scattering (SAXS), was conducted to gain structural insights, focusing on the protein’s conformational dynamics and behavior under varying ionic concentrations. Utilizing supercomputers, we generated atomistic trajectories for each bead representing the proteinic structure of hnRNP A1. Through coarse-grained molecular dynamics (CG-MD), we investigated the molecular forces driving liquid-liquid phase separation (LLPS) of the prototypical IDP and explored the multi-chain association processes across different length and time scales. The expected experimental results would serve as benchmarks to validate our simulations, which utilize the explicit solvent Martini 3 force field and its recent GO model implementation.

Results: Our simulations revealed that increasing ionic concentrations led to conformational changes in the intrinsically disordered region (IDR) of the protein, resulting in a more compact structure, indicative of phase separation from the cellular environment. Not only did our findings align with laboratory data regarding protein dynamics, but we also identified limitations in the Martini 3 Go force field, suggesting potential areas for refinement in the model.

Conclusion & future research: Our study revealed that increasing ionic concentrations cause significant conformational changes in the intrinsically disordered region of hnRNP A1, driving phase separation from the cellular environment. Looking ahead, molecular dynamics simulations hold immense potential to drive breakthroughs in understanding molecular interactions. By simulating complex systems, we can explore not only the behavior of proteins but also the impact of small molecules, such as drugs, on phase separation. This approach could open new doors to therapeutic innovation by providing a deeper understanding of how molecular interactions influence disease mechanisms, offering promising pathways for drug development targeting neurodegenerative diseases.

Keywords: Intrinsically Disordered Proteins (IDPs), Liquid-Liquid Phase Separation (LLPS), Molecular Dynamics Simulations, Aggregation Propensity.

Abstract

Background and aim: Prenatal diagnosis enables the diagnosis of a wide spectrum of genetic abnormalities, mutations, genetic diseases related to the X chromosome, neural tube defects, before the birth of the fetus. Invasive prenatal procedures include amniocentesis, chorionic villus sampling, cordocentesis. Amniocentesis is currently the "gold standard" test for obtaining fetal DNA and diagnosing fetal aneuploidies. In this cohort study we evaluate and analyze the trends of aneuploidies  and the impact of maternal age on the clinical manifestation of trizomies 13,18 and 21. 

Methodology:  The criteria for their application are evidenced, and the epidemiological data collected during years 2011-2024 in the Queen Geraldine Hospital are analyzed. The molecular techniques used for the detection of aneuploidies are MLPA and QF-PCR, targeted different regions within chromosomes 13, 18, 21, X and Y as well as 15, 16, 22 only in product of conceptions.  We processed and analyzed the data with SPSS version of year 2020 for correlation between different factors as maternal age and aneuploidy; trisomies and biochemical or morphological examination suggestions among the 101 samples submitted for molecular analysis in the laboratory during this period. 

Results: To the all samples analyzed, the overall frequencies of trisomy  21 was 8.9% and  the main incidence ratio 1:300 of the newborns; trisomy 18 was 2.97%  and the main incidence ratio  1:1100; trisomy 13 was 0.99% and the main  incidence ratio   1:3000 of the newborns.  The was an increasing trend of trisomy 21during these 4 years of study.  There was a significant relationship between trisomies and spontaneous miscarriages [r= .242*, p< .015]. There was a positive relationship between aneuploidies and maternal age [r= .131*, p< .001.

Conclusions and future research: Chromosomal anomalies in general and trisomies specifically show an increasing trend through years. According to this study trisomies were the main cause of recurrent pregnancies loss and the maternal age has significant impact. It will be recommended that more researches with chromosomal anomalies on focus  must be realized because it is observed an increasing trend last decade wich can have different causes like molecular, genetic, familiar prognosis and environmental that should be defined and analyzed. 

Keywords: Aneuploidy, spontaneous abortion, trisomy, MLPA, QF-PCR.

Abstract

Background and aim: Excipients are all ingredients in a pharmaceutical formulation other than the active pharmaceutical ingredient (API). Even though excipients are traditionally considered "inactive" substances, these components can occasionally have unintended pharmacological effects. Recent studies have shown that some commonly used excipients in pediatric formulations are unsuitable for children. Substances such as propylene glycol, ethanol, and parabens, are known to cause a range of harmful events, including gastrointestinal disturbances, hypersensitivity reactions, and neurotoxicity, particularly in younger age groups of the pediatric population. Databases on the safety and toxicity of inactive ingredients have prioritized these excipients, indicating the critical need to evaluate their safety when included in children’s medicines. This study aimed to identify excipients of concern for the pediatric population, assess the extent of exposure to these potentially harmful substances in commercially available oral pediatric medicines, and evaluate the associated health risks.

Methodology: A comprehensive review of existing literature and safety databases was done to identify potentially harmful excipients for the pediatric population. A list of oral liquid formulations for pediatric use was compiled using the drug register published by the National Agency for Drugs and Medical Equipment in Albania. For each formulation, the Summary of Product Characteristics (SmPC) was examined to detect the presence of potentially harmful excipients for the pediatric population. 

Results: Following the literature review, ten excipients—propylene glycol, ethanol, polysorbate 80, benzyl alcohol, parabens (including propyl and methyl), aspartame, sorbitol, benzoic acid, and sodium benzoate—have been identified as potentially harmful for the pediatric population, especially for neonates, infants, and toddlers. A total of 257 oral liquid formulations, both ready-to-use and those requiring reconstitution, were included in the investigation. All the formulations examined for potentially harmful excipients were authorized for at least one pediatric age group. More than half of the formulations analyzed were classified as non-prescription medicines. The excipient composition of the pediatric formulations varied, with a median of two potentially harmful excipients per formulation and a maximum number of six. Among the potentially harmful excipients, sorbitol was the most frequently identified excipient in pediatric formulations (n=87), followed by methyl-paraben (n=80) and sodium benzoate (n=73). The highest number of excipients of concern was identified in ready-to-use liquid formulations.

Conclusion & future research: This study shows the presence of potentially harmful excipients in commercially available oral pediatric medicines in Albania. Multiple excipients of concern in pediatric formulations raise the possibility of cumulative exposure in children. Future research should prioritize investigating the long-term health impacts of commonly used excipients in pediatric medicines and developing comprehensive recommendations to minimize exposure in vulnerable populations.

Keywords: Pediatric, excipients, liquid, formulations.

Abstract

Background and aim: The spread of antimicrobial resistance (AMR) is a multidimensional problem which poses a threat for the public health and also affects environment. Antibiotic resistance occurs when bacteria survive in the presence of antibiotics that once effectively inhibited or killed them. Antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARG) can enter the environment from sewage sources such as hospital and municipal wastewaters and also from veterinary and agricultural practices. AMR monitoring of water environment such as surface water and wastewater has been widely applied to help in mitigating the spread of antibiotic resistance through waterborne sources. The aim of this study is to discuss the importance and some findings of antibiotic resistance monitoring in water environments using qPCR as a surveillance tool.

Methodology: For this purpose, scientific articles and recent literature reviews related to antibiotic resistance monitoring in water environments were searched. AMR monitoring of water environment can assess the circulating ARGs, identify potential hot spots for dissemination of antibiotic resistance and inform human health risks. 

Results: Gene targets that have been commonly measured by qPCR include ARGs associated with anthropogenic sources of antibiotic resistance such as Sul1 and Intl1. Also ARGs of clinical concern such as ARG encoding for extended-spectrum β-lactamase have been often monitored with qPCR in water environments and have been mostly detected in wastewater sources. 

Conclusion: Taking in consideration benefits and drawbacks,  qPCR is a sensitive method for surveillance and has been widely applied for quantifying different ARGs in a microbial community from water environments. 

Keywords: antibiotic resistance monitoring, antibiotic resistant gene , qPCR, wastewater, surface water

Abstract

Background and aim: Over millennia, humans have used plants for medicinal purposes. This article will introduce medicinal plants' knowledge and uses in Albania. 

Methodology: In this study, we were based on an article by Bajrami 2023 on the status of ethnobotany in Albania. A total of 16 ethnobotanical articles were cited in the study. For the scope of this review, 12 articles were selected based on fieldwork criteria and protocol used for conducting ethnobotanical research.

Results: 226 botanical taxa were reported for use in folk medicine, falling into 77 families. The most common families reported were Asteraceae (10%), Rosaceae (9%) and Lamiaceae (8%). Therapeutic applications of medicinal plants mostly were related to treating digestive, skin-ear-eyes-hair wounds, respiratory, and genitourinary system disorders. From the data collected, it was observed that people used mostly fruits (23%), followed by aerial parts (21%), leaves (21%), flowers (12%), and seeds (7%) with several administrations described. Additionally, 47 medicinal plant species have been recorded to have more than three medicinal properties. The most cited botanical taxa considered to have more than seven different medical properties were Urtica dioica L., Cornus mas L., Rosa canina L., and Hypericum perforatum L. 

Conclusions: This study provides a first-time ethnobotanical data analysis in Albania and offers a deeper understanding of people's use of plants for medicinal purposes. Additionally, these data could have implications for plant conservation and biodiversity in Albania and future pharmacological studies. 

Keywords: ethnobotany, medicinal plants, health, Albania, Balkan region 

Abstract

The Vjosa basin shelters a high diversity of aquatic and terrestrial ecosystems, which host rare and endangered habitats and species. Since 10 years ago, the Vjosa/Aoos River has been identified as the last free-flowing river in Europe, outside of Russia. In March 2023, the Albanian government declared Vjosa and its tributaries, Drino, Bença and Shushica as a National Park (IId IUCN Category). Among the living river organisms, algae, mainly those attached to submerged surfaces (periphyton), constitute important groups of aquatic biodiversity and productivity. They are primary producers, capable of producing organic compounds (i.e. sugars) from light energy through photosynthesis, and producing also O2, using inorganic salts, water and CO2. Moreover, they offer many other ecosystem services.  

About 50 periphyton samples were collected in representative stations of Vjosa, Drino, Shushica, Bença, Kardhiqi, Zagoria, Langarica, Sarandaporo, etc., during the years 2022-2024. Samples were observed either fresh or prepared permanent slides for diatoms. Light microscopes were used: Leica DM4B & Motic BA310, with objectives up to 100x, or/and stereomicroscope Leica M205C, with magnification up to 150x, all equipped with digital cameras. For the samples collected during 2022, the Pollution Sensitivity Index (IPS; Coste 1982) was calculated and the ecological state of the related habitats was assessed.

Here we will report briefly about species according to the main algal groups found and photographed, such as charophytes (Charophyta), green algae (Chlorophyta), siliceous algae or diatoms (Bacillariophyta), red algae (Rhodophyta), algae blue-green or cyanobacteria (Cyanobacteria), etc. 287 species/subspecies were found: 234 diatoms (227 pennatae and 7 centrics), and 51 other species (13 charophytes, 20 chlorophytes, 10 cyanobacteria, etc.); the latter (non diatoms) are given for the first time for the Vjosa basin, but also for Albania. Considering even previous studies, the total number of algal species for the Vjosa basin reaches at 451 species.

The presence of some rare species was evidenced, such as: Amphora sancti-naumii, found in Sajmola (Kelcyre) and Tre Urat, Vjosa; Gomphonella ohridana relatively well present in the Cold Water spring, Tepelena, as well as in the Brataj spring; Sellaphora krsticii relatively abundant in Gjormi, Shushica; etc. The filamentous chlorophyte Cladophora glomerata was widespread and relatively abundant in Vjosa. Charophyte filamentous Spirogyra species, as well as cyanobacteria of Oscillatoria and Merismopedia were abundant in Drino, Shushica and elsewhere. Xanthophyte Tribonema species were abundant in Kardhiqi river. Filamentous red algae Bangia atropurpurea, considered rare and threatened in many European countries, was found abundant in the river Bença.

The IPS values for 22 periphyton samples were almost high: belonging to the ecological quality 'Good' or 'Very good'. However, the summer blooms of filamentous algae of Cladophora, Spirogyra, Oscillatoria, etc. were observed; some of them, known as nitrophilous algae, thrives where the nitrogen content is high; it evidence the impact of untreated water of urban origin, but also from agriculture, livestock or other services in the area. Oscillatoria species or other cyanobacteria are known to be toxic to the aquatic biota and to humans. The knowledge of algae is important to complete better the aquatic biodiversity, the ecosystem health and other services, as well as to assess the ecological state of related aquatic habitats.

Keywords: Vjosa/Aoos NP, non vascular plants, river aquatic biodiversity, river ecological state.

Abstract

Captivity in settings such as zoos, aquariums, and research facilities has been shown to cause significant health issues, including obesity, deformities, and diseases resulting from limited space, poor diet, and lack of exercise. Physiologically, animals in captivity often suffer from chronic stress, and psychologically, many species exhibit distress in the form of anxiety, depression, and stereotypic behaviors, which are indicators of poor mental health and environmental deprivation. Case studies on species such as elephants, bears, and primates illustrate how confinement disrupts natural behaviors and social structures, leading to unnatural aggression. This literature review explores the wide-ranging impacts of captivity on animal welfare, covering physical, psychological, social, and ethical dimensions.

This review employs a systematic methodology, synthesizing findings from peer-reviewed articles, books, and case studies published over the past two decades to identify recurring patterns and gaps in the literature. Ethical concerns are also examined, particularly regarding the morality of keeping animals in confined spaces for human entertainment or research purposes. 

The results demonstrate a consistent trend of negative welfare impacts, including chronic stress and behavioral disorders, while also underscoring the potential of alternatives such as sanctuaries and wildlife reserves to provide more naturalistic and humane environments. These findings aim to offer insights into improving animal welfare and guide future research on sustainable alternatives to traditional captivity models.

Keywords: animal welfare,captivity, physiological effects, physical health, ethics

Abstract

The Eurasian Thick-knee (Burhinus oedicnemus) is a nocturnal, ground-nesting bird species distributed across southern Europe, North Africa, and parts of Asia. It typically inhabits dry, open landscapes, such as grasslands, heathlands, and agricultural areas. In Albania, this species occurs only during the breeding season and migrates south to Africa in winter. It breeds almost all along the coastal wetlands and some of the southern rivers of Albania. Unlike many ground-nesting birds that form colonies, this species is largely territorial during the breeding season. Breeding pairs establish and defend territories from other pairs, ensuring exclusive access to nesting sites and nearby feeding areas. Territory sizes can vary depending on habitat quality and resource availability. In this study, we aimed to measure the breeding territory size of this species in Narta Lagoon using an innovative technique: GPS tracking. A bird in the nest was caught in early July 2024 and equipped with a satellite transmitter. The tagged bird was tracked throughout the entire incubation period and early chick-rearing phase. Results show that the tagged bird moved across a total area of 27 ha around the nest during the tracking period. Nevertheless, 95% of the time, the bird remained within a rectangle-shaped area of 5 ha around the nest, indicating this as the size of the breeding territory for this specific individual. It is important to note that these results are specific to the single tagged individual and not representative of the entire population of Eurasian Thick-knees in Narta Lagoon. While the findings provide valuable insight into the spatial behavior of this individual, further research with larger sample size and additional seasons is necessary to validate these results and assess whether they apply more broadly across the population. This study serves as an important first step, highlighting the need for expanded efforts to understand the breeding ecology of this species better.

Keywords: Eurasian Thick-knee, Breeding territory size, Narta Lagoon, Albania

*These two authors have equal contributions 

Abstract

Sea ice is a critical indicator of climate change, playing an essential role in regulating the Earth’s energy balance, driving ocean circulation, and supporting ecosystems. Over the past few decades, rapid and unprecedented changes in sea ice extent have raised concerns about their environmental, societal, and global impacts. This study examines the long-term trends, seasonal patterns, and interhemispheric differences in sea ice dynamics using data from the National Snow and Ice Data Center (NSIDC) spanning 1978 to 2015.

The analysis was carried out using R, a statistical programming language known for its robust data analysis and visualization capabilities. The dataset was preprocessed to handle missing values and ensure data integrity. Descriptive statistics and graphical methods were employed to explore overall patterns, while time-series analysis and seasonal decomposition were used to identify trends and fluctuations. Comparisons between Arctic and Antarctic sea ice dynamics provided insights into their contrasting behaviors.

Results indicate a dramatic decline in Arctic sea ice extent, particularly during the summer months, consistent with the impacts of global warming. Conversely, Antarctic sea ice exhibited more complex and erratic behavior, marked by periods of modest increases followed by sharp declines. Seasonal decomposition revealed distinct variability in both hemispheres, with anomalies often tied to regional climatic conditions.

These findings highlight the asymmetrical responses of Arctic and Antarctic sea ice to climate change, driven by both regional and global factors. The study underscores the importance of urgent mitigation strategies to slow ongoing sea ice loss and its far-reaching consequences on ecosystems, weather patterns, and global systems. Future research should incorporate more climate variables, develop advanced predictive models, and further explore the broader implications of sea ice dynamics on planetary systems.

Keywords: sea ice extent, temperature variability, climate change, Arctic, Antarctic, statistical analysis

Abstract

17 essential chemical elements play a direct role in the plant’s metabolism; without any of them, the plant cannot complete basic functions or complete its life cycle: macronutrients: Carbon, Oxygen, Hydrogen, Nitrogen, Potassium, Calcium, Phosphorus, Magnesium, Sulphur, Iron, Chlorine; micronutrients: Manganese, Zinc, Boron, Copper, Nickel, Molybdenum; another cannot substitute deficiency of each of them. Carbon and oxygen are provided mainly from the air via leaves (absorbing CO2), oxygen and hydrogen by H2O and all the other essential elements absorbed from soil solution in the form of ions by the young part of the root (NO3-, NH4+, K+, Ca2+, Mg2+, H2PO4-, PO42-, SO42-, SiO2, Cl-, Fe3+, Fe2+, H3BO3, Mn2+, Na+, Zn2+, Cu+, Cu2+, Ni2+, MoO42-). It is called mineral salt absorption, and normally it takes place using metabolic energy. Often it is in symbiotic association with fungi known as mycorrhizae. Therefore, plant nutrition is important to nature and also of great interest to humans. Much progress has been achieved in the growth of agriculture worldwide and in Albania over the past century. This can be shown in the increase of species of economic interest, in the area expansion of the different agricultural crops, in the increase of their diversity, as well as in the continuous increase of yields and their market quality.

This study examines how plant nutrition is taught in pre-university curricula across Albania and Kosovo. We analyze the treatment of plant biology, particularly mineral nutrition, across different educational levels in both countries. Our focus extends beyond scientific content to evaluate teaching methods and practical applications, whether in classrooms, school gardens, or local farms. The methodology was based on a careful examination of the existing teaching plans of Science in pre-university education in Albania, hosted by the Quality Assurance Agency of Pre-university Education (IZHA), and in the same curriculum in Kosovo hosted by the Ministry of Education, Science, Technology and Innovation (MAShT). To update and broaden the knowledge and best teaching practices on plant nutrition, we used the existing literature in Albanian and foreign languages, and the relevant information in the biology textbooks of the pre-university system.

We note that between the two curricula (Albanian and Kosovo) there are significant differences in hours in plant biology topics (t = 1.3326; df = 22) in all three levels of pre-university education (Elementary, Lower & High Secondary Education). Moreover, plant nutrition is more clearly expressed in the Albanian Curriculum, at all levels. In the subject Knowledge of Nature (Dituri Natyre) in Albania, attention to plant biology issues can be considered satisfactory (up to 53%), while it drops down in the two above levels (up to 15% in the High School). The same approach is also observed in Kosovo but with significantly less hours than the Albanian curriculum (from 23% to 13%).

Ours are simply some empirical observations, as we have not considered too much into the content of plant biology topics in both curricula and at each level. However, we recommend the relevant experts consider the better unification of the teaching plans in both curricula showing more attention to the plant biology issues, including those related to the plant's nutrition. Moreover, we advise proper practical and environmental education to practice more teaching techniques and methods that encourage teaching and learning from practical aspects of plant biology, including plant nutrition, but not only.

Keywords: Albania & Kosovo, Pre-university Education, Biology subject, plant mineral nutrition, teaching techniques.

Abstract

Background and aim: Infections may play a role in dementia pathogenesis. Prior studies on the relationship between Helicobacter pylori (HP) infection and Alzheimer's disease (AD) have reported contradictory findings. We investigated whether HP serology is associated with an increased risk of cognitive outcomes, including dementia, Cognitive Impairment (CI), AD, and dementia-free survival (DFS) among participants (n=1382) in the Trøndelag Health Study (HUNT). 

Methodology: Serum HP antibody titers (n=5180) were assessed randomly using enzyme immunoassays, Pyloriset EIA-IgG. 22 years later, cognitive assessments were conducted using standardized cognitive assessments, DSM-5 criteria, and proxy interviews. Multiple logistic regression models (n=1337) adjusted for sex, age, and education were used to estimate odds ratios (95% CIs) for the cognitive outcomes. These were stratified by sex, age, education, C-reactive protein (CRP) levels, and APOE-ε4 carrier status and further adjusted for lifestyle, and co-morbidity risk factors during sensitivity analysis. Cox regression models were used to estimate hazard ratios (95% CIs) for all-cause mortality.

Results: In this population-based cohort study, HP titers and seropositivity (≥ 300 titers), were not associated with dementia, CI, AD, or DFS. Sub-group analysis showed no associations between HP and the cognitive outcomes among females, older participants, higher CRP levels, and APOE-ε4 carriers. No associations were found after further adjustments by co-morbidities and lifestyle confounders.  

Conclusion & future research: HP was not associated with later dementia or AD risks in the HUNT study. The relationship between infections and neurodegenerative diseases should be explored further.

Keywords: H. Pylori, Infection, Dementia, Cognitive Impairment, HUNT

Abstract

Background & aim: Pheochromocytoma is a rare tumor that develops in the adrenal medulla, the inner part of the adrenal glands. This tumor leads to the excessive release of catecholamines, resulting in symptoms such as rapid heart rate, high blood pressure, anxiety, intense headaches, excessive sweating, and unintentional weight loss.

Methodology: This study studied 51 suspected patients, and 17 of them had pheochromocytoma. Levels of catecholamines and their byproducts in the blood and urine are measured. The analysis includes measuring plasma-free metanephrines or fractionated metanephrines in urine. Additional tests, such as measuring total urinary metanephrines, plasma or urinary catecholamines, or urinary vanillylmandelic acid (VMA), will be used to confirm the presence of the tumor. 

Results:  We detect elevated levels of catecholamines or their byproducts in the blood and urine of patients with pheochromocytoma . The most accurate tests include measuring plasma-free metanephrines or fractionated metanephrines in urine. Additional tests, such as measuring total urinary metanephrines, plasma or urinary catecholamines, or urinary vanillylmandelic acid (VMA), will be used in future steps to confirm the presence of the tumor.

Conclusion & Future Research: This study highlights the diagnostic value of plasma-free metanephrines and fractionated urinary metanephrines as the most accurate tests for detecting the tumor. Elevated levels of catecholamines and their byproducts provide critical evidence for diagnosis, while additional confirmatory tests, including urinary vanillylmandelic acid and total catecholamine measurements, can strengthen diagnostic accuracy. Early and precise diagnosis is essential to prevent life-threatening complications such as hypertensive crises, arrhythmias, or stroke. Future research should focus on refining diagnostic algorithms, incorporating novel biomarkers.

Keywords: Adrenal Tumors, Catecholamines, Metadrenalines, Phaeochromocytoma, Hypertension

Abstract

Background & aim: The pursuit of an ideal body image is a significant psychological and sociocultural issue that often leads individuals to neglect their overall health. This study aims to investigate how much individuals are willing to compromise their health in pursuit of a perfect body image. It seeks to examine the long-term health effects of this behavior and to determine whether individuals understand the importance of nutrition for their overall well-being. 

Methodology: This study utilizes a survey done on 439 individuals, of various nationalities and demographic zones, to assess the exploration of social media usage, body image perceptions, and dietary choices among participants of diverse age groups, from 13 years old to older than 65 years of age. We looked at their behavioral patterns related to their usage of social media and choices of nutrition, which led us to learn more about their perception and attitudes regarding the influence that social media has on their lives and the effects on their health

Results: After conducting thorough research and analyzing survey data on software such as R and Excel, we found that social media has slightly influenced 41.8% of our sample to change their eating habits or feel pressure to do so. Additionally, 74.7% of respondents believe that the reliability of nutrition advice posted on social media can vary, while 50.3% think that following a restrictive diet is a good idea for weight loss. When asked about their emotions surrounding food, 29.1% of the sample reported that sometimes they feel guilty after consuming foods high in carbohydrates. Many expressed the opinion that social media creates the impression that the only way to achieve a slim and healthy body is to restrict food intake. Finally, 46.8% of our sample believe that striving for an ideal body image can slightly negatively impact both quality of life and health and 25.1% think that this impact is more serious.

Conclusion: The study concludes that social media influences dietary behaviors and perceptions of body image, often leading individuals to adopt restrictive eating patterns and experience negative emotional and health impacts.

Keywords: Body Image, Long-term health effects, Nutrition, Perception, Eating habits, Psychological causes and effects.

Abstract

Background and aim: Epstein–Barr Virus (EBV) otherwise known as human herpesvirus 4, is one of eight known human herpesviruses. It infects more than 95% of the world's population, especially adolescents and young adults. Children are usually asymptomatic or moderately ill. This infection occurs with an increased of C-Reactive Protein (CRP), lymphocytes, and the production of IgM and IgG antibodies against EBV in the human body. This study aims to identify the incidence and correlation between EBV antibodies titer, CRP concentration, and Whole Blood Cells (lymphocytes and monocytes). 

Methodology: We collected human blood samples from patients presenting with symptoms suggestive of viral infection. The samples were processed to test for the presence of EBV antibodies (IgG and IgM). C-Reactive Protein levels were quantified using high-sensitivity CRP assays to assess systemic inflammation. Complete Blood Count (CBC) tests were performed to measure whole blood parameters, focusing on lymphocyte and monocyte counts. All procedures followed standard clinical laboratory protocols. Demographic data, including age and gender, were recorded for each participant to analyze trends across subgroups. The study adhered to ethical guidelines, with informed consent obtained from all participants or their guardians. Data were analyzed to determine correlations between EBV antibody titers, CRP concentrations, and variations in white blood cell counts.

Results: From a total of 666 patients, 206 of them were tested for CRP, EBV IgG/M and WBC. 59% of the group were males (mean age 14 years) and 41% were females (mean age 16 years). Overall, 86% of our group were positive about the presence of EBV IgG antibodies in serum. It was observed that 35 positive IgM samples had an increase number of lymphocyte cells while only 13 of them had an increase of CRP. 

Conclusion & future research: Such tests help physicians to better understand the clinical situation of the patient by correlating EBV antibody titers, CRP concentration, and changes in blood cell counts. This correlation aids in the accurate diagnosis and monitoring of EBV infections, particularly in adolescents and young adults where symptoms may vary. Future research should focus on longitudinal studies to explore the progression of EBV infections and their immunological impacts over time. Additionally, investigating the role of other inflammatory markers and their relationship with EBV antibody titers could provide deeper insights into the systemic effects of the virus. Expanding the sample size and including diverse populations may also enhance the generalizability of the findings. 

Keywords: viral infections, EBV antibodies, CRP, whole blood cells

Abstract

Background and aim: Glioblastoma is one of the most aggressive and heterogeneous brain tumors, with varying cell subpopulations that complicate both diagnosis and treatment. Understanding the gene expression profiles of individual cells can help identify distinct cell subtypes, thereby improving therapeutic strategies. Single-cell RNA sequencing (scRNA-seq) offers a powerful tool to study this heterogeneity, but the high dimensionality of scRNA-seq data poses challenges in identifying relevant biological patterns. This study aims to evaluate the effectiveness of matrix factorization techniques, including Principal Component Analysis (PCA), Non-Negative Matrix Factorization (NMF), and Bayesian Non-Negative Matrix Factorization (Bayesian NMF), in identifying glioblastoma cell subgroups. 

Methodology: The study uses publicly available single-cell RNA sequencing data from glioblastoma patients, obtained from http://www.gbmseq.org/. The data comprised 3,589 cells sampled from both the tumor core and surrounding peritumoral tissue. Before applying statistical analysis methods, the data underwent preprocessing following the steps below: Quality control, consisting of removing cells with a high proportion of mitochondrial gene expression and retaining genes expressed in at least three cells to minimize noise from rare or low-quality transcripts; Normalization of the data; Feature selection, identifying the most highly variable genes to focus on features that contribute the most. Three dimensionality reduction methods were applied: PCA, to reduce dimensionality and capture key variation in gene expression(helps in highlighting differences between cell types); NMF to decompose the gene expression matrix into non-negative factors, highlighting underlying patterns and identifying specific cell subtypes; Bayesian NMF to perform matrix factorization with uncertainty justification, providing probabilistic insights into cell grouping, especially detecting infiltrating neoplastic cells. Visualization techniques were used to compare cell clusters and evaluate the ability of each method to identify biological patterns relevant to glioblastoma heterogeneity.

Results: We visualize the results of each method and evaluate their performance based on computational efficiency (execution time) and biological accuracy (clustering quality). Comparison of visualizations from each method highlights PCA’s broad clustering, NMF’s detailed structure, and Bayesian NMF’s confident separation of cell subpopulations. Differences between core and peritumoral cell profiles suggest unique gene expression trends, potentially reflecting the tumor microenvironment. In the tumor core, cells expressed genes involved in regulating cell size to adapt to hypoxia, meeting energy demands for proliferation and activating survival pathways. In peritumoral cells, genes supporting cell-matrix interactions, mitochondrial function, migration, and oncogenic drivers were prevalent, promoting invasion in the periphery. 

Conclusion & future research: The preliminary explorations recommend Bayesian NMF for capturing glioblastoma heterogeneity with uncertainty estimates, while PCA and NMF are noted for exploratory analysis and identifying distinct biological patterns. The work is ongoing and will further compare these methods using the NMI criterion. 

Keywords: Glioblastoma, Cell heterogeneity, Matrix factorization, Dimensionality reduction, scRNA-seq

Abstract
Background and aim: Cortisol, often referred to as the "stress hormone," is essential for stress response and homeostasis. It is involved in a wide range of physiological processes, including metabolism, immunity, and inflammation regulation. Chronic cortisol dysregulation levels have been associated with various metabolic disorders, such as obesity, insulin resistance, and cardiovascular disease. This review aims to explore the complex role of cortisol in stress response and metabolic disorders.

Methodology: A systematic review of the literature was conducted, with studies published between 2000 and 2024 in databases such as PubMed and Google Scholar. The review targeted studies exploring the relationship between cortisol, metabolic disorders, and chronic stress. 

Results: Studies show a strong correlation between elevated cortisol levels and the development of metabolic disorders, especially in individuals exposed to chronic stress. Continuous high cortisol levels were associated with increased visceral fat accumulation, insulin resistance, abdominal obesity, and elevated blood pressure, all of which together elevate the risk for metabolic syndrome and cardiovascular disease. The risk of developing metabolic syndrome was reported to be up to 30-50% whereas the risk of developing a cardiovascular disease was about 20-40%. Individuals with morning cortisol levels higher than 500 nmol/L are were reported to be more likely to have higher amounts of visceral fat and a higher risk of developing type 2 diabetes. Cortisol is important for short-term stress adaptation, but its long-term dysregulation seems to be a problem for metabolic health.

Conclusion & future research: Cortisol plays an important role in mediating stress response, but its chronic overproduction may contribute to the development of metabolic disorders. Future investigations should focus on the impact of lifestyle interventions, such as diet and exercise, on cortisol regulation.

Keywords: cortisol, stress, metabolic disorders, insulin resistance, cardiovascular disease 

Abstract

Background and aim: Alzheimer’s Disease (AD) is a progressive neurodegenerative disease in which nerve cells in the brain die. AD is the most common cause of dementia, accounting for almost two-thirds of dementia cases, as it primarily affects the hippocampus and entorhinal cortex. AD is linked to other diseases such as cardiovascular diseases and diabetes, which worsens the quality of life of the individual affected. AD is characterized by the accumulation of both amyloid β plaques and the accumulation of phosphorylated tau proteins. These amyloid β plaques have a high affinity to the receptors of the nerve cells, causing the synapses they form between nerve cells to erode, therefore inhibiting the communication between neurons and leading to the cell their death. DNA methylation, an epigenetic mechanism that is involved in the expression of genes primarily regulated by DNA methyltransferase (DNMT), is involved in the reason why we have an overproduction of amyloid β. DNA methylation is primarily regulated by DNA methyltransferase (DNMT).  This review aims to understand the epigenetic mechanism of the amyloid β plaque in Alzheimer’s disease. 

Methodology: This review study is based on articles retrieved from PubMed and JSTOR databases. The search engine Google Scholar was also used to search for said articles. The articles were searched by using the following keywords: “Alzheimer’s disease”, “DNA methylation”, “Amyloid β plaques”, “DNMT”, “Neurodegeneration”, and “Epidemiology of AD”. The selection of the papers was done by reviewing their abstracts and assessing their relevance to the topic, then the whole paper was evaluated; those who did not have any correlation to the topic were excluded. Only English-based papers were reviewed.  

Results: Amyloid beta precursor protein (APP), Beta-Site Amyloid Precursor Protein Cleaving Enzyme (BACE), Presenilin 1 (PS1), and Presenilin 2 (PS2) were found to be the most relevant genes related to the formation of amyloid β plaques. The modification in expression levels of these genes was related to AD.  Genetic studies performed in AD patients showed that the promoter regions of the APP gene were found to be hypomethylated, which increased amyloid β production. In addition, the increased expression of APP, and induction of hypomethylation of PS1 gene promoters increased amyloid β plaques in the BV2 (mouse microglial cells). Similar findings were also found for the BACE and PS2 genes concerning the production of the amyloid β plaques.  It was also observed that in AD and other neurodegenerative patients, there were dysregulations with DNMT. 

Conclusion & future research: The understanding of the epigenetic mechanism in the formation of amyloid β plaques is crucial, which would potentially help in developing therapeutic strategies to prevent or reverse plaque formation in AD patients. Further research needs to be conducted to see the effect on DNMT-inducing drugs. that are specific to the amyloid β protein production and how they could be used to treat Alzheimer’s disease. 

Keywords: Alzheimer’s Disease, Epigenetics, DNMT, Amyloid β plaques

Abstract

Background and aim: Zinc oxide nanoparticles (ZnONPs) have gained significant interest for their potential applications in the field of medicine. Due to their unique morphology and high reactivity, ZnO-NPs hold considerable promise in various biomedical applications such as drug delivery, antibacterial and antifungal activity, wound healing, anticancer applications, and anti-inflammatory effects. They have shown potential interactions with blood cells, particularly erythrocytes, raising concerns about potential adverse effects. Thus, studying the cytotoxic effects of ZnONPs on erythrocytes provides insights into their potential impact on human health. This study delves into the intricate interplay between ZnONPs and human erythrocytes, aiming to provide a comprehensive understanding of the mechanisms governing red blood cell toxicity response. 

Methodology: Blood samples were drawn from healthy individuals of our staff after taking their consent according to the Albanian Law and Ethics Committee. The blood was taken in EDTA tubes and processed within an hour. For morphologic examination the blood was centrifugated for 10 min at 3000 rpm in order to divide the plasma from the cells. The plasma was removed and 1 ml of erythrocyte volume was taken and diluted in 3 ml of physiologic solution (NaCl 0,9%) to wash them from remaining plasma and protein traces. The procedure was repeated 3 times until a clear supernatant was acquired. Then 1 ml of the erythrocyte mass was put in 4 different tubes and exposed and incubated at different concentrations of ZnO-NPs (12.5, 25, 50 and 100 µg/ml) for 3 and 24 hours. The morphological alterations were determined under light microscopy. For hemolysis examination the blood as taken was divided in 4 tubes each of 1 ml and directly exposed and incubated to the different concentrations of ZnO-NPs (12.5, 25, 50 and 100 µg/ml) for 3 and 24 hours. Then each tube was centrifugated for 10 min at 3000 rpm to divide the plasma from the cells. The plasma only was taken from each tube and examined in a Sysmex XS-1000i Blood Analyser for presence of hemoglobin traces.

Results: Under light microscopy examination the presence of echinocytes (abnormal erythrocytes) was detected in each concentration and time exposure. Echinocytes were counted by eye in 10 different high fields and a concentration dependent rising number of them was observed from 10 to 70 % of the erythrocyte total number in the 3 hours exposure. Similar findings were observed in the 24 hours exposure but with limitations because of hemolytic effect. The hemolytic examination of the plasma revealed dose dependent presence of hemoglobin for the 3 and 24 hours exposure. Respectively, in 3 hours exposure the hemoglobin values were 0 g/dL, 0,2 g/dL, 0,4 g/dL and 0,6 g/dL for ZnONPs concentrations of 12,5 µg/ml, 25 µg/ml, 50 µg/ml and 100 µg/ml. The P value was statistically relevant p = 0.040833 (p < 0.05). For the 24 hours exposure the hemoglobin values were 0,2 g/dL, 0,2 g/dL, 0,4 g/dL and 0,7 g/dL for ZnONPs concentrations of 12,5 µg/ml, 25 µg/ml, 50 µg/ml and 100 µg/ml. The P value was statistically relevant p = 0.008864 (p < 0.01). 

Conclusion: Our results demonstrate that ZnONPs caused hemolytic effects through cell membrane destabilization, and hemolytic processes as primary toxicological outcomes leading so toward eryptosis. Our findings reveal dose-dependent cytotoxic effects of ZnONPs on human erythrocytes, mirroring their potential adverse effects on circulating red blood cells in vivo.

Keywords: Erythrocytes, hemolysis, eryptosis, zinc oxide nanoparticles 

Abstract

Background and aim: Tirana Lake is the most important part of the Great Tirana Park, a public park of 290 ha, built in the 1950s, which lies in the southern part of the Tirana capital. The Park and its Lake, in the southernmost part, form a green-blue (terrestrial-water) ecosystem complex, very attractive and relaxing. The Lake is an artificial basin of 48 ha, with a dam of 400 m long. The area is almost unique in its attractiveness for the whole Tirana urban area, with more than 800 thousand inhabitants, extending in more than 1,100 km2.

An algal greenish jelly layer was clearly visible on the water surface, mostly in a wide lakeshore area of the Lake during October-November 2024. We will report here briefly some data related to this phenomenon, how to deal and prevent it in the future.

Methodology: Sporadic plankton samples were collected almost weekly on 09 October near the western part of the Dam, and on 13 October, 20 October, 03 November 2024 on the eastern lakeshores. The samples were observed fresh or stained with Lugol solution in the Leica DM4B light microscope and Leica DMi8 inverted microscope, equipped with DIC optics, with Flexacam C3 digital camera, with Leica LAS X program. The determination was done using the available literature; some other experienced expert in the region was also consulted.

Results & discussion: We concluded that it was a cyanobacteria bloom of Microcystis, probably several taxa M. aeruginosa, M. flos-aquae, M. botrys. In such a dense form, it was seen for the first time by us in Lake Tirana. The blooming intensity was high during the first week of October, then vanished, and again intensified until the second week of November when the rainy and cool weather started. The bloom vanished during second-third weeks of October, probably due to weather temperature drops; the plankton there was dominated by the dinophlagellate Ceratium hirundinella, a common species in our freshwater lakes.

Microcystis blooms are formed often during warm, calm weather in lakes and ponds with relatively high nutrient content (nitrogen or phosphorus), or low nitrogen to phosphorus ratios (N:P<15). Worth mentioning that Microcystis blooms are usually toxic; but no signs of dead fish or other livings were observed in Lake Tirana. According to the assessment carried out by the Lab of the National Environment Agency, Tirana, on 16 October 2024, at the Lake Dam, the P-PO4 value was 0.055 mg/L; it means that the waters were highly eutrophic.

Conclusion & future suggestions: The algal bloom in Lake Tirana could be a probable consequence and a signal of eventual heavy nutrient load (N & P). In our opinion the nutrient origin could be probably from the leakage of the untreated urban wastewater and/or from other nearby services.

After the 1990s, urban constructions, various services, and much traffic increased around the Tirana Park, and Lake Tirana. In synergy even with climatic conditions, high temperature and heavy drop of the water level, for sure constitute a pressure on the Lake's water quality. Collecting and wastewater treatment is an urgent challenge, as is the necessity of life quality for aquatic living and humans.

Keywords: Tirana Lake, Microcystis, harmful algal blooms, wastewater treatment.

Abstract

Eurasian Thick-knee is a Palearctic bird species that occurs in parts of Europe, Asia and North Africa. The species breeds in southern and eastern Europe, including countries like Spain, France, Italy, Greece, and parts of the Balkans. Some populations are migratory and move south during winter and some are resident. Its breeding range extends into south-western Asia, particularly in regions such as Turkey, in the Middle East, and parts of Central Asia. In Africa it breeds in all the countries that are wet by the Mediterranean Sea and in Sudan. In Albania this species is a summer visitor and winters in Africa. It breeds in small numbers in the western coastal wetlands and in the river beds of the southern part of the country. From late August to mid-October, in the area of Narta Lagoon in southern Albania, pre-migration gatherings of this species regularly occur, with the peak numbers in mid-September, with up to 162 individuals counted in the 2nd week of September 2024. The aim of this abstract is to report this finding. Further research is necessary to investigate the origin of the gathered birds. 

Key words: Eurasian Thick-knee, Pre-migration gatherings, Narta Lagoon, Albania

Abstract

Background & Aim: This study presents an in-depth assessment of the ecological condition of the Zeza River (Fushë-Krujë) utilizing the Stream Visual Assessment Protocol (SVAP). The river, vital for local biodiversity and community livelihoods, has faced significant ecological degradation due to anthropogenic pressures, including pollution, habitat alteration, and land use changes. 

Methodology: By applying the SVAP Protocol, a comprehensive evaluation of various ecological indicators was conducted, including invertebrate diversity, bank stability, and aquatic habitat availability.

Results: The findings reveal critical areas of concern that contribute to the river's declining health. In response, the study proposes a series of naturalization alternatives aimed at restoring ecological integrity and enhancing the river's resilience. These alternatives encompass strategies such as riparian buffer restoration, sustainable land management practices, and community engagement initiatives to foster stewardship of the river ecosystem.

Conclusion: this research underscores the importance of adaptive management strategies and ecological restoration in maintaining the ecological health of the Zeza River. The recommendations provided herein serve as a roadmap for stakeholders and policymakers to implement effective conservation practices, ensuring the sustainable use of the river's resources for future generations.

Keywords: Protocol, restoration, management, visual, degradation.

Abstract

Background and Aim: Technology advancements are transforming biological education, and virtual laboratories are an essential part of this transformation. In this study, we aim to evaluate the integration and impact of virtual laboratories in biology curricula. The focus is on their effectiveness in improving student understanding, engagement, and overall academic performance.

Methodology: We analyzed the advantages and disadvantages of virtual laboratories in the teaching of the subject of biology, based on published scientific articles. Platforms such as Google Scholar, PubMed, ScienceDirect, and JSTOR were used to search for papers using the keywords "virtual laboratory", "virtual lab for Biology" and "Virtual lab pros and cons." Inclusion criteria focused on studies that directly explored the application of virtual labs, and student experiences, as well as the main platforms that were used to achieve the goal, such as adaptive learning platforms, AI-driven simulations, video and audio aids, etc.

Results: Studies show that virtual laboratories significantly improve students' understanding of complex biological concepts and promote active learning. Interactivity and accessibility were identified as key factors contributing to increased motivation and knowledge improvement. Educators report positive changes in teaching methodologies, with virtual labs providing flexible options for conducting experiments, especially in resource-constrained settings, they help to reduce the risk and reduce the costs of preparations and laboratory tools, and reduce the time of conducting experiments.

Conclusion & Future Research: Integrating virtual laboratories into biology education is essential for cultivating a more effective and inclusive learning environment. Future research should focus on long-term impacts on student performance, developing best practices for implementation, and exploring the potential of augmented and virtual reality technologies to further enrich biology education. 

Keywords: Virtual Laboratory, ICTs, Biological Education

Abstract 

The Little Gull (Hydrocoloeus minutus) has a broad distribution across the Palearctic region, breeding primarily in northern Europe and Asia, with the largest populations found in Russia. It also occurs in parts of North America. Its wintering grounds extend to the Atlantic coast of western Europe, the western and eastern Mediterranean Basin, the Black Sea, the Caspian Sea, and as far as the western coast of the Yellow Sea in Asia. In Albania, this species has been occasionally recorded in small numbers during winter, primarily in a few wetlands. It is also rare during migration, with only a few documented sightings. In mid-April 2024, a flock of 45 individuals was recorded in Albania. This is the first flock of over 10 individuals that it is ever recorded in Albania and the first during passage. This observation was recorded in the Important Bird and Biodiversity Area of Narta Lagoon, located in southwestern Albania near the coast. This sighting suggests that the species may be starting to use Albania's coastal wetlands more frequently during migration. However, further research is needed to confirm this assumption. The purpose of this publication is to report this notable observation.

Key words: Little Gull, Hydrocoloeus minutus, Migration, Albania 

Abstract

Background and Aim: The application of Artificial intelligence (AI) to improve teaching methods, and facilitate complex simulations of biological processes is significantly increasing. Still, there is a limited understanding of how AI is being applied in this domain and its overall effectiveness. This literature review aims to explore existing research on AI applications in biological education, identify key advancements, and evaluate the benefits, challenges, and disadvantages.

Methodology: The study systematically reviewed peer-reviewed articles, conference papers, and reports published between 2010 and 2024. Databases such as PubMed, and Google Scholar, were searched using the keywords  “artificial intelligence,” “biological education,” and “AI in teaching biology.” Inclusion criteria focused on studies that directly explored the application of AI technologies in biological education, such as adaptive learning platforms, AI-driven simulations, and intelligent tutoring systems. Articles were categorized based on their AI application type, educational level (high school, undergraduate, graduate), and reported outcomes, including student performance, engagement, and accessibility improvements.

Results: The review identified several key applications of AI in biological education. Adaptive learning platforms were shown to improve student engagement rates. Virtual labs and AI-driven simulations, particularly in subjects such as genetics and molecular biology, provide students with hands-on experience in a cost-effective manner. Despite these advancements, challenges such as the high cost of AI technology integration, the need for teacher training, and concerns over the ethical implications of AI in education were frequently highlighted.

Conclusion & Future Research: AI has shown promising potential in transforming biological education by making learning more interactive, personalized, and accessible. Successful implementation requires teacher preparedness, infrastructure, and ethical considerations. Exploring how AI can support collaborative learning and inclusivity in biological education may be a valuable direction for future studies.

Keywords: Artificial Intelligence, Biological Education, Adaptive Learning, ICTs

Abstract
Background and Aim: The integration of Information and Communication Technology (ICT) in education is essential for engaging students in today’s digital age. Traditional teaching methods often struggle to meet the needs of learners accustomed to interactive and technological solutions. This study aimed to design and evaluate a virtual biology laboratory specifically focused on the endocrine system for Higher Secondary Education (HSE).

Methodology: A systematic literature review was conducted, analyzing 15 peer-reviewed papers on the use of ICT in biology education. A focus group comprising 5 biology teachers from high schools in Tirana, Albania, provided insights into the challenges and current practices of ICT-based teaching in the region. The study involved 30 students who participated in lessons using the virtual lab. ICT tools, including Google Classroom, Google Forms, LucidChart, and Kahoot, were used to develop and deliver interactive virtual experiments. The effectiveness of the virtual lab was assessed through pre- and post-lesson questions.

Results: The systematic review highlighted the advantages of ICT in facilitating interactive and experiential learning. The focus group emphasized the need for teacher training and accessible resources to enhance ICT integration. The virtual lab allowed students to simulate hormone-related experiments and explore key endocrine processes interactively. Post-lesson quiz scores showed an improvement compared to a control group using traditional methods. Student engagement surveys revealed high levels of satisfaction with the virtual lab, particularly with its gamified and visual features.

Conclusion and Future Research: The virtual laboratory for the endocrine system significantly enhanced student understanding and engagement, as demonstrated by improved scores and positive feedback. Insights from the literature and teacher focus group underscored the importance of teacher preparedness and resource accessibility in implementing ICT solutions effectively. Future studies should explore adapting the virtual lab framework to other biology topics and assess its long-term impact on learning outcomes.

Keywords: ICT, biology education, endocrine system, virtual laboratory, student-centered teaching.