(3) Synopsis of contributions in the Life Sciences
Active member of Arts, Civic, Cultural, Educational, Music, Scientific and Squash communities.
Life Sciences:
Qualified Fundamental Principles governing Mammalian Evolution/Speciation, Chromosome/Genome Biology and Sex Determination/Gametogenesis. Either made pioneering contributions in the 6 sub-fields of Recombinant DNA/Diagnostics, DNA replication, DNA recombination, Regulation of Mammalian (Y) Evolution/Speciation, Chromosome/Genome Biology and Sex Determination/Gametogenesis or corrected or was the sole investigator to successfully reproduce literature in Cell, Science, Nature and PNAS published by 7 Research Groups at Harvard University, University of California at San Francisco, Fred Hutchinson Cancer Research Center/University of Washington, Massachusetts Institute of Technology, Jackson Labs and the Medical Research Council - London, that were at the Fore Front of these 6 sub-fields (Introduction to this site, selected Abstracts and Publications, PubMed).
Contributions in other sub-fields included Protein Export in Yeast, Structure/Function relationships of Human Cathepsin K/collagen, the Neurophysiological and Transcriptional response of the Anoxia resistant Turtle (T. scripta) Brain and Sorting and Fusion mechanisms subject to the Differential Adhesion Hypothesis (DAH) in Xenotypic and Allotypic 3D co-cultures.
Pioneered (with collaborators):
Concepts:
(1) First application of Recombinant DNA technology in the Direct Prenatal Diagnosis of a Human disease - Sickle Cell Anemia, contributing the first ever experimental results constituting the most important/relevant segments of the published results in the paper (PNAS 1981, As per: James D. Watson, John Tooze, David T. Kurtz, Pages 215, 216 & 246 in Recombinant DNA - A Short Course, 1983, 1st Edition, see link below).
(2) Systematization of the Regulation of Mammalian Chromosome/Genome Biology in Development and Homeostasis - indispensable for eventually preempting mutational causes of >6000 diseases costing tens of trillions per year as a replacement of current approaches perpetuating symptom management with therapies that are often counter-productive and yield mixed results. Furthermore, none of the mechanisms regulating Chromosome/Genome Biology that have been worked out in Mammalian Tissue Culture, Mouse Models/Gene Ablations and other Model organisms can be shown to be operational in Mammals without this Systematization and the tools that would emerge from it (1984-2012).
(3) Qualified Principles governing Mammalian Evolution, Chromosome/Genome Biology and Sex Determination. Based on (A) Epigenetic modifications of Reciprocally Backcrossed Interspecific combinations of Genomes and Y chromosomes and (B) Genetically modulated instability of the Y chromosome and the Sex Determining Sry locus showed that a re-evaluation was necessitated in mechanisms of (a) Testis Determination. Alternative explanations need to be tested. These include, (1) establishing the threshold numbers of Pre-Coelomic-Sertoli cells required for TD, as it is indispensable for, (x) determination of the minimal numbers of Sry(+) cells required for the recruitment of Sry(-) cells (and thus minimal levels of SRY) into Sertoli/Testicular Chord lineages, (y) pre-emption of Sry Intrastrand Nucleation-Branch Migration by tissue specific e.g. Neuronal Chromatin and (z) recruitment of tissue specific recombinational mechanisms as in Meiocytes, Lymphocytes and Thymocytes into DNA repair functions and therefore pre-emption of deletions of Sry in Sertoli cells, (2) single cell/molecule analyses of contributions of putative SF1-SRY-HMG-SOX9/Sry secondary structure complex formation as by-products of DNA replication to the Developmentally /Physiologically relevant regulation of Testis Determining functions of the Sry locus in Pre-coelomic and Sertoli cells of day 10 - day 12.5 Embryos. The possible contributions of these secondary structures may include maximal rates of elevation to maximal levels, followed by maximal rates of suppression that lead to the stringent Kinetic, Temporal and Spatial constraints on the transcription of Sry that are required for Testis Determination. These functions are suggested by a range of results in the literature, and even if they made relatively small contributions to Testis Determination and Development, their effects would be magnified on an Evolutionary scale by offsetting the high degree of instability and selecting for the IR structure of Sry. Such amplified Evolutionary selection and fixation of small effects of Gene structure on Expression and Phenotype have been reported with, the Ped gene regulating Blastomere number/size in mice and both rDNA magnification-reduction at the Bobbed locus and X:A ratios setting of Transcription rates of tra/tra2/Sxl and the subsequent Splicing cascades responsible for Sex Determination in Fruit Flies (Synopsis, Select Publications and Abstracts, PubMed).
Technical Innovations:
(1) Among the first successful applications of Electroblot-DBM systems for high resolution analysis of DNA (1981).
(2) Established Drift/Epigenetic Modifications of genomic Y chromosomal repeated sequences with Denaturing Formamide Gradient Gel Electrophoresis (DFGGE) (1984).
(3) Established Southern Blot coupled Nitrocellulose Filter Retention Assays (S-NFRA) for tightly/covalently bound peptides to Y elements - Epigenetic modifications (1994).
(4) Established the 'Pull Down Assay' (Streptavidin-Biotinylated DNA) for analyzing HJ-Sgs1 Helicase/Top3 Topoisomerase/Rmi1 (Yeast S.c. ortholog of Bloom's) complex formation (2005).
(5) Established a sensitive and specific Immuno-chromogenic stain for detecting exported proteins from Yeast (P.p.) (2006).
(6) First successful analyses of Sorting and Fusion interactions of Stem cells-Dermal cells and Dermal Cells-Keratinocytes in 3D Hanging Drop Cultures as per Differential Adhesion Hypothesis (2007).
Current work :
(1) Evolution, Speciation, Sex Determination.
(2) Developmentally/Physiologically Relevant Regulation of Mammalian Chromosome/Genome Biology. Genetic and Biochemical assays exploiting Evolutionary diversity of Regulatory Alleles with Interspecific Mouse Backcrosses, Y chromosomal targets, viral Origins of DNA replication in mouse cells, Holliday Junction (HJ)-DNA replication /restart (Yeast S.c. orthologs of Bloom's) complexes and Y Epigenetics.
(3) Cancer biology - the roles of the processes outline in items 2, 4, 5, 6 and 7 in Mutational Initiation, Metastasis, Drug resistance, Epigenetics, Vasculogenesis/Angiogenesis, MMP Intercellular Matrices and Protein Export/Tumor Micro-environments driving Oncogenesis.
(4) Analyses of Sorting and Fusion interactions of Stem cells-Dermal cells and Dermal Cells-Keratinocytes in 3D Hanging Drops as per the Differential Adhesion Hypothesis (DAH).
(5) Neurophysiological resistance of the Turtle brain to Anoxia and its possible role in preventing Neurodegenerative (e.g. AD) disease states. The combined application of the spectrum of Neurodegenerative diseases, Anoxia resistance, Yeast respiratory chain (rho0/petite) and protein export mutants, Mouse strains with Behavioral and Cognitive disabilities and comparative Evolutionary and Functional Genomics and Phylogeny (Yeast/Mouse/Turtle) in elucidating acquisition of Ischemic Survival, Sexual Dimorphism and Cognitive Functions by the Brain.
(6) Structure/function analysis of Human Cathepsin K - Collagen in homeostasis and in disease states.
(7) Protein export in Yeast - development of the immuno-chromogenic stain and isolation of over-secretors ( P. pastoris).