A functional model for progestogen-induced breakthrough bleeding

During the past 5 years, a number of important advances have been made in our understanding of the mechanisms of sex steroid-induced breakthrough bleeding (BTB). These observations suggest that superficial endometrial vascular fragility may be the mechanism underlying BTB, and molecular changes in the microvasculature as well as hysteroscopic observations have supported this hypothesis. This paper aims to present a unified picture of our current understanding of BTB, particularly that associated with progestogens, to indicate current gaps in our knowledge and possible directions for future research.     

Toward a genome-wide systems biology analysis of host-pathogen interactions in group A Streptococcus

Genome-wide analysis of microbial pathogens and molecular pathogenesis processes has become an area of considerable activity in the last 5 years. These studies have been made possible by several advances, including completion of the human genome sequence, publication of genome sequences for many human pathogens, development of microarray technology and high-throughput proteomics, and maturation of bioinformatics. Despite these advances, relatively little effort has been expended in the bacterial pathogenesis arena to develop and use integrated research platforms in a systems biology approach to enhance our understanding of disease processes. This review discusses progress made in exploiting an integrated genome-wide research platform to gain new knowledge about how the human bacterial pathogen group A Streptococcus causes disease. Results of these studies have provided many new avenues for basic pathogenesis research and translational research focused on development of an efficacious human vaccine and novel therapeutics. One goal in summarizing this line of study is to bring exciting new findings to the attention of the investigative pathology community. In addition, we hope the review will stimulate investigators to consider using analogous approaches for analysis of the molecular pathogenesis of other microbes.     

Inside the mammalian telomere interactome: regulation and regulatory activities of telomeres

Work in model organisms, such as mouse, yeast, Tetrahymena, ciliates, and plants, has led to a deeper understanding of telomere biology. Telomeres together with telomere-binding proteins have evolved to protect chromosomal ends and maintain chromosomal length and integrity. Over the last two decades, biochemical, molecular, cellular, and genetic studies have greatly enhanced our knowledge of the unique function and structure of telomeres and telomere-associated factors. In this review, we focus on the important advances, in terms of our knowledge and the methods used, in understanding mammalian telomere regulation by telomeric proteins. Recently, the 6 telomeric proteins (TRF1, TRF2, POT1, TIN2, RAP1, and TPP1) were found to form a high-order complex. This complex and its associated partners provide the basis for constructing an interaction map of telomere regulators in mammalian cells, which we named the Telomere Interactome. The Telomere Interactome incorporates the various telomere signaling pathways and represents the molecular machinery that regulates mammalian telomeres. The establishment of the Telomere Interactome will also enable the integration of the intricate circuitries that regulate telomeres with other cellular interactomes in vertebrates.     

Counting cross-overs: characterizing meiotic recombination in mammals

Until recently, most of our understanding of meiotic recombination has come from studies of lower eukaryotes. However, over the past few years several components of the mammalian meiotic recombination pathway have been identified, and new molecular and cytological approaches to the analysis of mammalian meiosis have been developed. In this review, we discuss recent advances in three areas: the application of new techniques to study genome-wide levels of recombination in individual meioses; studies analyzing temporal aspects of the mammalian recombination pathway; and studies linking the genesis of human trisomies to alterations in meiotic exchange patterns.     

Application of molecular techniques to the study of human physiological variation

Physiological differences between human populatios, manifest as apparent adaptation to extreme environments, as differences in patterna of growth and development, and as differences in prevalence of common disease, are a mofer subject area of human biology. The approach of the geneticist interested in understanding these difference has been largely that of the proverbial drunk looking for has lost keys. We have characterized those genes whose products are easily assayed from peripheral blood and sought post hoc statistical evidence for a relationship between gene frequencies and physiological variables. With few exceptions, this approach has failed to provide convincing evidence of the role of genes in determining physiological differences between populations. This is not unexpected since the genes which our knowledge of physiology would predict to be important in determining physiological homeostasis. Recent advances in molecular biology (particularly in the ability to isolate and clone specific genes and to use these as probes to detect genomic variation—polymorphism—associated with these genes in human populations) and advances in our ability to resolve and visualize genetic variation in protein products provide the tools for a direct evaluation of the role of specific gene in determining physiological differences between populations. I will review the “candidate genes” available for the study of several physiological systems and present preliminary data from my laboratory related to genetic determinants of lipid metabolism.     

Human cell models for genetic engineering

Technological advances made in molecular biology and culture of human and other mammalian cells in vitro have increased the ability to introduce functional genes into a variety of cell types. Subgenomic components of both DNA and RNA viruses are used as the eukaryotic components of many recombinant DNA vector constructs which are used for transfection either by a variety of methods to facilitate DNA uptake or as virions. This paper reviews some of the factors involved in expression, persistence, and recombination of the introduced genes. Also considered are the use of DNA transfection models to study human cancer or other diseases, the current status of gene therapy, and the use of human cells to produce biologicals. The need to better develop well-characterized human cell culture models from multiple organ sites and various cell types is a recurrent theme for all of these applications, including the role that such cells will play in understanding the functions for genes which will be identified in the human genome cloning project. With continued study and a better understanding of pathophysiology at the cellular and molecular levels should come the logical next step of adding genetically engineered human cells to the repertoire of acceptable biomedical technologies.     

Towards subtlety: understanding the role of Toll-like receptor signaling in susceptibility to human infections

Recent years have seen a dramatic improvement in our understanding of the role of innate immunity, and particularly Toll-like receptor (TLR) signaling, in human host defense. Appreciation of how defects in human TLR signaling enhance susceptibility to infection began with the identification of patients with monogenic immunodeficiencies, such as hypohydrotic ectodermal dysplasia with immunodeficiency and IRAK4 deficiency. Empowered by technological advances in genotyping and bioinformatics, we are now beginning to appreciate how common genetic variation in the genes controlling the innate immune response alters infectious susceptibility in a subtle but specific fashion. This review highlights the mechanisms of infectious susceptibility that result from complex interactions between the genetically variable host and microbe and explores how this new knowledge may ultimately translate into better care for our patients.     

Endocannabinoids and cannabinoid receptor genetics

This review presents the remarkable advances that have been achieved in marijuana (cannabinoid) research, with the discovery of specific receptors and the existence of naturally occurring cannabis-like substances in the human body and brain. The last decade has seen more rapid progress in marijuana research than any time in the thousands of years that marijuana has been used by humans, particularly in cannabinoid genomics. The cDNA and genomic sequences encoding G protein-coupled cannabinoid receptors (Cnrs) from several species have now been cloned. Endogenous cannabinoids (endocannabinoids), synthetic and hydrolyzing enzymes and transporters that define neurochemically-specific cannabinoid brain pathways have been identified. Endocannabinoid lipid signaling molecules alter activity at G protein-coupled receptors (GPCR) and possibly at anandamide-gated ion channels, such as vanilloid receptors. Availability of increasingly-specific CB1 and CB2 Cnr antagonists and of CB1 and CB2 Cnr knockout mice have increased our understanding of these cannabinoid systems and provides tantalizing evidence for even more G protein-coupled Cnrs. Initial studies of the Cnr gene structure, regulation and polymorphisms whet our appetite for more information about these interesting genes, their variants and roles in vulnerabilities to addictions and other neuropsychiatric disorders. Behavioral studies of cannabinoids document the complex interactions between rewarding and aversive effects of these drugs. Pursuing cannabinoid-related molecular, pharmacological and behavioral leads will add greatly to our understanding of endogenous brain neuromodulator systems, abused substances and potential therapeutics. This review of CB1 and CB2 Cnr genes in human and animal brain and their neurobiological effects provide a basis for many of these studies. Therefore, understanding the physiological cannabinoid control system in the human body and brain will contribute to elucidating this natural regulatory mechanism in health and disease.     

PRR-signaling pathways: Learning from microbial tactics

Recognition of bacterial pathogens by the mammalian host relies on the induction of early innate immune responses initiated by the activation of pattern-recognition receptors (PRRs) upon sensing of their cognate microbe-associated-patterns (MAMPs). Successful pathogens have evolved to intercept PRR activation and signaling at multiple steps. The molecular dissection of the underlying mechanisms revealed many of the basic mechanisms used by the immune system.Here we provide an overview of the different strategies used by bacterial pathogens and commensals to subvert and reprogram PPR-mediated innate immune responses. A particular attention is given to recent discoveries highlighting novel molecular details of the host inflammatory response in mammalian cells and current advances in our understanding of the interaction of commensals with PRR-mediated responses.     

Metal ion transporters in mammals: structure, function and pathological implications

Despite the importance of metal ions in several catalytic functions, there has been, until recently, little molecular information available on the mechanisms whereby metal ions are actively taken up by mammalian cells. The classical concept for iron uptake into mammalian cells has been the endocytosis of transferrin-bound Fe³⁺ by the transferrin receptor. Studies with hypotransferrinaemic mice revealed that in the intestine mucosal transferrin is derived from the plasma and that its presence is not required in the intestinal lumen for dietary iron absorption. This suggests that, at least in the intestine, other non-receptor-mediated uptake systems exist. The molecular identification of metal ion transporters is of great importance, in particular since an increasing number of human diseases are thought to be related to disturbances in metal ion homeostasis, including metal ion overload and deficiency disorders (i.e. anaemia, haemochromatosis, Menkes disease, Wilson's disease), and neurodegenerative diseases (i.e. Alzheimer's, Friedreich's ataxia and Parkinson's diseases). Furthermore, susceptibilities to mycobacterial infections are caused by metal ion transporter defects. The pathological implications of disturbed metal ion homeostasis confirm the vital roles these metal ions play in the catalytic function of many enzymes, in gene regulation (zinc-finger proteins), and in free radical homeostasis. Recent insights have significantly advanced our knowledge of how metal ions are taken up or released by mammalian cells. The purpose of this review is to summarize these advances and to give an overview on the growing number of mammalian metal ion transporters.     

Ontogeny and expansion of human natural killer cells: clinical implications

Our knowledge of NK cells and their critical role in the innate immune system has increased enormously since their discovery several decades ago. However, it is only within the last 10 years that rational cytokine therapies, such as those utilizing low doses of IL-2, have been successful in expanding NK cells in patients with cancer and/or immunodeficiency. Such experiences in vivo have highlighted the importance of basing immunotherapeutic strategies on the known cellular and molecular properties of the targeted cell population. Recent advances in our understanding of the physiologic factors and events that orchestrate NK cell ontogeny, including IL-15 and receptor tyrosine kinase ligands to c-kit and flt3, provide novel therapeutic possibilities for cytokine therapy. This review summarizes our current understanding of human NK cell ontogeny, and links this knowledge to ongoing and future clinical strategies for the endogenous expansion of NK cells in patients with cancer and/or immunodeficiency.     

Ontogeny and Expansion of Human Natural Killer Cells: Clinical Implications

Our knowledge of NK cells and their critical role in the innate immune system has increased enormously since their discovery several decades ago. However, it is only within the last 10 years that rational cytokine therapies, such as those utilizing low doses of IL-2, have been successful in expanding NK cells in patients with cancer and/or immunodeficiency. Such experiences in vivo have highlighted the importance of basing immunotherapeutic strategies on the known cellular and molecular properties of the targeted cell population. Recent advances in our understanding of the physiologic factors and events that orchestrate NK cell ontogeny, including IL-15 and receptor tyrosine kinase ligands to c-kit and fit3, provide novel therapeutic possibilities for cytokine therapy. This review summarizes our current understanding of human NK cell ontogeny, and links this knowledge to ongoing and future clinical strategies for the endogenous expansion of NK cells in patients with cancer and/or immunodeficiency.     

Current and novel therapeutics in the treatment of systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a complex autoimmune disease with significant clinical heterogeneity. Recent advances in our understanding of the genetic, molecular, and cellular bases of autoimmune diseases and especially SLE have led to the application of novel and targeted treatments. Although many treatment modalities are effective in lupus-prone mice, the situation is more complex in human subjects. This article reviews the general approach to the therapy of SLE, focusing on current approved therapies and novel approaches that might be used in the future.     

Muscle unloading: A comparison between spaceflight and ground‐based models

Prolonged unloading of skeletal muscle, a common outcome of events such as spaceflight, bed rest and hindlimb unloading, can result in extensive metabolic, structural and functional changes in muscle fibres. With advancement in investigations of cellular and molecular mechanisms, understanding of disuse muscle atrophy has significantly increased. However, substantial gaps exist in our understanding of the processes dictating muscle plasticity during unloading, which prevent us from developing effective interventions to combat muscle loss. This review aims to update the status of knowledge and underlying mechanisms leading to cellular and molecular changes in skeletal muscle during unloading. We have also discussed advances in the understanding of contractile dysfunction during spaceflights and in ground‐based models of muscle unloading. Additionally, we have elaborated on potential therapeutic interventions that show promising results in boosting muscle mass and strength during mechanical unloading. Finally, we have identified key gaps in our knowledge as well as possible research direction for the future.     

Islet–immune interactions in type 1 diabetes: the nexus of beta cell destruction

Recent studies in Type 1 Diabetes (T1D) support an emerging model of disease pathogenesis that involves intrinsic β-cell fragility combined with defects in both innate and adaptive immune cell regulation. This combination of defects induces systematic changes leading to organ-level atrophy and dysfunction of both the endocrine and exocrine portions of the pancreas, ultimately culminating in insulin deficiency and β-cell destruction. In this review, we discuss the animal model data and human tissue studies that have informed our current understanding of the cross-talk that occurs between β-cells, the resident stroma, and immune cells that potentiate T1D. Specifically, we will review the cellular and molecular signatures emerging from studies on tissues derived from organ procurement programs, focusing on in situ defects occurring within the T1D islet microenvironment, many of which are not yet detectable by standard peripheral blood biomarkers. In addition to improved access to organ donor tissues, various methodological advances, including immune receptor repertoire sequencing and single-cell molecular profiling, are poised to improve our understanding of antigen-specific autoimmunity during disease development. Collectively, the knowledge gains from these studies at the islet–immune interface are enhancing our understanding of T1D heterogeneity, likely to be an essential component for instructing future efforts to develop targeted interventions to restore immune tolerance and preserve β-cell mass and function.     

Marine invertebrate eco-genotoxicology: a methodological overview

The last 25 years have seen major advances in the field of mammalian genotoxicology, particularly with the advent of molecular methods, some of which have spilled over into the relatively new field of eco-genotoxicology, which aims to evaluate the impact of contaminants on the natural biota. Unlike mammalian genotoxicology, where the focus is centred on a limited number of model species, efforts in the marine field have generally lacked coordination and focus, with the result that progress has been somewhat slow and fragmented. However, it is recognized that at the DNA and chromosome levels, marine invertebrates express qualitatively similar types of induced damage to that found in higher organisms (e.g. point mutations, strand breaks and chromosomal aberrations). Given that many of these species (bivalve molluscs, crustaceans, polychaete worms, etc.) are linked directly or indirectly to the human food chain, this is an important reason why one should be concerned about their exposure to environmental mutagens and carcinogens, particularly as many of these organisms have the capacity to (i) transform these agents to biologically active metabolites and (ii) accumulate toxicants in their cells and tissues at concentrations several orders of magnitude above that found in the environment. This review covers the advantages and limitations of those cytogenetic and molecular assays that have been used to address the question of genotoxicity in the cells and early life stages of selected marine invertebrate species. It concludes with the recommendation for the adoption of standardized test procedures, leading to a tiered approach in future eco-genotoxicity testing.     

Viral interactions with the host and microbiota in the intestine

This review explores the recent advances that have been made in our understanding of host viral interactions in the intestine. Technical advances have allowed the initial definition of intestinal viromes in a number of species including humans. Important advances in our knowledge of the host response to viral infection have shown that interferon lambda has a role that is unique from type I interferons in the intestine. Lastly, our understanding of virally induced phenotypes has expanded through new studies that show bacteria can play an important role in the outcome of viral infection in the intestine.     

Intimate relationships with their neighbors: tales of stem cells in Drosophila reproductive systems.

Stem cells have the unique potential to self-renew and to supply differentiated cells that replenish lost cells throughout an organism's lifetime. This unique property makes stem cells powerful therapeutic tools for future regenerative medicine. However, the molecular mechanisms of stem cell regulation are still poorly understood in many stem cell systems. Stem cell function has been shown recently to be controlled by concerted actions of extrinsic signals from its regulatory niche and intrinsic factors inside the stem cell. Stem cells in the Drosophila reproductive systems provide excellent models to understand the fundamental mechanisms underlying stem cell regulation, including the relationships between stem cells and their niches. Within the past few years, much progress in understanding stem cells in Drosophila has been made, and the knowledge gained from studying these stem cells greatly advances our understanding of stem cells in other systems, including humans. In this review, we summarize the recent progress and describe future challenges in understanding the molecular mechanisms controlling stem cell self-renewal, division, and differentiation in the Drosophila reproductive systems.