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With the advancement of single-cell separation techniques and high-throughput sequencing platforms, single-cell RNA-sequencing (scRNA-seq) has emerged as a vital technology for understanding tissue and organ systems at cellular resolution. Through transcriptional analysis, it is possible to characterize unique or rare cell types, interpret their interactions, and reveal novel functional states or shifts in developmental stages. As such, this technology is uniquely suited for studying the cells within the human ovary. The ovary is a cellularly heterogeneous organ that houses follicles, the reproductive and endocrine unit that consists of an oocyte surrounded by hormone-producing support cells, as well as many other cell populations constituting stroma, vasculature, lymphatic, and immune components. Here we review studies that have utilized scRNA-seq technology to analyze cells from healthy human ovaries and discuss the single-cell isolation techniques used. We identified two overarching applications for scRNA-seq in the human ovary. The first applies this technology to investigate transcriptional differences in oocytes/eggs from patients undergoing in vitro fertilization treatments to ultimately improve clinical outcomes. The second utilizes scRNA-seq for the pursuit of creating a comprehensive single-cell atlas of the human ovary. The knowledge gained from these studies underscores the importance of scRNA-seq technologies in unlocking a new biological understanding of the human ovary.

The Gap Park Self-Harm Minimisation Masterplan project is a collaborative attempt to address jumping suicides at Sydney’s Gap Park through means restriction, encouraging help-seeking, and increasing the likelihood of third-party intervention. We used various data sources to describe the Masterplan project’s processes, impacts, and outcomes. There have been reductions in reported jumps and confirmed suicides, although the trends are not statistically significant. There has been a significant increase in police call-outs to intervene with suicidal people who have not yet reached the cliff’s edge. The collaborative nature of the Masterplan project and its multifaceted approach appear to be reaping benefits.

The modulation of the startle response (SR) by threatening stimuli (fear-potentiated startle; FPS), is a proposed endophenotype for disorders of the fearful-fearlessness spectrum. FPS has failed to show evidence of heritability, raising concerns. However, the metrics used to index FPS – and, importantly, other conditional phenotypes that are dependent on a baseline – may not be suitable for the approaches used in genetic epidemiology studies. Here we evaluated multiple metrics of FPS in a population-based sample of pre-adolescent twins (N = 569 from 320 twin pairs, M(age) = 11.4) who completed a fear-conditioning paradigm with airpuff-elicited SR on two occasions (~1 month apart). We applied univariate and multivariate biometric modeling to estimate the heritability of FPS using several proposed standardization procedures. This was extended with data simulations to evaluate biases in heritability estimates of FPS (and similar metrics) under various scenarios. Consistent with previous studies, results indicated moderate test-retest reliability (r = .59) and heritability of the overall SR (h(2) = 34%) but poor reliability and virtually no unique genetic influences on FPS when considering a raw or standardized differential score that removes baseline SR. Simulations demonstrated that the use of differential scores introduces bias in heritability estimates relative to jointly analyzing baseline SR and FPS in a multivariate model. However, strong dependency of FPS on baseline levels make unique genetic influences virtually impossible to detect regardless of methodology. These findings indicate that FPS and other conditional phenotypes may not be well-suited to serve as endophenotypes unless such co-dependency can be disentangled.

The traditional method of reporting health care costs in the United States is by setting or payer, such as the National Health Expenditure Accounts (NHEA), produced by the Centers for Medicare & Medicaid Services (CMS) (Centers for Medicare and Medicaid Services 2009). International comparisons have predominantly focused on overall trends, financing agent, provider, and function, and often provide limited information on specific consumer encounters and spending (Anderson and Hussey 2001; Orosz and Morgan 2004;). The Organization for Economic Co-Operation and Development has examined person-centered categories of health spending. These methods are useful for trends overall and by setting and payer but do not predominantly focus on how consumers actually interact with the health care system. We aimed to combine traditional methods of accounting with a patient-centered viewpoint to more fully identify the underlying factors driving costs and potential levers for efficient allocation of health care resources. The Medical Expenditure Panel Survey (MEPS) collects data on conditions, services, individual characteristics, setting, and payers (Agency for Healthcare Research and Quality 2009). Our objective was to classify all U.S. health care expenditures into patient-centered categories that capture health events in a manner that is closer than conventional methods (e.g., by setting or payer) to how consumers experience and consider their health and health care spending. We used MEPS data to classify medical expenses in 2007 into seven “patient-centered care” categories. This method of examining expenditures can complement traditional methods to provide additional insight into spending. This categorization may better identify the drivers of health care expenditures and suggest more focused policy solutions to improve the value and efficiency of the health care system.

Immunogold staining in conjunction with TEM was used to observe C3 adsorption from plasma in relation to the underlying titanium structure of thermal, anodic, and electropolished oxides. Heat treatments and oxide thickness were found to have no significant effect on the adsorption behavior of C3, while surface oxide type possibly has. Surface concentration of C3 was found to be time- and plasma concentration-dependent. Evidence is given for the possible involvement of C3 in protein exchange, i.e., the Vroman effect. Diluted plasma resulted in a random distribution of gold colloids, whereas clustering occurred with undiluted plasma. Although C3 concentrations present on grain boundaries followed the same trend as that found on the surface, C3 was found to have a higher grain boundary than bulk concentration for 0.1% plasma.

Following administration of high levels of vitamin E, plasma tocopherol levels in mature humans and monkeys continued to increase with time. In man plasma tocopherol levels were higher when tocopherol rather than tocopheryl acetate was given. Mature female rats that had been maintained on laboratory chow were put on the diet supplemented with either 1,000 or 10,000 mg/kg of all-rac-alpha-tocopheryl acetate. All tissues analyzed (plasma, platelets, liver, red blood cells, adipose tissue, heart, lung, skeletal muscle, and brain) continued to increase in tocopherol content for the duration of the supplement (20 weeks). It was concluded that it is difficult to saturate tissue with tocopherol and that not only the level, but the duration, of supplementation with vitamin E influences the concentration of vitamin E in all tissues. Liver and adipose tissue both accumulated tocopherol at a very rapid rate compared to other tissues, but once the chow diet was resumed, liver tocopherol decreased very rapidly and adipose tissue decreased very slowly. The studies suggest that at least for a short time period, the liver is the major available storage organ for tocopherol. When animals were put back on the unsupplemented chow diet, platelet and plasma levels returned to baseline values within 1 week, and red blood cells, heart, muscle, lung, and brain within 4 weeks.

Highly reactive molecules called free radicals can cause tissue damage by reacting with polyunsaturated fatty acids in cellular membranes, nucleotides in DNA, and critical sulfhydryl bonds in proteins. Free radicals can originate endogenously from normal metabolic reactions or exogenously as components of tobacco smoke and air pollutants and indirectly through the metabolism of certain solvents, drugs, and pesticides as well as through exposure to radiation. There is some evidence that free radical damage contributes to the etiology of many chronic health problems such as emphysema, cardiovascular and inflammatory diseases, cataracts, and cancer. Defenses against free radical damage include tocopherol (vitamin E), ascorbic acid (vitamin C), beta-carotene, glutathione, uric acid, bilirubin, and several metalloenzymes including glutathione peroxidase (selenium), catalase (iron), and superoxide dismutase (copper, zinc, manganese) and proteins such as ceruloplasmin (copper). The extent of tissue damage is the result of the balance between the free radicals generated and the antioxidant protective defense system. Several dietary micronutrients contribute greatly to the protective system. Based on the growing interest in free radical biology and the lack of effective therapies for many of the chronic diseases, the usefulness of essential, safe nutrients in protecting against the adverse effects of oxidative injury warrants further study.

A new type of image, existing only at field strengths below the denaturation field strengths of molecules, has been discorvered. This type of image has structure, is not symmetrical and thus differs from previously reported low field strength images. The possibility that macromolecules adsorbed on tip surfaces produce such structured images has been exhaustively investigated. The result is that no observation has been found which disproves this hypothesis and many tests conducted in such attempts yielded correlations consistent with this hypothesis. It is therefore concluded that it is highly probable that biomolecules produce structured images and that it is highly improbable that these correlations represent a chance event. On the other hand, these correlations may be due to some cause unknown to the authors; but we consider this possibility to be unlikely. Some micrographs have been obtained which provide a reasonable basis for the hope that tertiary structure information may be defrived from low field strength imaging of partially embedded biomolecules or of biomolecules that are resistant to field denaturation during imaging. Information may presently be obtained from analysis of low field strength ion micrographs about the size and shape of some biomolecules.

An adaptation of the needle biopsy procedure of Beynen and Katan for human adipose tissue, which yields 2-10 mg adipose samples, is described and evaluated. Micromethods are presented for the analysis of alpha-tocopherol, cholesterol and fatty acids in each adipose specimen. The needle biopsy procedure, which uses a Vacutainer to create suction, is compared with a punch biopsy method. The needle biopsy is rapid (6 samples/hr), simple and unobjectionable to the subjects, and provides samples with reproducible ratios of cholesterol and alpha-tocopherol. Unlike the punch biopsy, the needle biopsy reliably obtains specimens with a lipid composition typical of adipocytes. The needle biopsy method is adaptable to nutritional studies of tocopherol and fatty acid metabolism in adipose, and to studies of hazardous compounds stored in adipose. The linoleic acid content of adipose from residents of the West Coast was found to be considerably higher than values reported earlier. The adipose fatty acid data indicate an increase in human adipose linoleate when compared with earlier reports and suggest a trend toward increasing linoleic acid in the American diet.

A cleavage technique was used to measure the surface free energy and the grain boundary energy for twist boundaries of varying twist angles in potassium chloride. The results for the surface energy and its temperature dependence are in good agreement with theory and experiment. Based in part on this agreement and the measured misfit orientation dependence, the grain boundary energy values obtained are concluded to be reasonable.

The addition of polyunsaturated fatty acids (PUFA) to diets deficient in vitamin E and other effective antioxidants results in a variety of symptoms in animals. For example, the feeding of such diets to rats results in muscular dystrophy, testis degeneration, dental depigmentation, brown discoloration of the fat and and uterus and creatinuria Similar diets fed to rabbits and ruminants results in muscular dystrophy. In chickens the symptoms observed are encephalomalacia, lowered egg production, and poor hatchability. The addition of PUFA to diets is known to result in the destruction of vitamin E in the diet or in the tissues of animals as a result of free radicals produced during the autooxidation of the PUFA. However, in several studies, this possible explanation for the development of vitamin E deficiency symptoms has been made untenable. In such studies the more likely explanation for development of symptoms is thein vivo peroxidation of PUFA in the tissues of animals following incorporation of large amounts of PUFA in lipid structures and depletion from the tissues of vitamin E and other biologically effective antioxidants.

Encephalomalacia was induced in chickens more than 64 days old by feeding a high linoleic acid diet with an antioxidant (ethoxyquin) for 64 days and then deleting the antioxidant.