Sensitive and tolerant raters differ in their magnitude estimates of extremely violent behaviors.
A 2007 study indicated severity of violence of several behaviors can be rated consistently, such that an individual's average rating across the behaviors can serve as a measure of sensitivity to violence. However, violence-sensitive and violence-tolerant raters in that study gave similar high scale ratings of severity of violence for extremely violent behaviors such as murder and stabbing. In the present study an open-ended magnitude estimation response scale was used by 27 participants to examine impressions of the severity of violence of 38 behaviors. There was a positive correlation between estimates of severity for extreme physical violence and estimates for lower severity of violence. This result indicates that the earlier finding of apparent equivalence of sensitive and tolerant individuals did not signify agreement about extremely violent behaviors but was a ceiling effect arising from the closed-ended response scale used. Violence-sensitive and violence-tolerant individuals seem to have different attitudes toward all types of violence.


Zinc determines dynamical properties and aggregation kinetics of human insulin.
Protein aggregation is a widespread process leading to deleterious consequences in the organism, with amyloid aggregates being important not only in biology but also for drug design and biomaterial production. Insulin is a protein largely used in diabetes treatment, and its amyloid aggregation is at the basis of the so-called insulin-derived amyloidosis. Here, we uncover the major role of zinc in both insulin dynamics and aggregation kinetics at low pH, in which the formation of different amyloid superstructures (fibrils and spherulites) can be thermally induced. Amyloid aggregation is accompanied by zinc release and the suppression of water-sustained insulin dynamics, as shown by particle-induced x-ray emission and x-ray absorption spectroscopy and by neutron spectroscopy, respectively. Our study shows that zinc binding stabilizes the native form of insulin by facilitating hydration of this hydrophobic protein and suggests that introducing new binding sites for zinc can improve insulin stability and tune its aggregation propensity.


A Spiking Neural Simulator Integrating Event-Driven and Time-Driven Computation Schemes Using Parallel CPU-GPU Co-Processing: A Case Study.
Time-driven simulation methods in traditional CPU architectures perform well and precisely when simulating small-scale spiking neural networks. Nevertheless, they still have drawbacks when simulating large-scale systems. Conversely, event-driven simulation methods in CPUs and time-driven simulation methods in graphic processing units (GPUs) can outperform CPU time-driven methods under certain conditions. With this performance improvement in mind, we have developed an event-and-time-driven spiking neural network simulator suitable for a hybrid CPU-GPU platform. Our neural simulator is able to efficiently simulate bio-inspired spiking neural networks consisting of different neural models, which can be distributed heterogeneously in both small layers and large layers or subsystems. For the sake of efficiency, the low-activity parts of the neural network can be simulated in CPU using event-driven methods while the high-activity subsystems can be simulated in either CPU (a few neurons) or GPU (thousands or millions of neurons) using time-driven methods. In this brief, we have undertaken a comparative study of these different simulation methods. For benchmarking the different simulation methods and platforms, we have used a cerebellar-inspired neural-network model consisting of a very dense granular layer and a Purkinje layer with a smaller number of cells (according to biological ratios). Thus, this cerebellar-like network includes a dense diverging neural layer (increasing the dimensionality of its internal representation and sparse coding) and a converging neural layer (integration) similar to many other biologically inspired and also artificial neural networks.


Field evaluation of an ELISA to detect antibody in leprosy patients and their contacts.
Previous studies have detected circulating antibody in leprosy using a variety of difficult laboratory methods. We have developed a simpler method for detecting antibody by ELISA, using autoclaved Mycobacterium smegmatis as the antigen. Evaluation was performed on eluates from 25 microliter aliquots of finger-prick blood dried on filter-paper disks in two high-incidence populations in Ponape, Micronesia. Among 228 nonleprosy cases bled in 1980 and rebled and re-examined in 1982: a) for those who had been ELISA positive two years earlier, the leprosy attack rate during the intervening two years was at least twice as high as among those who had been negative, and we estimate that shortening the screening interval to one year plus doing confirmatory retests on new sero-converters would increase the relative risk (or "predictive power") to over sixfold, including all impending multibacillary cases; b) elevated antibody levels were detected up to two years prior to clinical onset of disease in 70% of new cases; and c) both asymptomatic conversion (rising titer) and reversion (falling titer) were observed. Among 150 biopsy-proven cases, ELISA results suggest that fall of titer in most uncomplicated paucibacillary cases was rapid (months), but in multibacillary cases was more gradual (years), probably paralleling responses to treatment with titers rising in reactivation. These results suggest that this technique, with an improved antigen, may be useful in leprosy control programs, both for detecting candidates for preventive treatment and for following responses to therapy.


Habitat fragmentation causes bottlenecks and inbreeding in the European tree frog (Hyla arborea).
A genetic study of the European tree frog, Hyla arborea, in Denmark was undertaken to examine the population structure on mainland Jutland and the island of Lolland after a period of reduction in suitable habitat and population sizes. The two regions have experienced the same rate of habitat loss but fragmentation has been more severe on Lolland. Genetic variation based on 12 polymorphic DNA microsatellites was analysed in 494 tree frogs sampled from two ponds in Jutland and 10 ponds on Lolland. A significant overall deviation from Hardy-Weinberg expectations could be attributed to three ponds, all on Lolland. This was most probably caused by an inbreeding effect reducing fitness, which was supported by the observed significant negative correlation between larva survival and mean F(IS) value and mean individual inbreeding coefficient. A significant reduction in genetic variation (bottleneck) was detected in most of the ponds on Lolland. Population-structure analysis suggested the existence of at least 11 genetically different populations, corresponding to most of the sampled population units. The results indicated that the populations were unique genetic units and could be used to illustrate the migration pattern between newly established ponds arisen either by natural colonization of tree frogs or by artificial introduction. A high degree of pond fidelity in the tree frogs was suggested. A severe fragmentation process reducing population size and fitness within some of the populations probably caused the significant reduction in genetic variation of tree frog populations on Lolland.


A medieval fallacy: the crystalline lens in the center of the eye.
To determine whether, as most modern historians have written, ancient Greco-Roman authors believed the crystalline lens is positioned in the center of the eye. Historians have written that statements about cataract couching by Celsus, or perhaps Galen of Pergamon, suggested a centrally located lens. Celsus specifically wrote that a couching needle placed intermediate between the corneal limbus and the lateral canthus enters an empty space, presumed to represent the posterior chamber. Ancient ophthalmic literature was analyzed to understand where these authors believed the crystalline lens was positioned. In order to estimate where Celsus proposed entering the eye during couching, we prospectively measured the distance from the temporal corneal limbus to the lateral canthus in 30 healthy adults. Rufus of Ephesus and Galen wrote that the lens is anterior enough to contact the iris. Galen wrote that the lens equator joins other ocular structures at the corneoscleral junction. In 30 subjects, half the distance from the temporal corneal limbus to the lateral canthus was a mean of 4.5 mm (range: 3.3-5.3 mm). Descriptions of couching by Celsus and others are consistent with pars plana entry of the couching needle. Anterior angulation of the needle would permit contact of the needle with the lens. Ancient descriptions of anatomy and couching do not establish the microanatomic relationships of the ciliary region with any modern degree of accuracy. Nonetheless, ancient authors, such as Galen and Rufus, clearly understood that the lens is located anteriorly. There is little reason to believe that Celsus or other ancient authors held a variant understanding of the anatomy of a healthy eye. The notion of the central location of the lens seems to have arisen with Arabic authors in 9th century Mesopotamia, and lasted for over 7 centuries.


A Questionnaire for Rating Health-related Quality of Life.
Translations of instruments for measuring quality of life developed in certain, mostly more developed, parts of the world usually do not cover regionally specific aspects of health-related quality of life, even after transcultural validation. The aim of this study was to develop and validate a reliable questionnaire in Serbian, Croatian, Bosnian, and Montenegrin languages suitable for measuring health-related quality of life in adults. The study was of a cross-sectional type, assessing the reliability and validity of a newly developed questionnaire for measuring health-related quality of life (HRQoL) in adults residing in western Balkan states (WB-HRQoL). It was conducted on a sample of 489 adults from Serbia, Croatia, Bosnia & Herzegovina, and Montenegro, with a mean age of 52.2±14.4 years and a male/female ratio of 195/294 (39.9%/60.1%). The definitive version of the WB-HRQoL scale with 19 items showed very good reliability, with Cronbach's alpha 0.905. The scale was temporally stable, and satisfactory results were obtained for divergent and convergent validity tests. Exploratory factorial analysis brought to the surface four domains of health-related quality of life, namely the physical, psychical, social, and environmental. The WB-HRQoL scale is a reliable and valid generic instrument for measuring HRQoL that takes into account the cultural specifics of the western Balkan region.


The management of adult growth hormone deficiency syndrome.
In patients with hypopituitarism, growth hormone deficiency (GHD) is almost inevitably present. Lack of other pituitary hormones may require prompt replacement, but lack of growth hormone (GH) is also associated with several abnormalities. Growth hormone treatment safely improves or normalizes many of these deviations. The aberrations include low bone mass and increased risk of fractures, abnormal body composition in terms of increased fat mass, and reduced lean body mass resulting in reduced muscle mass and strength. The decreased exercise capacity in GHD may be influenced by impaired cardiac performance and heat intolerance. Increased abdominal fat results in metabolic disturbancies such as reduced insulin sensitivity and hyperlipidemia, leading to an increased risk of cardiovascular diseases. Patients with hypopituitarism who have had relevant hormones replaced except GH have increased mortality due to, for example, cardiovascular diseases. Thus, it is important to diagnose GHD, which requires precise diagnostic criteria and methods. Dynamic testing of GH secretion with insulin tolerance test or arginine + GH releasing hormone can be used. Patients with childhood onset GHD should be re-evaluated if GH therapy is maintained into adulthood. Although GH replacement therapy improves the patophysiological manifestations of GHD and quality of life, it is unclear whether mortality is reduced.


Genomic Exploration of the Brain in People Infected with HIV-Recent Progress and the Road Ahead.
The adult human brain harbors billions of microglia and other myeloid and lymphoid cells highly susceptible to HIV infection and retroviral insertion into the nuclear DNA. HIV infection of the brain is important because the brain is a potentially large reservoir site that may be a barrier to HIV cure strategies and because infection can lead to the development of HIV-associated neurocognitive disorder. To better understand both the central nervous system (CNS) reservoir and how it can cause neurologic dysfunction, novel genomic, epigenomic, transcriptomic, and proteomic approaches need to be employed. Several characteristics of the reservoir are important to learn, including where the virus integrates, whether integrated proviruses are intact or defective, whether integrated proviruses can be reactivated from a latent state to seed ongoing infection, and how this all impacts brain function. Here, we discuss similarities and differences of viral integration sites between brain and blood and discuss evidence for and against the hypothesis that in the absence of susceptible T-lymphocytes in the periphery, the virus housing in the infected brain is not able to sustain a systemic infection. Moreover, microglia from HIV + brains across a wide range of disease severity appear to share one type of common alteration, which is defined by downregulated expression, and repressive chromosomal compartmentalization, for microglial genes regulating synaptic connectivity. Therefore, viral infection of the brain, including in immunocompetent cases with near-normal levels of CD4 blood lymphocytes, could be associated with an early disruption in microglia-dependent neuronal support functions, contributing to cognitive and neurological deficits in people living with HIV.


Development and validation of the Delaying Gratification Inventory.
Deficits in gratification delay are associated with a broad range of public health problems, such as obesity, risky sexual behavior, and substance abuse. However, 6 decades of research on the construct has progressed less quickly than might be hoped, largely because of measurement issues. Although past research has implicated 5 domains of delay behavior, involving food, physical pleasures, social interactions, money, and achievement, no published measure to date has tapped all 5 components of the content domain. Existing measures have been criticized for limitations related to efficiency, reliability, and construct validity. Using an innovative Internet-mediated approach to survey construction, we developed the 35-item 5-factor Delaying Gratification Inventory (DGI). Evidence from 4 studies and a large, diverse sample of respondents (N = 10,741) provided support for the psychometric properties of the measure. Specifically, scores on the DGI demonstrated strong internal consistency and test-retest reliability for the 35-item composite, each of the 5 domains, and a 10-item short form. The 5-factor structure fit the data well and had good measurement invariance across subgroups. Construct validity was supported by correlations with scores on closely related self-control measures, behavioral ratings, Big Five personality trait measures, and measures of adjustment and psychopathology, including those on the Minnesota Multiphasic Personality Inventory-2-Restructured Form. DGI scores also showed incremental validity in accounting for well-being and health-related variables. The present investigation holds implications for improving public health, accelerating future research on gratification delay, and facilitating survey construction research more generally by demonstrating the suitability of an Internet-mediated strategy.


Molecular cloning and sequencing of salmon gonadotropin beta subunit.
Gonadotropin (GTH) was purified from the pituitaries of the Pacific chinook salmon using a combination of stepwise ethanol precipitation and concanavalin-A affinity chromatography. The alpha and beta subunits were dissociated and fractionated by C-18 reverse-phase high-performance liquid chromatography with a 0.1% trifluoroacetic acid/acetonitrile gradient. An enriched cDNA library was screened for the beta-GTH gene(s) using two synthetic oligonucleotides based on partial protein data. A positive, full-size clone (E3) was identified and sequenced. It contains 657 base pairs and codes for a 142-amino-acid precursor protein. The mature salmon beta-GTH (119 amino acids) is structurally homologous to human luteinizing hormone and chorionic gonadotropin. The effect of testosterone implantation on pituitary GTH and beta-GTH mRNA was examined with radioimmunoassay and Northern blot analysis. There was a corresponding increase in both the pituitary GTH and mRNA levels.


Assessment of Oxidative Stress Genes SOD2 and SOD3 Polymorphisms Role in Human Colorectal Cancer.
The aim of this study was to ascertain the oxidative stress genes SOD2 and SOD3 polymorphisms in patients with colorectal cancer and to assess the possible involvement of these polymorphisms that might increase the risk for patients to develop malignant intestinal tumors. A total number of 306 subjects were divided into two groups (109 colorectal cancer patients as the study group and 197 normal healthy individuals as the control group).We genotyped two polymorphisms, SOD2 - 201A>G (rs4880) and SOD3 - 896C>G (rs1799895), by allelic discrimination, with TaqMan RT-PCR specific probes. No significant differences were found with either of the polymorphisms when comparing the association between them and an increased risk of developing colorectal tumors. In Romanian population, the risk of developing colorectal cancer is not increased by SOD2 and SOD3 polymorphisms.


Psychological Distress of Ethnically Diverse Adult Caregivers in the California Health Interview Survey.
This study examined factors associated with psychological distress for culturally diverse family caregivers using a population-based sample. Data were analyzed from the 6634 caregivers of adults (i.e. elderly as well as non-elderly) who self-reported as non-Hispanic White, Mexican, Chinese, or Vietnamese in the 2009 California Health Interview Survey. Simple and multiple regression analyses were conducted to assess the potential influence of race/ethnicity, caregiving context, and social contextual variables on psychological distress. Analyses that included moderators showed that while more education was associated with less distress for White caregivers, it was associated with more distress for Vietnamese and Chinese caregivers. Identifying the caregiving and contextual variables associated with psychological distress is critical for tailoring interventions towards those who need the most help-in this case, possibly less educated White caregivers and more educated Asian American caregivers.


A Retrospective Assessment of the Continuous Health Care Provided to COVID-19 Patients Consulted Via Videoconference.
Telemedicine has been defined as a valuable tool in delivering care for COVID-19 patients. However, clinicians and policymakers should be convinced that traditional and new technological methods of clinical management may be equally effective. The purpose of this study was to generate some initial recommendations based on the clinical utility of videoconference consultation in forward triage and follow-up for COVID-19 patients. This retrospective cross-sectional study evaluated the medical records of 100 COVID-19 patients consulted using a videoconference program (Skype), from September 1, 2020, to February 3, 2021. The data were analyzed on demographic characteristics, disease history, the need for physical examination after videoconference consultation, pre-diagnostics and diagnostics, treatment decisions, number of videoconference consultation sessions in follow-up, duration of sessions, and final outcome. The male COVID-19 patients constituted 54% of the total sample. The median age was 51 (42-61) years. The median duration of the initial videoconference consultation session was 16 (12-21) minutes. Following the initial videoconference consultation session, 14 patients required follow-up with all face-to-face visits; the remaining patients were primarily followed with videoconference consultation sessions. For 25 patients, it was sufficient to provide only videoconference consultation sessions; they were not required to be in the hospital for physical examination or any subsequent investigation at all. A total of 14 patients were hospitalized. There was no statistically significant difference between the high-risk group and the other patients according to the components of the disease management process via videoconference consultation. Videoconference consultation enables a holistic assessment regardless of the patient's characteristics and allows for more time to be spent on each patient, particularly during the pandemic period without risk of contagion. It can be used as a forward triage and follow-up tool to identify patients in need of emergency hospitalization and continuous health care.


Hormonal status and age differentially affect tolerance to the disruptive effects of delta-9-tetrahydrocannabinol (Δ(9)-THC) on learning in female rats.
The effects of hormone status and age on the development of tolerance to Δ(9)-THC were assessed in sham-operated (intact) or ovariectomized (OVX) female rats that received either intraperitoneal saline or 5.6 mg/kg of Δ(9)-THC daily from postnatal day (PD) 75-180 (early adulthood onward) or PD 35-140 (adolescence onward). During this time, the four groups for each age (i.e., intact/saline, intact/THC, OVX/saline, and OVX/THC) were trained in a learning and performance procedure and dose-effect curves were established for Δ(9)-THC (0.56-56 mg/kg) and the cannabinoid type-1 receptor (CB1R) antagonist rimonabant (0.32-10 mg/kg). Despite the persistence of small rate-decreasing and error-increasing effects in intact and OVX females from both ages during chronic Δ(9)-THC, all of the Δ(9)-THC groups developed tolerance. However, the magnitude of tolerance, as well as the effect of hormone status, varied with the age at which chronic Δ(9)-THC was initiated. There was no evidence of dependence in any of the groups. Hippocampal protein expression of CB1R, AHA1 (a co-chaperone of CB1R) and HSP90β (a molecular chaperone modulated by AHA-1) was affected more by OVX than chronic Δ(9)-THC; striatal protein expression was not consistently affected by either manipulation. Hippocampal brain-derived neurotrophic factor expression varied with age, hormone status, and chronic treatment. Thus, hormonal status differentially affects the development of tolerance to the disruptive effects of delta-9-tetrahydrocannabinol (Δ(9)-THC) on learning and performance behavior in adolescent, but not adult, female rats. These factors and their interactions also differentially affect cannabinoid signaling proteins in the hippocampus and striatum, and ultimately, neural plasticity.


Study protocol for a randomized clinical trial to assess 7 versus 14-days of treatment for Pseudomonas aeruginosa bloodstream infections (SHORTEN-2 trial).
Research priorities in Antimicrobial Stewardship (AMS) have rapidly evolved in the last decade. The need for a more efficient use of antimicrobials have fueled plenty of studies to define the optimal duration for antibiotic treatments, and yet, there still are large areas of uncertainty in common clinical scenarios. Pseudomonas aeruginosa has been pointed as a priority for clinical research, but it has been unattended by most randomized trials tackling the effectiveness of short treatments. The study protocol of the SHORTEN-2 trial is presented as a practical example of new ways to approach common obstacles for clinical research in AMS. To determine whether a 7-day course of antibiotics is superior to 14-day schemes for treating bloodstream infections by P. aeruginosa (BSI-PA). A superiority, open-label, randomized controlled trial will be performed across 30 Spanish hospitals. Adult patients with uncomplicated BSI-PA will be randomized to receive a 7 versus 14-day course of any active antibiotic. The primary endpoint will be the probability for the 7-day group of achieving better outcomes than the control group, assessing altogether clinical effectiveness, severe adverse events, and antibiotic exposure through a DOOR/RADAR analysis. Main secondary endpoints include treatment failure, BSI-PA relapses, and mortality. A superiority design was set for the primary endpoint and non-inferiority for treatment failure, resulting in a sample size of 304 patients. SHORTEN-2 trial aligns with some of the priorities for clinical research in AMS. The implementation of several methodological innovations allowed overcoming common obstacles, like feasible sample sizes or measuring the clinical impact and unintended effects. EudraCt: 2021-003847-10; ClinicalTrials.gov: NCT05210439.


Accumulation of dietary and aqueous cadmium into the epidermal mucus of the discus fish Symphysodon sp.
The discus fish Symphysodon sp. is an Amazonian cichlid with a unusual form of parental care where fry obligately feed from parental mucus for the first few weeks of life. Here, we investigated the possible impact of environmental cadmium on this species, particularly with respect to mucus contamination. We exposed groups of fish to cadmium either through their food (400 mg kg(-1)) or through the water (3 μgl(-1)) for 4 weeks, and measured tissue concentrations and ATPase activities at weekly intervals. Cadmium significantly accumulated in all tissues (except for muscle) after 7 days, and tissue concentrations increased until the end of the experiment. Significant alterations in ATPase activities of intestine and kidney were observed at day 7 and 14, but no alterations in gill ATPase activities occurred. The epidermal mucus showed a high accumulation of cadmium from both exposures, but particularly from the diet, indicating that dietary cadmium can be transferred from gut to mucus. Combining this data with approximations of fry bite volumes and bite frequencies, we constructed daily estimates of the cadmium that could potentially be consumed by newly hatched fry feeding on this mucus. These calculations suggest that feeding fry might consume up to 11 μgg(-1)day(-1), and hence indicate that this species' dependency on parental mucus feeding of fry could make them particularly susceptible to cadmium contamination of their native habitat.


Sensitization of pancreatic cancer stem cells to gemcitabine by Chk1 inhibition.
Checkpoint kinase 1 (Chk1) inhibition sensitizes pancreatic cancer cells and tumors to gemcitabine. We hypothesized that Chk1 inhibition would sensitize pancreatic cancer stem cells to gemcitabine. We tested this hypothesis by using two patient-derived xenograft models (designated J and F) and the pancreatic cancer stem cell markers CD24, CD44, and ESA. We determined the percentage of marker-positive cells and their tumor-initiating capacity (by limiting dilution assays) after treatment with gemcitabine and the Chk1 inhibitor, AZD7762. We found that marker-positive cells were significantly reduced by the combination of gemcitabine and AZD7762. In addition, secondary tumor initiation was significantly delayed in response to primary tumor treatment with gemcitabine + AZD7762 compared with control, gemcitabine, or AZD7762 alone. Furthermore, for the same number of stem cells implanted from gemcitabine- versus gemcitabine + AZD7762-treated primary tumors, secondary tumor initiation at 10 weeks was 83% versus 43%, respectively. We also found that pS345 Chk1, which is a measure of DNA damage, was induced in marker-positive cells but not in the marker-negative cells. These data demonstrate that Chk1 inhibition in combination with gemcitabine reduces both the percentage and the tumor-initiating capacity of pancreatic cancer stem cells. Furthermore, the finding that the Chk1-mediated DNA damage response was greater in stem cells than in non-stem cells suggests that Chk1 inhibition may selectively sensitize pancreatic cancer stem cells to gemcitabine, thus making Chk1 a potential therapeutic target for improving pancreatic cancer therapy.


[The morphochemical characteristics of the brain in Wistar rats that differ by open-field motor activity].
By cytochemical and biochemical methods it is established that in rats with low motor activity the increased content of proteins of cytoplasma and nuclei of neurones of the sensorimotor cortex, caudate nucleus and n. accumbens, the increased activity of a number enzymes of oxidizing and protein metabolism in them are combined with a low activity of enzymes of mediator exchange. In the studied formations in rats with high motor activity an increased activity of synaptic and membrane forms of acetylcholinesterase appears at the same level of activity of cholineacetyltransferase in these subcortical formations and at high monoamineoxidase activity in cellular structures of the cortex and in subcortical formations. It is shown that in animals of the same line but differing by the behaviour in the open field, brain formations such as the sensorimotor cortex, caudate nucleus and n. accumbens have their own biochemical properties of the studied characteristics.


Human Microbiota and Breast Cancer-Is There Any Relevant Link?-A Literature Review and New Horizons Toward Personalised Medicine.
Breast cancer (BC) is the most common malignancy and the second cause of cancer-specific death in women from high-income countries. Recently, gut microbiota dysbiosis emerged as a key player that may directly and/or indirectly influence development, treatment, and prognosis of BC through diverse biological processes: host cell proliferation and death, immune system function, chronic inflammation, oncogenic signalling, hormonal and detoxification pathways. Gut colonisation occurs during the prenatal period and is later diversified over distinct phases throughout life. In newly diagnosed postmenopausal BC patients, an altered faecal microbiota composition has been observed compared with healthy controls. Particularly, β-glucuronidase bacteria seem to modulate the enterohepatic circulation of oestrogens and their resorption, increasing the risk of hormone-dependent BC. Moreover, active phytoestrogens, short-chain fatty acids, lithocholic acid, and cadaverine have been identified as bacterial metabolites influencing the risk and prognosis of BC. As in gut, links are also being made with local microbiota of tumoural and healthy breast tissues. In breast microbiota, different microbial signatures have been reported, with distinct patterns per stage and biological subtype. Total bacterial DNA load was lower in tumour tissue and advanced-stage BC when compared with healthy tissue and early stage BC, respectively. Hypothetically, these findings reflect local dysbiosis, potentially creating an environment that favours breast tumour carcinogenesis (oncogenic trigger), or the natural selection of microorganisms adapted to a specific microenvironment. In this review, we discuss the origin, composition, and dynamic evolution of human microbiota, the links between gut/breast microbiota and BC, and explore the potential implications of metabolomics and pharmacomicrobiomics that might impact BC development and treatment choices toward a more personalised medicine. Finally, we put in perspective the potential limitations and biases regarding the current microbiota research and provide new horizons for stronger accurate translational and clinical studies that are needed to better elucidate the complex network of interactions between host, microorganisms, and drugs in the field of BC.


Cytotoxic reaction and TNF-alpha response of macrophages to polyurethane particles.
Their unique mechanical and biological properties make polyurethanes (PUs) ideal materials for many implantable devices. However, uncertain long-term biostability in the human physiological environment limits their extensive clinical applications. Chronic inflammatory response associated with macrophage activation has been suggested as a prime factor; although the mechanism of macrophage activation in response to biomaterial surfaces and debris is still unknown. The overall objective of this work was to study the response of macrophages to PU materials in vitro by measuring cell viability and activity. The studies were carried out using phagocytozable-size PU particles from three types of commercially-available PUs: Pellethane 2363 80ABA (PL); Tecothane TT2065 (TC65); and Tecothane TT2085 (TC85). These polymers posess the same generic composition but differ in the length of hard and soft segments, as revealed by the FTIR and NMR studies. The results showed that PU particles affected both viability and activity of J774 macrophages. The percentage of mortality ranged from 1 to 15% with 10-100 microg ml(-1) of particles after 24 and 48 h incubation. These three types of particles induced different mortality on the macrophages. Specifically, the mortality with PL particles was 1-4% (p > 0.05), while the mortality with TC85 particles was 2-10% (p < 0.05) and 4-15% with TC65 (p < 0.05). Conversely, these particles also affected cell proliferation. Cell numbers increased by 132 and 167% after 24 and 48 h incubation, respectively, without particles, whereas the cell numbers increased only 46 and 78% with TC65, 66 and 105% with TC85, and 67 and 110% with PL in the presence of 100 microg ml(-1) of particles for the respective incubation times. PU particles also increased TNF-alpha release from macrophage. After having been incubated for 24 h with 100 microg ml(-1) particles of TC65, TC85, and PL, macrophages release TNF-alpha 7.4, 5.2, and 4.1 times more than the control. In conclusion, PU particles had cytotoxic effects on J774 macrophage at high concentrations. The order of macrophage response for three types of particles was TC65 > TC85 > PL. PU particles' effect on macrophage viability and activity depends on the concentration of particles and their chemical composition, especially on the ratio of hard to soft segments.


Lipid-altering efficacy and safety of ezetimibe/simvastatin versus atorvastatin in patients with hypercholesterolemia and the metabolic syndrome (from the VYMET study).
Patients with the metabolic syndrome are at an increased risk of cardiovascular disease and might require intensive lipid therapy. Many patients remain at the starting dose of lipid therapy and might not be titrated up to a higher dose. The present double-blind, randomized, 6-week study assessed the lipid-lowering efficacy of ezetimibe/simvastatin 10/20 mg versus atorvastatin 10 or 20 mg, and ezetimibe/simvastatin 10/40 mg versus atorvastatin 40 mg in 1,128 patients with hypercholesterolemia and the metabolic syndrome. The primary end point was the percentage of change from baseline in low-density lipoprotein (LDL) cholesterol. Additional end points included changes in other lipids, lipoprotein ratios, high-sensitivity C-reactive protein, and attainment of prespecified lipid levels. Significantly greater improvements in the levels of LDL cholesterol, non-high-density lipoprotein cholesterol, apolipoprotein B, and lipid/lipoprotein ratios resulted with ezetimibe/simvastatin compared with atorvastatin at all specified dose comparisons (p <0.001). The attainment of prespecified LDL cholesterol and non-high-density lipoprotein cholesterol levels was also significantly greater with ezetimibe/simvastatin than with atorvastatin at all dose comparisons (p <0.05). High-density lipoprotein cholesterol increases were significantly greater with ezetimibe/simvastatin 10/20 mg than with atorvastatin 10 mg (p <0.05) and ezetimibe/simvastatin 10/40 mg than with atorvastatin 40 mg (p <0.01). The changes in triglycerides, very low-density lipoprotein cholesterol, and high-sensitivity C-reactive protein were similar for both treatments. The incidence of liver, muscle, and gastrointestinal-, hepatitis- and allergic reaction/rash-related adverse events were low and generally similar to those in previous studies of ezetimibe/simvastatin and/or atorvastatin. In conclusion, ezetimibe/simvastatin was more likely to result in lipid treatment end points than atorvastatin and was generally well tolerated at the doses compared in our patients.


The power and potential of doxorubicin-DNA adducts.
Doxorubicin (trade name Adriamycin) is a widely used anticancer agent which exhibits good activity against a wide range of tumors. Although the major mode of action appears to be normally as a topoisomerase II poison, it also exhibits a number of other cellular responses, one of which is the ability to form adducts with DNA. For adduct formation doxorubicin must react with cellular formaldehyde to form an activated Schiff base which is then able to form an aminal (N-C-N) linkage to the exocyclic amino group of guanine residues. The mono-adducts form primarily at G of 5'-GCN-3' sequences where the chromophore of the drug is intercalated between the C and N base pair. The structure of the adducts has have been well defined by 2D NMR, mass spectrometry and X-ray crystallography. The formation of these anthracycline adducts in cells grown in culture has been unequivocally demonstrated. The source of formaldehyde in cells can be endogenous, provided by coadministration of prodrugs that release formaldehyde or by prior complexation of anthracyclines with formaldehyde. Since the adducts appear to be more cytotoxic than doxorubicin alone, and also less susceptible to drug-efflux forms of resistance, they offer new approaches to improving the anticancer activity of the anthracyclines.


Home Care of Children With Diarrhea in Bangui's Therapeutic Landscape (Central African Republic).
We explore how the therapeutic landscape of Bangui, capital of the Central African Republic, shaped and reflected caregivers' home care of childhood diarrhea. Using interviews, group discussions, and participant observation, we found that caregivers described several categories of diarrhea, but in practice, adopted an experimental approach to home diarrheal care. Many managed incertitude by initially dosing children with street medicines and herbal infusions; they delayed seeking professional medical care to avoid expenses, observed their children's symptoms, consulted social networks, and used therapeutic foods. The logics underpinning these practices emerged from caregivers' conviction that diarrhea necessitated medical therapy and restricted choices within Bangui's therapeutic landscape, a consequence of lengthy political economic crisis. This crisis impoverished Bangui populations, eroded formal health care, and sharpened religious differences, discouraging care seeking from traditional healers. Analyses of therapeutic landscapes illuminate why caregivers embraced specific practices and logics and should guide the elaboration of more effective public health interventions.


Risk moderates the outcome of relationship education: a randomized controlled trial.
To test whether the effects of relationship education programs generalize across couples regardless of their baseline levels of risk for relationship distress, or whether intervention effects vary systematically as a function of risk. The former result would support primary prevention models; the latter result would support a shift toward secondary prevention strategies. Engaged and newlywed couples (N = 130) were randomized into 1 of 3 relationship education programs. Individual and relational risk factors assessed at baseline were tested as moderators of 3-year changes in relationship satisfaction, overall and in each of the 3 treatment conditions. Treatment effects varied as a function of risk, and more so with variables capturing relational risk factors than individual risk factors. High-risk couples (e.g., couples with lower levels of baseline commitment and satisfaction) tended to decline less rapidly in satisfaction than low-risk couples following treatment. Couples with acute concerns at baseline, including higher levels of physical aggression and alcohol use, benefitted less from intervention than couples without these concerns. Comparisons across treatment conditions indicate that couples with relatively high baseline satisfaction and commitment scores declined faster in satisfaction when assigned to an intensive skill-based intervention, as compared with a low-intensity intervention. Outcomes of skill-based relationship education differ depending on premarital risk factors. Efficient identification of couples at risk for adverse relationship outcomes is needed to refine future prevention efforts, and deploying prevention resources specifically to at-risk populations may be the most effective strategy for strengthening couples and families.


Determination of absolute configuration of the phosphonic acid moiety of fosfazinomycins.
Fosfazinomycins A and B produced by Streptomyces lavendofoliae share the same phosphonate moiety with one chiral centre of unknown configuration which was determined by synthesising both enantiomers of 2-hydroxy-2-phosphonoacetic acid methyl ester. A chiral cyclic phosphite was reacted with methyl glyoxylate in a Pudovik reaction to give a pair of diastereomeric α-hydroxyphosphonates, which were separated by HPLC. The configurations at C-2 were assigned on the basis of single crystal X-ray structure analysis. Deprotection of these diastereomers furnished the enantiomeric α-hydroxyphosphonic acids, of which the (S)-configured had the same sign of optical rotation as the phosphonic acid moiety of the two fosfazinomycins.


Diabetogenic effect of gluten in Wistar albino rats: a preliminary preclinical screening.
Gluten-related disease affects less than 1% population and is not considered of relevance at the public health level. However, the consumption of a gluten-free diet has been most commonly adopted as a special diet worldwide in the recent past. In the present study, we investigated the association of gluten intake and diabetes in Wistar albino rats. Thirty adult Wistar rats were randomly divided into five groups: control, diabetic, and test treated with pure gluten (100, 200 and 400 mg/kg body weight). Diabetes was induced in rats by intraperitoneal injection of Streptozotocin (65 mg/kg) after a dose of nicotinamide (110 mg/kg). Body weight, fasting blood glucose levels, postprandial blood glucose levels and histopathology of the pancreas were compared. Fasting blood glucose levels and postprandial blood glucose were significantly higher in diabetes animals but there were no significant changes in gluten treated groups. Other parameters were not significantly changed among different groups. Gluten at doses 100 mg/kg, 200 mg/kg and 400 mg/kg is not a diabetogenic diet and hence it needs not be excluded from diet for the prevention and management of Type 2 diabetes mellitus.


Deep learning and targeted metabolomics-based monitoring of chewing insects in tea plants and screening defense compounds.
Tea is an important cash crop that is often consumed by chewing pests, resulting in reduced yields and economic losses. It is important to establish a method to quickly identify the degree of damage to tea plants caused by leaf-eating insects and screen green control compounds. This study was performed through the combination of deep learning and targeted metabolomics, in vitro feeding experiment, enzymic analysis and transient genetic transformation. A small target damage detection model based on YOLOv5 with Transformer Prediction Head (TPH-YOLOv5) algorithm for the tea canopy level was established. Orthogonal partial least squares (OPLS) was used to analyze the correlation between the degree of damage and the phenolic metabolites. A potential defensive compound, (-)-epicatechin-3-O-caffeoate (EC-CA), was screened. In vitro feeding experiments showed that compared with EC and epicatechin gallate, Ectropis grisescens exhibited more significant antifeeding against EC-CA. In vitro enzymatic experiments showed that the hydroxycinnamoyl transferase (CsHCTs) recombinant protein has substrate promiscuity and can catalyze the synthesis of EC-CA. Transient overexpression of CsHCTs in tea leaves effectively reduced the degree of damage to tea leaves. This study provides important reference values and application prospects for the effective monitoring of pests in tea gardens and screening of green chemical control substances.


Loss in the Intrinsic Quality and the Antioxidant Activity of Sunflower (Helianthus annuus L.) Oil during an Industrial Refining Process.
Minor compounds in vegetable oils are of health interest due to their powerful biological antioxidant properties. In order to extend the shelf life of sunflower oil, it is generally subjected to a refining process that can affect these desirable compounds. The main purpose of this study was to determine the effect of this chemical/physical refining process on selected minor components of sunflower oil in order to establish the nutritional quality and health properties of the oil. The oxidative stability, contents of fatty acids, tocopherols, phytosterols, reducing capacity, β-carotene, chlorophyll, and squalene were studied during six refining steps. Quantitative data showed the evolution of oil quality according to its degree of refinement. The results showed a significant decrease for all of the minor compounds analyzed, with losses in carotenoids of 98.6%, 8.5% in tocopherols, 19.5% in phytosterols and 45.0% in squalene. The highest reductions were recorded for the compounds that alter the most the visual aspects of the oil (waxes, carotenoids and chlorophylls) whereas reduction was limited for the compounds with no impact on the organoleptic quality. The losses in the compounds of health interest should be minimized by improving the refining processes and/or having a greater content of those molecules in crude oil by breeding new performing varieties.


Down-regulated TAB1 suppresses the replication of Coxsackievirus B5 via activating the NF-κB pathways through interaction with viral 3D polymerase.
Coxsackievirus Group B type 5 (CVB5), an important pathogen of hand-foot-mouth disease, is also associated with neurological complications and poses a public health threat to young infants. Among the CVB5 proteins, the nonstructural protein 3D, known as the Enteroviral RNA-dependent RNA polymerase, is mainly involved in viral genome replication and transcription. In this study, we performed immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify host proteins that interacted with CVB5 3D polymerase. A total of 116 differentially expressed proteins were obtained. Gene Ontology analysis identified that the proteins were involved in cell development and cell adhesion, distributed in the desmosome and envelope, and participated in GTPase binding. Kyoto Encyclopedia of Genes and Genomes analysis further revealed they participated in nerve diseases, such as Parkinson disease. Among them, 35 proteins were significantly differentially expressed and the cellular protein TGF-BATA-activated kinase1 binding protein 1 (TAB1) was found to be specifically interacting with the 3D polymerase. 3D polymerase facilitated the entry of TAB1 into the nucleus and down-regulated TAB1 expression via the lysosomal pathway. In addition, TAB1 inhibited CVB5 replication via inducing inflammatory factors and activated the NF-κB pathway through IκBα phosphorylation. Moreover, the 90-96aa domain of TAB1 was an important structure for the function. Collectively, our findings demonstrate the mechanism by which cellular TAB1 inhibits the CVB5 replication via activation of the host innate immune response, providing a novel insight into the virus-host innate immunity.


Effect of competitive interactions on the structure and properties of blends prepared from an industrial lignosulfonate polymer.
To explore the possibility of applying lignin in practice, an industrial lignosulfonate (0-50 vol%) was blended with four ionomers. The concentrations of carboxyl and carboxylate groups were systematically varied in the ethylene-acrylic acid copolymers to study the competition of hydrogen and ionic bonds forming between the components. The mechanical properties of the blends were determined by tensile testing. The structure was investigated by scanning electron microscopy, while deformation and failure processes were studied by acoustic emission measurements and microscopy. Interfacial interactions were quantitatively characterized by analyzing local deformation processes and by evaluating the composition dependence of the tensile strength using appropriate models. Molecular dynamics simulations indicated that carboxylate groups preferably form clusters in the ionomer phase, consequently, the increasing degree of neutralization results in ionomers with more and more self-interactions of components deteriorating ionomer-lignin interactions. The novel combination of experiments, modeling, and simulation was done for the first time on such materials, and it pointed out that the role of hydrogen bonds is more critical in determining blend properties. Blends can be prepared for practical applications with a good combination of stiffness (0.8 GPa), tensile strength (22 MPa), and elongation-at-break (25 %) at 30 vol% lignosulfonate content and 33 % neutralization.


[Blocking action of calmidazolium and chlorpromazine on the potassium outward current which depends on intracellular calcium ions].
Effects of calmidazolium (R 24571) and chlorpromazine on the delayed potassium outward current in the somatic membrane were studied on nonidentified intracellularly perfused neurons of the snail Helix pomatia. Extracellular application of these substance evoked depression of the outward current. Inhibition of IK occurs at concentrations of calmodulin inhibitors 10(-9)-10(-8) mol/l. These agents inhibit primarily a component of the potassium current depending on the intracellular Ca2+ ions (IK(Ca in)). The inhibitory effect of these drugs can be explained by calmodulin-like structures of the receptor for intracellular calcium, providing modulation of IK(Ca in).


Differential effect of cimetidine on drug oxidation (antipyrine and diazepam) vs. conjugation (acetaminophen and lorazepam): prevention of acetaminophen toxicity by cimetidine.
Fourteen healthy volunteers received a single i.v. dose of antipyrine (1.2 g) on two occasions, once before and once during cimetidine treatment (300 mg every 6 hr). In a similar manner, 8 subjects received diazepam (10 mg), 11 subjects received acetaminophen (650 mg) and 8 subjects received lorazepam (2 mg), all by the i.v. route, once before and once during cimetidine coadministration. Pharmacokinetic analysis indicated increased antipyrine elimination T1/2 during cimetidine treatment (16.7 vs. 10.9 hr; P less than .001) on the basis of decreased total metabolic clearance (0.46 vs. 0.72 ml/min/kg; P less than .001). Likewise, diazepam T1/2 was increased (58 vs. 39 hr; P less than .01) during cimetidine treatment due to decreased total metabolic clearance (0.42 vs. 0.30 ml/min/kg; P less than .01). In contrast, cimetidine did not alter T1/2 or the clearance of lorazepam (T1/2, 16.8 vs. 15.3 hr; clearance, 1.03 vs 1.07 ml/min/kg) or acetaminophen (T1/2, 2.66 vs. 2.60 hr; clearance, 4.8 vs. 4.5 ml/min/kg), both drugs which undergo conjugative biotransformation. In an animal model used to assess the effect of cimetidine on acetaminophen toxicity, the LD50 of acetaminophen alone in Charles River CD-1 mice was 480 mg/kg (95% confidence interval: 436-528 mg/kg). With simultaneous 75 mg/kg of cimetidine treatment, the LD50 for acetaminophen was significantly increased (P less than .05) to 1020 mg/kg (95% confidence interval: 962-1081 mg/kg). Thus, cimetidine slows the metabolic clearance of antipyrine and diazepam, drugs biotransformed by hepatic oxidation, but does not alter the kinetics of acetaminophen or lorazepam, both metabolized by conjugation. Cimetidine may decrease the toxicity of high-dose acetaminophen by preventing formation of the hepatotoxic oxidative metabolites, although having no effect on conjugation of acetaminophen which yields nontoxic metabolites that are subsequently cleared from the body.


Using mice to decipher the molecular genetics of brain tumors.
The past decade has dramatically increased our knowledge of genetic and molecular alterations in human central nervous system tumors. Important as these alterations are for the molecular classification of tumors, their actual roles in tumorigenesis and tumor progression have long remained obscure. Lately, several mouse brain tumor models have been developed that use different gene modification strategies to replicate mutations seen in the human counterpart. These genetic models will allow discrimination between mutations that are causally related to tumor formation and mutations that are a result of tumor progression. These models also provide histologically and genetically accurate models for preclinical testing and will perhaps help us identify novel targets for therapies aimed at the mechanistic cause of the disease. We present here a review of current models, with a focus on gliomas and medulloblastomas.


Bioconcentration of the intense sweetener sucralose in a multitrophic battery of aquatic organisms.
Reports of the intense (artificial) sweetener sucralose (1,6-dichloro-1,6-dideoxy-β-D-fructo-furanosyl 4-chloro-4-deoxy-α-D-galactopyranoside) in various environmental compartments have led to speculations about biological effects in nontarget species living in areas receiving discharges from anthropogenic activities. We have, as the first step in the risk assessment of this compound, conducted bioaccumulation studies in the freshwater alga Pseudokirchneriella subcapitata, the crustacean Daphnia magna, and zebrafish (Danio rerio). The freshwater algae and the daphnid tests were performed using a 48-h static exposure system, whereas the zebrafish test was performed using a 48-h semi static exposure system followed by 48 h flow-through of clean water for the depuration phase. All three studies were conducted with two exposure concentrations (10 and 100 mg/L), and the concentrations of sucralose in water and biota were verified by liquid chromatography/mass spectrometry. The studies showed that uptake of sucralose was assumed to achieve a steady state within the first 48 h, and the bioconcentration factor at the assumed steady state (BCF(SS) ) was calculated to be less than 1 for algae and between 1.6 to 2.2 for the daphnids. The fish BCF(SS), assumed to occur between 24 to 48 hours, were calculated to be less than 1 for both concentrations tested. A first-order one-compartment (uptake phase) and a first-order two-compartment (elimination phase) model characterized the uptake and depuration kinetics in zebrafish (k(1)=0.027-0.038/h and k(2)=0.206-0.222/h, t(95)=13.5 to 14.6 h, t(50)=3.1 to 3.3 h, and BCF(kinetic)=0.4 to 0.9). The current study shows that sucralose does not bioaccumulate in aquatic organisms from different tiers of the food web, and that the BCF's obtained were considerably lower than the criteria set to identify persistent, bioaccumulative, and toxic substances (i.e., BCF ≥ 2,000).


Acoustic shock.
Acoustic shock is a recently recognised clinical entity: following an abrupt, intense and unanticipated acoustic stimulus, usually delivered by a telephone handset or headset, some individuals report a symptom cluster that includes otalgia, altered hearing, aural fullness, imbalance, tinnitus, dislike or even fear of loud noises, and anxiety and/or depression. Symptoms start shortly after the triggering acoustic incident and can be short-lived or can last for a considerable time. If persistent, the condition can lead to significant disability. Proposed mechanisms include involvement of the tensor tympani muscle, hyperexcitability of central auditory pathways, and a precursive state of raised anxiety or arousal. A formal treatment programme has not yet been proposed, but the potential utility of modern therapeutic techniques for tinnitus and hyperacusis are considered. Given the large number of UK residents working in telephone call centres, this condition is of considerable clinical importance.


The synaptic cell adhesion molecule, SynCAM1, mediates astrocyte-to-astrocyte and astrocyte-to-GnRH neuron adhesiveness in the mouse hypothalamus.
We previously identified synaptic cell adhesion molecule 1 (SynCAM1) as a component of a genetic network involved in the hypothalamic control of female puberty. Although it is well established that SynCAM1 is a synaptic adhesion molecule, its contribution to hypothalamic function is unknown. Here we show that, in addition to the expected neuronal localization illustrated by its presence in GnRH neurons, SynCAM1 is expressed in hypothalamic astrocytes. Cell adhesion assays indicated that SynCAM is recognized by both GnRH neurons and astrocytes as an adhesive partner and promotes cell-cell adhesiveness via homophilic, extracellular domain-mediated interactions. Alternative splicing of the SynCAM1 primary mRNA transcript yields four mRNAs encoding membrane-spanning SynCAM1 isoforms. Variants 1 and 4 are predicted to be both N and O glycosylated. Hypothalamic astrocytes and GnRH-producing GT1-7 cells express mainly isoform 4 mRNA, and sequential N- and O-deglycosylation of proteins extracted from these cells yields progressively smaller SynCAM1 species, indicating that isoform 4 is the predominant SynCAM1 variant expressed in astrocytes and GT1-7 cells. Neither cell type expresses the products of two other SynCAM genes (SynCAM2 and SynCAM3), suggesting that SynCAM-mediated astrocyte-astrocyte and astrocyte-GnRH neuron adhesiveness is mostly mediated by SynCAM1 homophilic interactions. When erbB4 receptor function is disrupted in astrocytes, via transgenic expression of a dominant-negative erbB4 receptor form, SynCAM1-mediated adhesiveness is severely compromised. Conversely, SynCAM1 adhesive behavior is rapidly, but transiently, enhanced in astrocytes by ligand-dependent activation of erbB4 receptors, suggesting that erbB4-mediated events affecting SynCAM1 function contribute to regulate astrocyte adhesive communication.


Patient considerations in the management of menopausal symptoms: role of conjugated estrogens with bazedoxifene.
Menopausal symptoms (eg, hot flushes and vaginal symptoms) are common, often bothersome, and can adversely impact women's sexual functioning, relationships, and quality of life. Estrogen-progestin therapy was previously considered the standard care for hormone therapy (HT) for managing these symptoms in nonhysterectomized women, but has a number of safety and tolerability concerns (eg, breast cancer, stroke, pulmonary embolism, breast pain/tenderness, and vaginal bleeding) and its use has declined dramatically in the past decade since the release of the Women's Health Initiative trial results. Conjugated estrogens paired with bazedoxifene (CE/BZA) represent a newer progestin-free alternative to traditional HT for nonhysterectomized women. CE/BZA has demonstrated efficacy in reducing the frequency and severity of vasomotor symptoms and preventing loss of bone mineral density in postmenopausal women. CE/BZA provides an acceptable level of protection against endometrial hyperplasia and does not increase mammographic breast density. Compared with traditional estrogen-progestin therapy, it is associated with lower rates of breast pain/tenderness and vaginal bleeding. Patient-reported outcomes indicate that CE/BZA improves menopause-specific quality of life, sleep, some measures of sexual function (especially ease of lubrication), and treatment satisfaction. This review looks at the rationale for selection and combination of CE with BZA at the dose ratio in the approved product and provides a detailed look at the efficacy, safety, tolerability, and patient-reported outcomes from the five Phase III trials. Patient considerations in the choice between CE/BZA and traditional HT (eg, tolerability, individual symptoms, and preferences for route of administration) are also considered.


Humanin gene expression in fibroblast of Down syndrome subjects.
Down syndrome (DS) is characterized by trisomy of chromosome 21 and peculiar phenotype. Humanin (HN) is a mitochondrial short 24-residue polypeptide whit anti-apoptotic and neuroprotective effects. In this study we evaluated HN protein expression and HN mRNA levels in cultured fibroblasts from DS patients and normal controls. Our results obtained by immunocytochemistry, western-blot and qRT-PCR analysis show a significant HN up-regulation in DS patients. These results confirm previous studies and suggest a role for HN may in the DS phenotype.


Increased Na+ intake during gestation in rats is associated with enhanced vascular reactivity and alterations of K+ and Ca2+ function.
Gestation is associated with decreased blood pressure and resistance to the effects of vasoconstrictor agents. A recent study showed that pregnant rats, on increased sodium intake, present physiological changes that resemble those observed in preeclampsia. We investigated the effects of sodium supplementation on reactivity and on potassium and Ca(2+) channel activity in blood vessels during gestation. Sodium supplements, 0.9% or 1.8% NaCl as drinking water, were given to nonpregnant and pregnant rats for 7 days (last week of gestation). Reactivity to phenylephrine (PE), KCl, arginine vasopressin (AVP), and tetraethylammonium (TEA) was measured in aortic rings under modulation of potassium and calcium channels. TEA, a nonselective K(+) channel inhibitor, induced concentration-dependent responses in aortic rings from nonpregnant but not in those from pregnant rats. The response to TEA was restored in rings from pregnant rats after preincubation with 10 mmol/l KCl. Sodium supplementation did not affect the response to TEA in the aortas of pregnant animals. After sodium supplementation, maximum responses to PE and AVP were decreased and increased in aortic rings from nonpregnant and pregnant rats, respectively. Cromakalim (an ATP-sensitive K(+) channel activator)-induced inhibition of the responses to the three vasoconstrictors was more striking in aorta from nonpregnant than pregnant rats on regular diet, whereas it produced similar inhibition in tissues from both groups of animals on 0.9% and 1.8% NaCl. NS-1619 (a Ca(2+)-sensitive K(+) activator) elicited heightened effects in the aortas of pregnant animals receiving 0.9% NaCl supplementation. Nifedipine (a Ca(2+) channel blocker) caused greater inhibition of the contractile responses in tissues from nonpregnant rats on regular diet, and its action was increased in pregnant rats on sodium-supplemented diets. These data demonstrate that augmented sodium intake during gestation in the rat is linked with the reversal of gestational-associated resistance to vasopressors and indicate that this is an experimental model showing some features of gestational hypertension.


Ashwagandha in brain disorders: A review of recent developments.
Withania somnifera (Family: Solanaceae), commonly known as Ashwagandha or Indian ginseng is distributed widely in India, Nepal, China and Yemen. The roots of plant consist of active phytoconstituents mainly withanolides, alkaloids and sitoindosides and are conventionally used for the treatment of multiple brain disorders. This review aims to critically assess and summarize the current state and implication of Ashwagandha in brain disorders. We have mainly focussed on the reported neuroactive phytoconstituents, available marketed products, pharmacological studies, mechanism of action and recent patents published related to neuroprotective effects of Ashwagandha in brain disorders. All the information and data was collected on Ashwagandha using keywords "Ashwagandha" along with "Phytoconstituents", "Ayurvedic, Unani and Homeopathy marketed formulation", "Brain disorders", "Mechanism" and "Patents". Following sources were searched for data collection: electronic scientific databases such as Science Direct, Google Scholar, Elsevier, PubMed, Wiley On-line Library, Taylor and Francis, Springer; books such as AYUSH Pharmacopoeia; authentic textbooks and formularies. Identified neuroprotective phytoconstituents of Ashwagandha are sitoindosides VII-X, withaferin A, withanosides IV, withanols, withanolide A, withanolide B, anaferine, beta-sitosterol, withanolide D with key pharmacological effects in brain disorders mainly anxiety, Alzheimer's, Parkinson's, Schizophrenia, Huntington's disease, dyslexia, depression, autism, addiction, amyotrophic lateral sclerosis, attention deficit hyperactivity disorder and bipolar disorders. The literature survey does not highlight any toxic effects of Ashwagandha. Further, multiple available marketed products and patents recognized its beneficial role in various brain disorders; however, very few data is available on mechanistic pathway and clinical studies of Ashwagandha for various brain disorders is scarce and not promising. The review concludes the results of recent studies on Ashwagandha suggesting its extensive potential as neuroprotective in various brain disorders as supported by preclinical studies, clinical trials and published patents. However vague understanding of the mechanistic pathways involved in imparting the neuroprotective effect of Ashwagandha warrants further study to promote it as a promising drug candidate.


Arginine need of heavy broiler males: applying the ideal protein concept.
An experiment was conducted to determine arginine need of male broilers between the ages of 42 to 56 d, in conjunction with dietary protein approaching a previously advocated ideal amino acid pattern. Ross x Ross 308 chicks were reared in floor pens (32 pens with 35 birds each) of an open-sided house on common feeds until 42 d of age. From 42 to 56 d of age, birds were fed a corn-soybean meal diet (17% CP, 3,250 kcal/kg ME, and 0.85% lysine) having basal arginine at 0.80%, and then progressive additions of 0.15% were made until 1.25% was reached to form the dietary treatments. Final body weight together with body weight gain and feed conversion through the 42-to-56-d experimental period were optimized at 0.98% arginine. Weight of the chilled carcass was optimized at 1.00% arginine, whereas depot fat that had been removed from the abdominal cavity continued to decrease to the highest level of supplementation. Additional total arginine to 1.05% was needed to maximize weight recovery of fillets and total breast meat. An arginine requirement for nutritional purposes approximating 1.00% as advocated by NRC (1994) is in general agreement with present results for live production and meat yield; however, carcass incidence of skin scratch infections and parts defects from processing stresses continually responded until the highest level to suggest that additional amounts would be needed for immunological and connective tissue challenges.


Angiotensin II Blockade and Total Cardiovascular Risk : Beyond Blood Pressure Reduction.
Hypertension is one of the world's largest public health problems and it is both a disease and a risk factor for cardiovascular disease (CVD). The heart, the endothelium and the kidneys are the target organs of hypertension. Recently, several antihypertensive drugs have been introduced to the market; therefore, the choice is mainly determined by the patients' features. In particular, ACE inhibitors and angiotensin II type 1 receptor antagonists (angiotensin receptor blockers [ARBs]) demonstrate a larger reduction in cardiovascular risk than other antihypertensive treatments because of the existence of blood pressure-independent effects. In fact, the angiotensin II pathway plays a major role in metabolic, haemodynamic and endothelial homeostasis. For these reasons, ACE inhibitors and ARBs have primary indications in patients with obesity, hypercholesterolaemia and diabetes mellitus because of their favourable metabolic properties. Furthermore, several large trials have demonstrated that they have favourable effects also in patients with left ventricular dysfunction or systolic heart failure, as well as other forms of heart disease. Drugs affecting the angiotensin II pathway may reduce endothelial dysfunction through several mechanisms including reduction of vascular permeability and oxidative stress. Another important effect of these drugs is neuroprotection. This is an important effect because in the near future, due to an aging population, an important goal for optimal antihypertensive treatment will be the prevention of cognitive decline. ACE inhibitors and ARBs are very important drugs in the modern management of the total cardiovascular risk in hypertensive patients.


Stochastic simplified modelling of abrasive waterjet footprints.
Abrasive micro-waterjet processing is a non-conventional machining method that can be used to manufacture complex shapes in difficult-to-cut materials. Predicting the effect of the jet on the surface for a given set of machine parameters is a key element of controlling the process. However, the noise of the process is significant, making it difficult to design reliable jet-path strategies that produce good quality parts via controlled-depth milling. The process is highly unstable and has a strong random component that can affect the quality of the workpiece, especially in the case of controlled-depth milling. This study describes a method to predict the variability of the jet footprint for different jet feed speeds. A stochastic partial differential equation is used to describe the etched surface as the jet is moved over it, assuming that the erosion process can be divided into two main components: a deterministic part that corresponds to the average erosion of the jet and a stochastic part that accounts for the noise generated at different stages of the process. The model predicts the variability of the trench profiles to within less than 8%. These advances could enable abrasive micro-waterjet technology to be a suitable technology for controlled-depth milling.


Effect of specific neutrophil elastase inhibitor on ischemia/reperfusion injury in rat liver transplantation.
Activated neutrophils have been implicated as playing an important role in ischemia/reperfusion injury of the liver by releasing toxic mediators such as oxygen free radicals and elastases. In the present study, we evaluated the effect of a novel, specific neutrophil elastase inhibitor (ONO-5046) on cold-ischemia/reperfusion injury of the liver allograft in rodents. Livers from male Lewis rats were procured and stored cold (4 degrees C) in lactated Ringer's solution and transplanted orthotopically. Recipients were divided into three groups: Vehicle group, 5-h preservation and vehicle (n = 8); ONO-5046 group, 5-h preservation and administration of ONO-5046 (n = 8); and Control group, minimum preservation only (n = 8). Bile output after reperfusion was significantly larger in the ONO-5046 group compared to the Vehicle group (P < 0.05 or less). Sinusoidal endothelial cell function represented by the serum hyaluronic acid concentration at 120 min after reperfusion of the ONO-5046 group was significantly lower than that in the Vehicle group (17.0 +/- 7.9 vs 36.2 +/- 14.9 ng/ml, P < 0.05), whereas serum transaminase levels 120 min after reperfusion were comparable between the two groups. Liver tissue energy charge 120 min after reperfusion was significantly better in the ONO-5046 group compared to the Vehicle group (P < 0.05). Furthermore, the number of neutrophils infiltrating the allograft after reperfusion was significantly depressed in the ONO-5046 group compared to the Vehicle group (P < 0. 02). These data suggest that the neutrophil elastase might cause liver damage early after reperfusion in cold-stored liver, which can be ameliorated by the administration of a specific neutrophil elastase inhibitor, ONO-5046.


Mechanisms of sperm chemotaxis.
Sperm are attracted by chemical factors that are released by the egg-a process called chemotaxis. Most of our knowledge on sperm chemotaxis originates from the study of marine invertebrates. In recent years, the main features of the chemotactic signaling pathway and the swimming behavior evoked by chemoattractants have been elucidated in sea urchins. In contrast, our understanding of mammalian sperm chemotaxis is still rudimentary and subject to an ongoing debate. In this review, we raise new questions and discuss current concepts of sperm chemotaxis. Finally, we highlight commonalities and differences of sensory signaling in sperm, photoreceptors, and olfactory neurons.


Increased susceptibility to ATP via alteration of P2X receptor function in dystrophic mdx mouse muscle cells.
Pathological cellular hallmarks of Duchenne muscular dystrophy (DMD) include, among others, abnormal calcium homeostasis. Changes in the expression of specific receptors for extracellular ATP in dystrophic muscle have been recently documented: here, we demonstrate that at the earliest, myoblast stage of developing dystrophic muscle a purinergic dystrophic phenotype arises. In myoblasts of a dystrophin-negative muscle cell line established from the mdx mouse model of DMD but not in normal myoblasts, exposure to extracellular ATP triggered a strong increase in cytoplasmic Ca2+ concentrations. Influx of extracellular Ca2+ was stimulated by ATP and BzATP and inhibited by zinc, Coomassie Brilliant Blue-G, and KN-62, demonstrating activation of P2X7 receptors. Significant expression of P2X4 and P2X7 proteins was immunodetected in dystrophic myoblasts. Therefore, full-length dystrophin appears, surprisingly, to play an important role in myoblasts in controlling responses to ATP. Our results suggest that altered function of P2X receptors may be an important contributor to pathogenic Ca2+ entry in dystrophic mouse muscle and may have implications for the pathogenesis of muscular dystrophies. Treatments aiming at inhibition of specific ATP receptors could be of a potential therapeutic benefit.


Bioinspired glycosaminoglycan hydrogels via click chemistry for 3D dynamic cell encapsulation.
Cell encapsulation within 3D hydrogels is an attractive approach to develop effective cell-based therapies. However, little is known about how cells respond to the dynamic microenvironment resulting from hydrogel gelation-based cell encapsulation. Here, a tunable biomimetic hydrogel system that possesses alterable gelation kinetics and biologically relevant matrix stiffness is developed to study 3D dynamic cellular responses during encapsulation. Hydrogels are synthesized by cross-linking thiolated hyaluronic acid and thiolated chondroitin sulfate with polyethylene glycol diacrylate under cell-compatible conditions. Hydrogel properties are tailored by altering thiol substitution degrees of glycosaminoglycans or molecular weights of cross-linkers. Encapsulation of human mesenchymal stem cells through hydrogel gelation reveals high cell viability as well as a three-stage gelation-dependent cellular response in real-time focal adhesion kinase (FAK) phosphorylation in live single cells. Furthermore, stiffer hydrogels result in higher equilibrium FAK activity and enhanced actin protrusions. Our results demonstrate the promise of hydrogel-mediated cellular responses during cell encapsulation.


Genetic analysis of the Caenorhabditis elegans pax-6 locus: roles of paired domain-containing and nonpaired domain-containing isoforms.
PAX-6 proteins are involved in eye and brain development in many animals. In the nematode Caenorhabditis elegans the pax-6 locus encodes multiple PAX-6 isoforms both with and without a paired domain. Mutations in the C. elegans pax-6 locus can be grouped into three classes. Mutations that affect paired domain-containing isoforms cause defects in epidermal morphogenesis, epidermal cell fates, and gonad cell migration and define the class I (vab-3) complementation group. The class II mutation mab-18(bx23) affects nonpaired domain-containing isoforms and transforms the fate of a sensory organ in the male tail. Class III mutations affect both paired domain and nonpaired domain isoforms; the most severe class III mutations are candidate null mutations in pax-6. Class III mutant phenotypes do not resemble a simple sum of class I and class II phenotypes. A comparison of class I and class III phenotypes indicates that PAX-6 isoforms can interact additively, synergistically, or antagonistically, depending on the cellular context.


In-field assessment of sodium oxybate effect in pediatric type 1 narcolepsy: an actigraphic study.
Sodium oxybate (SXB) is a GABAergic agent widely used as off-label treatment in pediatric type 1 narcolepsy (NT1). Here, we aimed at analyzing by wrist actigraphy the sleep/wake profile of NT1 children and adolescents in drug-naïve condition and after 1 year of SXB treatment. As secondary aim, we investigated changes on sleepiness, cataplexy, and children's anthropometric profile after 1 year of SXB treatment. Twenty-four drug-naïve NT1 children underwent 7 days of actigraphy during the school week. Information on sleepiness, narcolepsy symptoms, and anthropometric features were collected during the same week with questionnaires and semistructured clinical interview. Children started SXB treatment and underwent a second evaluation encompassing actigraphy, clinical interview, questionnaires, and anthropometric assessment after 1 year of stable treatment. Actigraphy effectively documented an improvement of nocturnal sleep quality and duration coupled with a reduction of diurnal total sleep time, nap frequency, and duration at 1 year follow-up. Reduction of sleepiness, cataplexy frequency and severity, and weight loss, mainly in obese and overweight NT1 children, were also observed at the 1 year follow-up. Actigraphy objectively documented changes in nocturnal sleep quality and diurnal napping behavior after 1 year of SXB treatment, thus representing a valid approach to ecologically assess SXB treatment effect on NT1 children's sleep/wake profile. NT1 symptoms severity and children's anthropometric features also changed as expected. Actigraphy offers the possibility to longitudinally follow up children and has potential to become a key tool to tailor treatment in pediatric patients.


Epiplexus cells in the embryos of the turtle, Trionyx sinensis.
The epiplexus cells in the embryos of the turtle, Trionyx sinensis were examined in scanning and transmission electron microscopy. Although the cells showed diverse morphological forms, the majority of the cells however had a stellate cell body each with 3-5 radiating long processes with secondary processes terminating on the ventricular surface of the choroid epithelial cells. The cell body was relatively smooth. Occasional epiplexus cells displayed blebs on their cell bodies and processes. In the transmission electron microscope, the nucleus contained coarse chromatin clumps. The cytoplasm was endowed with large lysosomal bodies. Based on these observations, it is suggested the epiplexus cells in the turtle embryos are phagocytic even though most of them probably are at the resting stage.


Control of response selection by reinforcer value requires interaction of amygdala and orbital prefrontal cortex.
Goal-directed actions are guided by expected outcomes of those actions. Humans with bilateral damage to ventromedial prefrontal cortex, or the amygdala, are deficient in their ability to use information about positive and negative outcomes to guide their choice behavior. Similarly, rats and monkeys with orbital prefrontal or amygdala damage have been found to be impaired in their responses to changing values of outcomes. In the present study, we tested whether direct, functional interaction between the amygdala and the orbital prefrontal cortex is necessary for guiding behavior based on expected outcomes. Unlike control monkeys, rhesus monkeys with surgical disconnection of these two structures, achieved by crossed unilateral lesions of the amygdala in one hemisphere and orbital prefrontal cortex in the other, combined with forebrain commissurotomy, were unable to adjust their choice behavior after a change in the outcome (here, a reduction in the value of a particular reinforcer). The lesions did not affect motivation to work for a food reinforcer, or food preferences, per se. Hence, the amygdala and orbital prefrontal cortex act as part of an integrated neural system guiding decision-making and adaptive response selection.


Determination of death in donation after circulatory death: an ethical propriety.
The recently developed donation after circulatory death (DCD) heart transplant technique, pioneered by Papworth Hospital in the UK, involves the use of extracorporeal perfusion technologies to restart the donor heart in situ and thereby restore the donor's own circulation, after first isolating the donor's cerebral circulation. By restoring the circulation in the deceased donor, even if the cerebral circulation is excluded, the Papworth technique challenges the acceptability of death determination in DCD. This study uses as its exemplar case the Papworth DCD heart technique to review and make wider comment about death determination in DCD. We seek to answer three challenges to ethical propriety raised by the Papworth technique: death determination using the permanence standard (common to all DCD practice); restoration of heart contractility and circulation in the body; and active prevention of the restoration of brain circulation by use of a cross-clamp to isolate the cerebral circulation. The Papworth technique for heart DCD does not compromise the permanence standard for declaring death and therefore respects the dead donor rule in the UK, but perhaps elsewhere the law would need to change to refer to the cessation of circulation in the brain.


Multispecies Probiotic Supplementation Favorably Affects Vascular Function and Reduces Arterial Stiffness in Obese Postmenopausal Women-A 12-Week Placebo-Controlled and Randomized Clinical Study.
Obesity in the postmenopausal period is associated with an increased risk of cardiovascular diseases in women. One of the key drivers of cardiovascular risk is endothelial dysfunction; thus, this is also a crucial point for studies on new therapeutic methods of cardioprotective properties. The aim of the current study was to evaluate the effect of two doses of multispecies probiotic Ecologic® Barrier supplement on functional (primary endpoint) and biochemical parameters (secondary endpoint) of endothelial dysfunction in obese postmenopausal women in a 12-week randomized, placebo-controlled clinical trial. A total of 81 obese Caucasian women participated in the trial. The subjects were randomly assigned to three groups that received a placebo, a low dose (LD) (2.5 × 10⁸ colony forming units (CFU) per day), or a high dose (HD) (1 × 1010 CFU per day) of lyophilisate powder containing live multispecies probiotic bacteria. The probiotic supplement was administered each day for 12 weeks in two equal portions. A high dose probiotic supplementation for 12 weeks decreased systolic blood pressure, vascular endothelial growth factor, pulse wave analysis systolic pressure, pulse wave analysis pulse pressure, pulse wave analysis augmentation index, pulse wave velocity, interleukin-6, tumor necrosis factor alpha, and thrombomodulin. Low doses of probiotic supplementation decreased the systolic blood pressure and interleukin-6 levels. The mean changes in the estimated parameters, compared among the three groups, revealed significant differences in the vascular endothelial growth factor, the pulse wave analysis systolic pressure, the pulse wave analysis augmentation index, the pulse wave velocity, the tumor necrosis factor alpha, and thrombomodulin. The post hoc tests showed significant differences for all parameters between HD and the placebo group, and HD and LD (besides pulse wave analysis augmentation index). We show for the first time that supplementation with multispecies probiotic Ecologic® Barrier favorably modifies both functional and biochemical markers of vascular dysfunction in obese postmenopausal women.


Devices used by automated milking systems are similarly accurate in estimating milk yield and in collecting a representative milk sample compared with devices used by farms with conventional milk recording.
Information on accuracy of milk-sampling devices used on farms with automated milking systems (AMS) is essential for development of milk recording protocols. The hypotheses of this study were (1) devices used by AMS units are similarly accurate in estimating milk yield and in collecting representative milk samples compared with devices used by certified milk recording providers on farms with conventional milking systems (CMS) and (2) devices used on both AMS and CMS comply with accuracy criteria described by the New Zealand Standard and by the International Committee of Animal Recording (ICAR). Milk recording data from 5 AMS farms were collected during 13 milk recording test days between December 2011 and February 2013. Milk yield was estimated by ICAR-approved milk meters on AMS units. Milk samples were collected over a 48-h period and submitted to an off-site certified laboratory for milk composition analysis. Data were also collected manually from 5 to 10 cows per AMS unit; a complete milking of a cow was weighed to serve as gold standard for milk yield, and 3 milk samples per cow milking were collected and analyzed in the laboratory to serve as gold standards for milk composition. A similar procedure was used during 6 milk recording occasions with devices used during conventional milk recording at a CMS research farm. Farm type, breed, season, and region did not appear to affect accuracy of devices used on AMS units. Milk meters used by AMS units complied with ICAR limits in 12.5 and 25% of the milk recording test days for test bucket weights between 2 and 10kg and for test bucket weights >10kg, respectively. These percentages were 52 and 42%, respectively, for devices used on CMS. Analyzing all samples as one milk recording test day, 1.4% fell outside the 20% difference band for AMS compared with 1.1% of the milk samples for CMS. Devices used by AMS complied with ICAR in 73% of the milk recording test days for fat percentage, compared with 42% of the milk recording test days by devices used at the CMS farm. When analyzing all milk samples as one milk recording test day, 3.5% of the milk samples fell outside the 99% ICAR limit for AMS compared with 17.2% of the milk samples for CMS. Applying the ICAR standards for fat percentage to crude protein percentage and SCC, devices used on AMS were accurate in estimating crude protein percentage but not in estimating SCC. Thus, devices on AMS units did not comply with national nor ICAR standards with regard to milk yield and fat percentage. However, devices used on AMS were similarly or more accurate compared with devices used during conventional milk recording. It is proposed that devices used on AMS units, when calibrated regularly and when set up according to the manufacturer's instruction, have similar or improved accuracy compared with CMS devices. Because the New Zealand industry accepts data from devices currently used by certified providers for milk recording on CMS farms, results imply that the AMS devices should also be permitted to be used for milk recording.


Multiple descriptions coinciding lattice vector quantizer for wavelet image coding.
Multiple description (MD) coding has been a popular choice for robust data transmission over the unreliable network channels. Lattice vector quantization provides lower computation for efficient data compression. In this paper, a new MD coinciding lattice vector quantizer (MDCLVQ) is presented. The design of the quantizer is based on coinciding 2-D hexagonal sublattices. The coinciding sublattices are geometrically similar sublattices, with the same index but generated by different generator matrices. A novel labeling algorithm based on the hexagonal coinciding sublattices is also developed. Performance results of the MDCLVQ scheme, together with the new labeling algorithm applied to standard test images, show improvements of the central and side decoders, as compared with the renowned techniques for several test images.


Vitamin requirements of intravenously fed man.
The continued refinements in the application of intravenous feeding must include the provision of optimal micronutrients if the clinical utility of this technique is to be fully realized. From the above discussion, it is readily apparent that our understanding of vitamin requirements during IVF is fragmentary at best. This limited knowledge has unfortunately been extrapolated to the daily regimen of nearly all parenterally nourished patients. The fact that this is done without apparent ill effect should not deter future investigation, particularly in the critically injured or organ failure patient where subtle vitamin deficiencies may be masked by concurrent illnesses. Research should be directed toward the assessment of tissue vitamin concentration and function, with correlations to the degree of energy expenditure and/or nitrogen turnover exhibited by the patient.


Acute anterior myocardial infarction in a young male patient homozygous for the factor V Leiden mutation.
There are several reports on the association between the factor V Leiden mutation and acute myocardial infarction (AMI) in young patients, in particular young males. A 28-year-old male patient was admitted with severe chest pain of new onset. He was an active smoker. His father had a history of coronary artery disease and AMI after the age of 45 years. There were no other major coronary risk factors. His electrocardiogram showed ST-segment elevation in the precordial leads V1 to V5. His blood pressure, pulse rate, and other clinical parameters were stable. Emergency coronary angiography showed a significant narrowing in the mid-portion of the left anterior descending (LAD) artery with a moderate intracoronary thrombus, and no or minimal atherosclerosis. The other coronary arteries were normal. Direct stenting was performed for the culprit lesion, which resulted in relief of obstruction and significant improvement in the LAD artery. DNA samples isolated from the peripheral blood were analyzed by polymerase chain reaction and the patient was found to be homozygous for the factor V Leiden mutation. Transthoracic echocardiography before discharge showed only mild hypokinesis of the anterior and apical segments.


Interaction of atropine with naloxone to alter anterior pituitary hormone secretion.
The effects of third ventricular injection of naloxone and atropine, alone and together, on plasma concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), growth hormone (GH), and thyroid-stimulating hormone (TSH) were studied in conscious, ovariectomized (OVX) rats. Samples were removed from unrestrained rats that had been previously implanted with an atrial cannula, and plasma hormone levels were determined by radioimmunoassay. Intraventricular injection of naloxone (30 micrograms) induced increases in plasma LH and GH concentrations, decreased plasma PRL levels, but did not alter plasma TSH. When atropine (100 micrograms) was injected intraventricularly, there was a decrease in plasma LH and an increase in PRL levels but no alterations in the levels of the other hormones. The action of naloxone on PRL and GH release was eliminated by simultaneous administration of atropine, but there was only a slight reduction in its effect on LH. The results indicate that central cholinergic neurons may play an important role, via muscarinic type receptors, in mediation of the effects of naloxone and presumably opiates on PRL and GH release, whereas the effects on LH appear to proceed independently of cholinergic neurons.


[Activities in daily life and changes in care level among users of Preventive Care Service under Long-Term Care Insurance].
The aim of this study was to investigate the association between social roles (engaged in working outside or house work) or hobbies and changes in care level among users of Preventive Care Service under Long-Term Care Insurance. A total of 8,734 Preventive Care Service users were analyzed. The assessment was conducted between February 2007 and December 2008. The improvement, maintenance, and aggravation in care level were defined by changes in care level from the beginning to the end. To assess their activities in daily life, the participants were asked, "Which is the major activity in your daily life? Please select one from the following: working outside, house work, hobbies, watching television, others, or none." We used the multiple logistic regression model to calculate the odds ratio (OR) and 95% confidence interval (CI) of the maintenance or improvement in care level according to the category of activity. As compared with subjects who answered none, the multivariate adjusted ORs (95% CI) of the maintenance or improvement in care level were 2.0 (1.4-2.9) for those who answered working outside, 1.5 (1.2-1.8) for those who answered house work, and 1.5 (1.2-1.9) for those who answered hobbies. However, those who answered watching television or others did not show a significant association with the maintenance or improvement in care level. In this study, compared with subjects who answered none, those who answered that they were engaged in working outside, house work, or hobbies were associated with having significantly higher ORs of the maintenance or improvement in care level.


Alcohol dehydrogenases from Kluyveromyces marxianus: heterologous expression in Escherichia coli and biochemical characterization.
Kluyveromyces marxianus has recently become a species of interest for ethanol production since it can produce ethanol at high temperature and on a wide variety of substrates. However, the reason why this yeast can produce ethanol at high temperature is largely unknown. The ethanol fermentation capability of K. marxianus GX-UN120 at 40°С was found to be the same as that of Saccharomyces cerevisiae at 34°С. Zymogram analysis showed that alcohol dehydrogenase 1 (KmAdh1) was largely induced during ethanol production, KmAdh4 was constitutively expressed at a lower level and KmAdh2 and KmAdh3 were almost undetectable. The genes encoding the four alcohol dehydrogenases (ADHs) were cloned from strain GX-UN120. Each KmADH was expressed in Escherichia coli and each recombinant protein was digested with enterokinase to remove the fusion protein. The optimum pH of the purified recombinant KmAdh1 was 8.0 and that of KmAdh2, KmAdh3 and KmAdh4 was 7.0. The optimum temperatures of KmAdh1, KmAdh2, KmAdh3 and KmAdh4 were 50, 45, 55 and 45°C, respectively. The K(m) values of the recombinant KmAdh1 and KmAdh2 were 4.0 and 1.2 mM for acetaldehyde and 39.7 and 49.5 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh1 and KmAdh2 were 114.9 and 21.6 μmol min⁻¹ mg⁻¹ for acetaldehyde and 57.5 and 1.8 μmol min⁻¹ mg⁻¹ for ethanol, respectively. KmAdh3 and KmAdh4 catalyze the oxidation reaction of ethanol to acetaldehyde but not the reduction reaction of acetaldehyde to ethanol, and the K(m) values of the recombinant KmAdh3 and KmAdh4 were 26.0 and 17.0 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh3 and KmAdh4 were 12.8 and 56.2 μmol min⁻¹ mg⁻¹ for ethanol, respectively. These data in this study collectively indicate that KmAdh1 is the primary ADH responsible for the production of ethanol from the reduction of acetaldehyde in K. marxianus. The relatively high optimum temperature of KmAdh1 may partially explain the ability of K. marxianus to produce ethanol at high temperature. Understanding the biochemical characteristics of KmAdhs will enhance our fundamental knowledge of the metabolism of ethanol fermentation in K. marxianus.


Sexual behavior associated with low Verbal IQ in youth who have severe mental illness.
A retrospective chart review was used to examine sexual behavior (hypersexual, exposing, and victimizing) and cognitive impairment in 200 youth who had serious mental illness. Lower IQ was associated with increased sexual acting-out. For more serious victimizing sexual behaviors, only Verbal IQ differences reached statistical significance. Overall, sexual behavior was strongly associated with a history of sexual abuse. Sexual abuse history was significantly associated only with lower Performance IQ. Therefore, the association between low Verbal IQ and sexual victimizing behavior is distinct from the effects of sexual abuse. Results underscore the importance of verbal cognitive abilities, regardless of overall cognitive level, in the etiology and treatment of sexual behaviors, especially among individuals without a history of sexual abuse.


Localization of human coagulation factor VIII (hFVIII) in transgenic rabbit by FISH-TSA: identification of transgene copy number and transmission to the next generation.
For chromosomal localization of the hFVIII human transgene in F2 and F3 generation of transgenic rabbits, FISH-TSA was applied. A short cDNA probe (1250 bp) targeted chromosomes 3, 7, 8, 9 and 18 of an F2 male (animal 1-3-8). Two transgenic offspring (F3) revealed signal positions in chromosome 3 and chromosomes 3 and 7, respectively. Sequencing and structure analysis of the rabbit orthologous gene revealed high similarity to its human counterpart. Part of the sequenced cDNA (1310 bp) served as a probe for FISH-TSA analysis. The rabbit gene was localized in the q arm terminus of the X chromosome. This result is in agreement with reciprocal chromosome painting between the rabbit and the human. The presented FISH-TSA method provides strong signals without any interspecies reactivity.