Synapsin I Controls Synaptic Maturation of Long-Range Projections in the Lateral Amygdala in a Targeted Selective Fashion.
The amygdala, and more precisely its lateral nucleus, is thought to attribute emotional valence to external stimuli by generating long-term plasticity changes at long-range projections to principal cells. Aversive experience has also been shown to modify pre- and post-synaptic markers in the amygdala, suggesting their possible role in the structural organization of adult amygdala networks. Here, we focused on how the maturation of cortical and thalamic long-range projections occurs on principal neurons and interneurons in the lateral amygdala (LA). We performed dual electrophysiological recordings of identified cells in juvenile and adult GAD67-GFP mice after independent stimulation of cortical and thalamic afferent systems. The results demonstrate that synaptic strengthening occurs during development at synapses projecting to LA principal neurons, but not interneurons. As synaptic strengthening underlies fear conditioning which depends, in turn, on presence and increasing expression of synapsin I, we tested if synapsin I contributes to synaptic strengthening during development. Interestingly, the physiological synaptic strengthening of cortical and thalamic synapses projecting to LA principal neurons was virtually abolished in synapsin I knockout mice, but not differences were observed in the excitatory projections to interneurons. Immunohistochemistry analysis showed that the presence of synapsin I is restricted to excitatory contacts projecting to principal neurons in LA of adult mice. These results indicate that synapsin I is a key regulator of the maturation of synaptic connectivity in this brain region and that is expression is dependent on postsynaptic identity.


Evaluation of Selected Markers in Kidneys of Cynomolgus Monkey (Macaca fascicularis) with Induced Diabetes during Renal Ischemia-reperfusion Injury.
We previously reported that induced type 1 diabetes mellitus (DM) increases the susceptibility of acute kidney injury in- duced by ischemia-reperfusion injury (IRI) in cynomolgus monkeys. In this follow-up study, we compared the expression of selected markers in the renal tissues of monkeys subjected to bilateral renal IRI with and without diabetes. All tissues were obtained from the original study. Renal biopsies were obtained before and 24 and 48 h after ischemia and were examined for expression of KI-67 (tubular proliferation), Na+ /K+ ATPase (sodium-potassium pump), TNF-α(tumor necrosis factor-α, inflammation), CD31 (microvessels), CD3 (T-cells), 2 fibrotic markers (fibroblast specific protein-1, FSP-1;α-smooth muscle actin,α -SMA), and cleaved caspase 3 (apoptosis). Generally, the expression of these markers differed in monkeys with and without DM. As compared with non-DM monkeys, DM monkeys had more cells that expressed KI-67 during progression of acute kidney injury (AKI). Na+ /K+ ATPase expression was clearly present at baseline in the basolateral tubular areas only in the non-DM monkeys. At 48 h, its expression in the basolateral area was not visible in DM monkeys, but was still present in intercellular junctions of non-DM monkeys. The expression of TNF-αwas higher in DM before and 48 h after ischemia. Before and 24 h after ischemia, the number of CD31-positive capillaries was not different between 2 groups, although more collapsed vessels were found at in DM at 24 h. At 48 h, the number of capillaries was less in DM compared with those from non-DM animals. DM monkeys had more interstitial CD3-positive cells than did non-DM monkeys at 24 and 48 h after ischemia. Finally, FSP-1-stained cells were more abundant in DM than non-DM at 24 and 48 h. Our results show that DM aggravates the recovery of renal ischemia/reperfusion injury by affecting tubular proliferation, capillary density, T cell infil- tration and by altering protein and mRNA expression of various genes involved in ion channel, inflammation, and fibrotic change. The results from this observational study demonstrate that DM aggravates the recovery of renal ischemia/reperfusion injury by affecting multiple events including tubular necrosis, proliferation, function, inflammation and by inducing capillary rarefaction in cynomolgus monkeys.


Chemoresistance in high-grade gliomas: relevance of adenosine signalling in stem-like cells of glioblastoma multiforme.
Glioblastoma multiforme (GBM) is the most common glial cell-derived brain tumour, with one of the worst prognoses among all cancers. GBM cells are infiltrative and extremely resistant to radio- and chemotherapy, which inevitably leads to recurrence after surgical resection. These inherent GBM properties are the reasons that patient treatment has not seen major improvements in decades. Studies have consistently shown that glioblastoma stem-like cells (GSCs) are responsible for the tumourigenic properties in the GBM population. In fact, their self-renewal and proliferative potential are required for tumour growth, and their extreme chemoresistance leads to early recurrence of this tumour. Among those mechanisms associated with chemoresistance and having the greatest clinical impact in cancer treatment, are the activities of plasma membrane transporters that extrude antitumour drugs from the cell, thus notably decreasing the pharmacological efficiency of these drugs. The multiple drug resistance associated protein-1 (Mrp1) transporter has been shown to be particularly important in GBM, as inhibition of Mrp1 activity notably chemosensitises cells to antiproliferative drugs. As current therapeutic options for GBM offer only a poor improvement in overall survival rate, alternative strategies for overcoming tumour resistance are urgently sought after. To this end, it is of major clinical relevance to know more about the endogenous modulators that control Mrp1 expression within the pathological environment of the tumour. This review describes the particular properties of glioblastoma cells that overcome multimodal therapy and relapse, with an emphasis on the microenvironmental tumour properties that influence the chemoresistance phenotype to antiproliferative drugs. We also discuss alternative methods of reversal of Mrp1-mediated chemoresistance in these cells by targeting extracellular adenosine production or signalling through particular plasma membrane receptors.


Prediction of Intrinsic Disorder in MERS-CoV/HCoV-EMC Supports a High Oral-Fecal Transmission.
A novel coronavirus, MERS-CoV (NCoV, HCoV-EMC/2012), originating from the Middle-East, has been discovered. Incoming data reveal that the virus is highly virulent to humans. A model that categorizes coronaviuses according to the hardness of their shells was developed before the discovery of MERS-CoV. Using protein intrinsic disorder prediction, coronaviruses were categorized into three groups that can be linked to the levels of oral-fecal and respiratory transmission regardless of genetic proximity. Using this model, MERS-CoV is placed into disorder group C, which consists of coronaviruses that have relatively hard inner and outer shells. The members of this group are likely to persist in the environment for a longer period of time and possess the highest oral-fecal components but relatively low respiratory transmission components. Oral-urine and saliva transmission are also highly possible since both require harder protective shells. Results show that disorder prediction can be used as a tool that suggests clues to look for in further epidemiological investigations.


Do hair bulb melanocytes undergo apoptosis during hair follicle regression (catagen)?
The fate of the hair follicle pigmentary unit during the cyclical involution of anagen hair follicles is unknown. Using the C57BL/6 mouse model for hair research, hair follicle melanocytes were examined during the anagen-catagen transformation, comparing spontaneous and pharmacologically induced catagen development. This study shows that both spontaneous catagen and dexamethasone-induced catagen display similar changes in the pigmentary unit. Catagen hair follicles exhibited pigment incontinence in the dermal papilla and in selected outer root sheath keratinocytes. Melanocytes deleted by apoptosis were detected in spontaneous catagen and, more commonly, in dexamethasone-induced catagen, and were identified using transmission electron microscopy by the presence of free premelanosomes in affected cells lacking epithelial specializations, and by the colocalization of TUNEL positivity and tyrosinase-related protein-1 immunoreactivity. By contrast, cyclophosphamide-induced catagen was characterized by the initial retention of melanogenic and dendritic melanocytes in the presence of widespread keratinocyte apoptosis. Melanocyte incontinence and the ectopic distribution of melanin were more severe than in the other forms of catagen. Whereas much of this melanin was extruded, via the hair canal, to the skin surface, hair follicle-derived pigment was also detected within the epidermis, probably derived from pigment-carrying migrating outer root sheath keratinocytes from the proximal hair follicle. Thus, apoptosis may account, at least in part, for the loss of melanogenic melanocytes during spontaneous catagen. Although dexamethasone-induced catagen may provide a useful model for general hair pigmentation research, catagen induced by cyclophosphamide offers an interesting model for studying the response, and relative resistance, of melanocytes to chemical injury.


Noninvasive indices of fibrosis in NAFLD: starting to think about a three-hit (at least) phenomenon.
NAFLD is a highly prevalent, phenotypically diverse condition. A small minority of patients with NAFLD progress to clinically important stages of fibrosis. Identifying patients at risk for progressive disease is one of the great challenges of hepatology. There are an increasing number of models that have been reported to provide noninvasive indices of histological severity of NAFLD, including a new report by Gholam et al. in this issue of the Journal. All models estimating risk of severity of NAFLD lack predictivity for future progression of fibrosis. Although conceptually considered to be a "two-hit" phenomenon, the combination of steatosis, necroinflammation, and progressive fibrosis is likely to be much more complex. Because the phenotypic expression of NAFLD, like so many liver diseases, is an interaction among genes, behavior, and environment, predicting future outcomes for patients with NAFLD (and liver diseases in general) will likely require models that combine genetic susceptibility, biological environment, and behavior. Emerging data are reviewed that suggest that it will not be long before studies like that of Gholam et al. are combined with those of clinical genomics to deliver tools that we can apply to the individual patient.


Cord blood brain derived neurotrophic factor: diagnostic and prognostic marker in fullterm newborns with perinatal asphyxia.
This prospective case control study was designed to evaluate cord blood brain derived neurotrophic factor level in full term newborns with perinatal asphyxia as a marker of central nervous system insult and predictor of severity of hypoxic ischemic encephalopathy, with follow up of its level during the reperfusion phase. The study included twenty fullterm neonates with perinatal asphyxia (cases) and twenty controls. Cord blood samples were obtained at birth and peripheral blood samples at 72 h postnatal from cases only. Plasma brain derived neurotrophic factor level was measured using enzyme linked immunosorbent assay. The clinical severity of encephalopathy was graded based on Sarnat and Sarnat staging. Cord Plasma brain derived neurotrophic factor level was significantly increased among cases compared to controls. Among cases, brain derived neurotrophic factor level at delivery and after 72 h significantly correlated with the severity of encephalopathy according to Sarnat staging being higher as severity increases. Brain derived neurotrophic factor level significantly increased after 72 h of life compared to its level at delivery among cases. Brain derived neurotrophic factor levels at delivery and at 72 h postnatal were predictors of severe Sarnat stage and poor outcome. We concluded that brain derived neurotrophic factor level as a marker of central nervous system insult is increased in full term newborns with perinatal asphyxia. It can serve as an indicator for the severity of encephalopathy and adverse outcomes.


Two-dimensional metamaterial structure exhibiting reduced visibility at 500 nm.
Metamaterials provide unprecedented freedom and flexibility in the creation of new structures, which control electromagnetic wave propagation in very unusual ways. Very recently various theoretical designs for an electromagnetic cloak were suggested and an experimental realization of a partial cloak operating in a two-dimensional cylindrical geometry were reported in the microwave frequency range. We report on an experimental two-dimensional reduced visibility structure that approximates the distribution of the radial component of the dielectric permittivity necessary to achieve nonmagnetic cloaking in the visible frequency range.


Automatic image orientation detection.
We present an algorithm for automatic image orientation estimation using a Bayesian learning framework. We demonstrate that a small codebook (the optimal size of codebook is selected using a modified MDL criterion) extracted from a learning vector quantizer (LVQ) can be used to estimate the class-conditional densities of the observed features needed for the Bayesian methodology. We further show how principal component analysis (PCA) and linear discriminant analysis (LDA) can be used as a feature extraction mechanism to remove redundancies in the high-dimensional feature vectors used for classification. The proposed method is compared with four different commonly used classifiers, namely k-nearest neighbor, support vector machine (SVM), a mixture of Gaussians, and hierarchical discriminating regression (HDR) tree. Experiments on a database of 16 344 images have shown that our proposed algorithm achieves an accuracy of approximately 98% on the training set and over 97% on an independent test set. A slight improvement in classification accuracy is achieved by employing classifier combination techniques.


Decreasing sperm quality in mice subjected to chronic cannabidiol exposure: New insights of cannabidiol-mediated male reproductive toxicity.
Cannabidiol (CBD) is a natural cannabinoid present in the Cannabis sativa plant, widely prescribed as an anticonvulsant drug, especially for pediatric use. However, its effects on male reproduction are still little investigated. Therefore, the present study assessed the effects of CBD on the spermatogenesis and sperm quality. For this, twenty-one-day-old Swiss mice received CBD for 34 consecutive days by gavage at doses of either 15 or 30 mg/kg. Chronic exposure to CBD decreased the frequency of stages VII-VIII and XII of spermatogenesis and an increase in the frequency of stage IX were noted. Furthermore, the seminiferous epithelium height reduced at stage IX and increased at stage XII in both CBD-treated groups. There was a significant rise of sperm DNA damage, while no genotoxic effects were observed in leukocytes. The activities of superoxide dismutase and catalase decreased, while malondialdehyde levels increased in the sperm of mice treated with a higher dose of CBD. Mice exposed to 30 mg/kg of CBD showed a reduction in the mobile spermatozoa percentage and in curvilinear velocity, while straight line and average path velocity decreased in both treated groups. The number of acrosome-intact spermatozoa declined in the CBD 30 group, and the number of abnormal acrosomes raised in both CBD groups. On the other hand, the weight of reproductive organs, sperm count, and hormone levels were not affected by CBD treatment. These findings show that dysregulation of the endocannabinoid system by CBD can reduce sperm quality. The mechanisms responsible may be associated with disorders during spermatogenesis, especially during the final stages of nuclear remodelling and assembly of acrosome. However, changes in mitochondrial function, as well as the reduction on the antioxidant enzyme activities during epididymal transit, at least partly, may also be involved.


Ulnar artery thrombosis associated with anomalous hypothenar muscle.
Ulnar artery thrombosis should be considered in patients presenting with cold intolerance or ischemia of the small and ring fingers, or a mass in the hypothenar area. Frequently this diagnosis is associated with a history of repetitive blunt trauma to the ulnar hand, thereby traumatizing the ulnar artery in Guyon's canal. This report presents a case of ulnar artery thrombosis associated with an abnormal muscle originating on the palmar antebrachial fascia, traversing volar to the ulnar artery and nerve, and inserting on the ulnar border of the abductor digiti minimi. This muscle is an abductor digiti minimi accessorius.


A method of recognition of arabic cursive handwriting.
In spite of the progress of machine recognition techniques of Latin, Kana, and Chinese characters over the two past decades, the machine recognition of Arabic characters has remained almost untouched. In this correspondence, a structural recognition method of Arabic cursively handwritten words is proposed. In this method, words are first segmented into strokes. Those strokes are then classified using their geometrical and topological properties. Finally, the relative position of the classified strokes are examined, and the strokes are combined in several steps into a string of characters that represents the recognized word. Experimental results on texts handwritten by two persons showed high recognition accuracy.


Unemployment Insurance and Disability Insurance in the Great Recession.
Social Security Disability Insurance (SSDI) awards rise during recessions. If marginal applicants are able to work but unable to find jobs, countercyclical Unemployment Insurance (UI) benefit extensions may reduce SSDI uptake. Exploiting UI extensions in the Great Recession as a source of variation, we find no indication that expiration of UI benefits causes SSDI applications and can rule out effects of meaningful magnitude. A supplementary analysis finds little overlap between the two programs' recipient populations: only 28% of SSDI awardees had any labor force attachment in the prior calendar year, and of those, only 4% received UI.


Higher Plasma Selenium Concentrations Are Associated with Increased Odds of Prevalent Type 2 Diabetes.
Selenium, an essential trace element, has been investigated as a potential cancer prevention agent. However, several studies have indicated that selenium supplementation may be associated with an increased risk of type 2 diabetes (T2D), although an equivocal relation of this nature requires confirmation. We examined the association between baseline plasma concentrations of selenium and the prevalence of T2D, as well as whether participant characteristics or intake of other antioxidant nutrients modified this relation. We conducted cross-sectional analyses of 1727 participants from the Selenium Trial, a randomized clinical trial of selenium supplementation for colorectal adenoma chemoprevention that had data for baseline selenium plasma concentrations, T2D status, and dietary intake. Logistic regression modeling was used to evaluate the associations between plasma selenium concentrations and prevalent T2D, adjusting for confounding factors. Heterogeneity of effect by participant characteristics was evaluated utilizing likelihood-ratio tests. Mean ± SD plasma selenium concentrations for those with T2D compared with those without T2D were 143.6 ± 28.9 and 138.7 ± 27.2 ng/mL, respectively. After adjustment for confounding, higher plasma selenium concentrations were associated with a higher prevalence of T2D, with ORs (95% CIs) of 1.25 (0.80, 1.95) and 1.77 (1.16, 2.71) for the second and third tertiles of plasma selenium, respectively, compared with the lowest tertile (P-trend = 0.007). No significant effect modification was observed for age, sex, body mass index, smoking, or ethnicity. Increased odds of T2D were seen among those who were in the highest tertile of plasma selenium and the highest category of intake of β-cryptoxanthin (P-trend = 0.03) and lycopene (P-trend = 0.008); however, interaction terms were not significant. These findings show that higher plasma concentrations of selenium were significantly associated with prevalent T2D among participants in a selenium supplementation trial. Future work is needed to elucidate whether there are individual characteristics, such as blood concentrations of other antioxidants, which may influence this relation.


Usefulness of admission gamma-glutamyltransferase level for predicting new-onset heart failure in patients with acute coronary syndrome with left ventricular systolic dysfunction.
Our aim was to determine whether there is a relationship between admission gamma-glutamyltransferase (GGT) and subsequent heart failure hospitalizations in patients with acute coronary syndrome. We selected 123 patients with newly diagnosed acute coronary syndrome of ejection fraction (EF) <45%. Patients were followed 15±10 months, and the relationship between admission GGT level and hospitalization because of heart failure during the follow-up was examined. Twenty-three (18.7%) patients were hospitalized during the follow-up of 15±10 months. Receiver operating characteristic (ROC) curve analysis showed that the cut-off point of admission GGT related to predict hospitalization was 49 IU/L, with a sensitivity of 81.7% and specificity of 65.2%. Increased GGT >49 IU/L on admission, presence of hypertension and hyperlipidemia, left ventricular ejection fraction (LVEF), right ventricular dysfunction, moderate-to-severe mitral regurgitation, alanine aminotransferase level, and antiplatelet agent usage were found to have prognostic significance in univariate Cox proportional hazards analysis. In multivariate Cox proportional-hazards model, increased GGT >49 IU/L on admission (hazard ratio [HR] 2.663, p=0.047), presence of hypertension (HR 4.107, p=0.007), and LVEF (HR 0.911, p=0.002) were found to be independent factors to predict new-onset heart failure requiring hospitalization. Hospitalization in heart failure was associated with increased admission GGT levels. Increased admission GGT level in acute coronary syndrome with heart failure should be monitored closely and treated aggressively.


New developments in optimizing bronchodilator treatment of COPD: a focus on glycopyrrolate/formoterol combination formulated by co-suspension delivery technology.
COPD causes considerable health and economic burden worldwide, with incidence of the disease expected to continue to rise. Inhaled bronchodilators, such as long-acting muscarinic antagonists (LAMAs) and long-acting β2-agonists (LABAs), are central to the maintenance treatment of patients with COPD. Clinical studies have demonstrated that combined LAMA + LABA therapies improve efficacy while retaining a safety profile similar to LAMA or LABA alone. This has led to the development of several LAMA/LABA fixed-dose combination (FDC) therapies, which provide patients with the convenience of two active compounds in a single inhaler. GFF MDI (Bevespi Aerosphere®) is an FDC of glycopyrrolate/formoterol fumarate 18/9.6 µg formulated using innovative co-suspension delivery technology for administration via metered dose inhaler (MDI). GFF MDI was developed to make a treatment option available for patients who have a requirement or preference to use an MDI, rather than a dry powder or soft mist inhaler. Now that several LAMA/LABA FDCs have been approved for use in COPD, we review the impact of dual-bronchodilator treatment on COPD therapy and discuss recent clinical studies that are helping to develop a more comprehensive understanding of how LAMA/LABA FDCs can improve patient outcomes.


Glycine max Fermented by a Novel Probiotic, Bifidobacterium animalis subsp. lactis LDTM 8102, Increases Immuno-Modulatory Function.
Many probiotic species have been used as a fermentation starter for manufacturing functional food materials. We have isolated Bifidobacterium animalis subsp. lactis LDTM 8102 from the feces of infants as a novel strain for fermentation. While Glycine max has been known to display various bioactivities including anti-oxidant, anti-skin aging, and anti-cancer effects, the immune-modulatory effect of Glycine max has not been reported. In the current study, we have discovered that the extract of Glycine max fermented with B. animalis subsp. lactis LDTM 8102 (GFB 8102), could exert immuno-modulatory properties. GFB 8102 treatment increased the production of immune-stimulatory cytokines in RAW264.7 macrophages without any noticeable cytotoxicity. Analysis of the molecular mechanism revealed that GFB 8102 could upregulate MAPK2K and MAPK signaling pathways including ERK, p38, and JNK. GFB 8102 also increased the proliferation rate of splenocytes isolated from mice. In an animal study, administration of GFB 8102 partially recovered cyclophosphamide-mediated reduction in thymus and spleen weight. Moreover, splenocytes from the GFB 8102-treated group exhibited increased TNF-α, IL-6, and IL-1β production. Based on these findings, GFB 8102 could be a promising functional food material for enhancing immune function.


Preservation of hypothalamic dopaminergic neurons in Parkinson's disease.
The integrity of the hypothalamic neuroendocrine dopaminergic neurons in Parkinson's disease was assessed by comparing the numbers and distribution of melanin-pigmented neurons in the arcuate and periventricular nuclei in 7 parkinsonian and 5 normal brains. No significant differences were observed. In contrast to theories suggesting that Parkinson's disease involves a universal degeneration of central catecholaminergic neurons, the hypothalamic neuroendocrine dopaminergic neurons do not appear to be involved by the disease.


[Breast recurrence after conservative treatment of cancers of the breast: predictive factors and prognostic significance].
The prognostic significance of local relapse after conservative treatment of early stage breast cancer has been controversial. To determine the prognostic value of breast relapse occurrence and the factors predisposing to local failure, we analysed the results obtained in a series of 324 patients conservatively treated for breast invasive carcinomas of 2 cm or less. All patients were treated by tumorectomy and axillary dissection, radiation therapy, and adjuvant medical treatment in 85 of 109 patients with involved nodes. The overall 5-yr survival rate was 91.5%. The actuarial 5-yr local control rate was 91.9%. Local relapses occurred in 25 patients. In these patients, the overall 5-yr survival rate was significantly decreased as compared to that of patients without local relapse (76.9% versus 93.4%, P less than 0.001). Recurrence in the breast within 2 yr of initial treatment of severely affected the risk of metastasis occurrence as compared to later relapses (metastasis-free survival rate of 14.3% versus 63.9% at 3-yr from local relapse, P less than 0.01). Local relapse occurred more frequently in premenopausal patients, in patients with low bodymass index, and in patients with small breast size. Local control was not significantly affected by age, tumor size or node involvement. We conclude that the occurrence of a breast relapse after a local conservative treatment has a negative significance and is predictive of a high risk of distant metastasis development, especially when local relapse occurred within 2 yr of initial treatment. There is a need to determine individual factors that could allow discrimination of patients with a high probability of local relapse and subsequent metastasis.


Ecosystem change and human health: implementation economics and policy.
Several recent initiatives such as Planetary Health, EcoHealth and One Health claim that human health depends on flourishing natural ecosystems. However, little has been said about the operational and implementation challenges of health-oriented conservation actions on the ground. We contend that ecological-epidemiological research must be complemented by a form of implementation science that examines: (i) the links between specific conservation actions and the resulting ecological changes, and (ii) how this ecological change impacts human health and well-being, when human behaviours are considered. Drawing on the policy evaluation tradition in public economics, first, we present three examples of recent social science research on conservation interventions that affect human health. These examples are from low- and middle-income countries in the tropics and subtropics. Second, drawing on these examples, we present three propositions related to impact evaluation and non-market valuation that can help guide future multidisciplinary research on conservation and human health. Research guided by these propositions will allow stakeholders to determine how ecosystem-mediated strategies for health promotion compare with more conventional biomedical prevention and treatment strategies for safeguarding health.This article is part of the themed issue 'Conservation, biodiversity and infectious disease: scientific evidence and policy implications'.


A reliable radiochromatographic assay technique for hepatic microsomal 16 alpha-hydroxylase activity towards oestrone 3-sulphate. Comparison between pigmented and non-pigmented mature guinea pigs.
A reliable procedure for the assay of liver microsomal 16 alpha-hydroxylation of oestrone 3-sulphate has been developed for the guinea pig. It is based on the rapid, quantitative separation of oestradiol and oestriol by Sephadex LH-20 columns after the chemical reduction and enzymic hydrolysis of the incubation products. Microsomal preparations and incubation conditions that optimized 16 alpha-hydroxylation of oestrone 3-sulphate were employed. Under these circumstances, reduction of the substrate at C-17 and hydrolysis of the sulphate were minimized. Conditions were established that yielded reaction linearity with respect to time and microsomal concentration. This hydroxylation had an absolute requirement for NADPH, which could not be satisfied by NADH. Apparent Km values for oestrone 3-sulphate and NADPH, under the conditions used, were 14 microM and 0.17 mM respectively. 16 alpha-Hydroxylase activity was present in the liver microsomal fraction from heavily pigmented, female English Shorthaired guinea pigs. Much lower activity was detected in mature pigmented males and albino females. No activity could be demonstrated in mature, albino males.


Examination of mitochondrial protein targeting of haem synthetic enzymes: in vivo identification of three functional haem-responsive motifs in 5-aminolaevulinate synthase.
The initial and the terminal three enzymes of the mammalian haem biosynthetic pathway are nuclear encoded, cytoplasmically synthesized and post-translationally translocated into the mitochondrion. The first enzyme, ALAS (5-aminolaevulinate synthase), occurs as an isoenzyme encoded on different chromosomes and is synthesized either as a housekeeping protein (ALAS-1) in all non-erythroid cell types, or only in differentiating erythroid precursor cells (ALAS-2). Both ALAS proteins possess mitochondrial targeting sequences that have putative haem-binding motifs. In the present study, evidence is presented demonstrating that two haem-binding motifs in the leader sequence, as well as one present in the N-terminus of the mature ALAS-1 function in vivo in the haem-regulated translocation of ALAS-1. Coproporphyrinogen oxidase, the antepenultimate pathway enzyme, possesses a leader sequence that is approx. 120 residues long. In contrast with an earlier report suggesting that only 30 residues were required for translocation of the coproporphyrinogen oxidase, we report that the complete leader is necessary for translocation and that this process is not haem-sensitive in vivo. PPO (protoporphyrinogen oxidase) lacks a typical mitochondrial targeting leader sequence and was found to be effectively targeted by just 17 N-terminal residues. Bacillus subtilis PPO, which is very similar to human PPO at its N-terminal end, is not targeted to the mitochondrion when expressed in mammalian cells, demonstrating that the translocation is highly specific with regard to both the length and spacing of charged residues in this targeting region. Ferrochelatase, the terminal enzyme, possesses a typical N-terminal leader sequence and no evidence of a role for the C-terminus was found in mitochondrial targeting.


Thyroid Cancer Incidence Rates in North Dakota are Associated with Land and Water Use.
Objective: The increasing rate of thyroid cancer diagnoses in the U.S. reflects the increasing use of ultrasonography and of specialist medical care. North Dakota is a rural state with limited access to specialist care, yet its incidence of thyroid cancer is significantly greater than that of the U.S. overall. We sought to identify factors responsible for the high incidence of thyroid cancer in North Dakota. Methods: We examined county-specific incidence rates for thyroid cancer in North Dakota in relation to demographic and geographic factors, including median household income, percent of land fertilized, cattle density per capita, and source of drinking water (city or well water), using structural equation modeling. We included county level data on residential radon levels and estimates of radioactive iodine in milk following nuclear weapons testing in the 1950s. Results: Thyroid cancer incidence rates were significantly associated with median income (p < 0.05); percent of land fertilized (p < 0.05); the use of city water (p < 0.01), and cattle density per capita (p < 0.001). Conclusions: The risk of thyroid cancer in North Dakota is positively associated with income and with factors related to land and water use. Our finding that thyroid cancer incidence rates are associated with the use of city water was unexpected and merits examination in other locations with a mix of city and well water use.


Precision application of aldicarb to enhance efficiency of thrips (Thysanoptera: Thripidae) management in cotton.
Field studies were conducted during 1999-2001 in two climatic/edaphic areas of Georgia (Southern Piedmont and East Gulf Coastal Plain) to test the hypothesis that precision placement of aldicarb with cotton seed in hill planting at spatially specific intervals could decrease insecticide use for management of tobacco thrips, Frankliniella fusca (Hinds). Precision-placed aldicarb controlled thrips during cotton seedling stages using per ha amounts of one-half or less than standard in-furrow application rates with no significant differences in yield. Residual analysis of cotton plants showed that plants in precision placement plots had as much or more aldicarb and aldicarb metabolites present as compared with cotton treated with conventional in-furrow treatments. Higher rates of precision-placed aldicarb did cause phytotoxic burning early in the growing season, but no significant impact on yield was observed.


Paliperidone palmitate, a potential long-acting treatment for patients with schizophrenia. Results of a randomized, double-blind, placebo-controlled efficacy and safety study.
We evaluated the efficacy and safety of the investigational long-acting injectable antipsychotic agent paliperidone palmitate (PP) in the treatment of schizophrenia. Patients were randomized to receive gluteal injections of placebo or PP (50 or 100 mg eq., fixed doses), without oral supplementation, on days 1, 8, and 36 (9-wk, double-blind phase) in this phase 2b study. Patients (n=197, intent-to-treat analysis set) were 62% men, mean (s.d.) age 39 (10) yr, with a baseline mean (s.d.) Positive and Negative Syndrome Scale (PANSS) total score of 87.0 (12.5). Mean (s.d.) PANSS total scores showed significant improvement at endpoint (primary measure) for both the PP 50 mg eq. [-5.2 (21.5)] and PP 100 mg eq. [-7.8 (19.4)] groups, vs. placebo [6.2 (18.3)] (p0.001, each dose vs. placebo). This improvement was detected by day 8 and maintained to endpoint (p0.011) for both doses. In the safety analysis set (n=247), fewer PP-treated patients (2%) discontinued for treatment-emergent adverse events vs. placebo-treated (10%). Rates of treatment-emergent extrapyramidal syndrome-related adverse events were comparable between active treatment and placebo, with the exception of parkinsonism-related disorders (50 mg eq. 5%, 100 mg eq. 8%, placebo 1%). Results of other safety measures suggest PP to be generally well-tolerated. Throughout the study, investigators rated injection-site pain as absent (56-71%), mild (24-39%), moderate (2-12%), or severe (0-2%). PP (50 and 100 mg eq. doses) administered as a gluteal intramuscular injection was efficacious and generally tolerated in these patients with acute symptomatic schizophrenia.


The impact of P-glycoprotein mediated efflux on absorption of 11 sedating and less-sedating antihistamines using Caco-2 monolayers.
Caco-2 cells were used to compare P-gp mediated efflux and passive permeability using bidirectional transport of 11 antihistamines. An efflux ratio >2 indicated active efflux, with PSC833 and GF120918 used as functional P-gp inhibitors. Antihistamines were measured directly by HPLC or LC/MS. Fexofenadine had an efflux ratio of 37, yet had negligible passive permeability, even in the presence of a pH gradient (0.1 × 10(-6) cm/sec). Its precursor, terfenadine, had an efflux ratio of 2.5, while cetirizine, desloratadine and hydroxyzine were 4, 7 and 14, respectively. After incubation with P-gp inhibitors, these ratios dropped significantly. Loratadine, by contrast, had equivalent transport in both directions and passive permeability was high (24 × 10(-6) cm/sec). Dimenhydrinate was the only other sedating antihistamine to exhibit efflux, with a ratio of 10. Gradient conditions of pH (6/7.4) increased efflux of terfenadine and desloratadine to over 31 and 38 fold respectively, yet this increased efflux was not associated with P-gp. Altering functional P-gp in the gut is likely to influence absorption of some sedating antihistamines such as dimenhydrinate and hydroxyzine and most less-sedating antihistamines except loratadine. In addition, desloratadine exhibits pH dependent efflux which could further induce variable absorption of this antihistamine.


A New Class of Selective ATM Inhibitors as Combination Partners of DNA Double-Strand Break Inducing Cancer Therapies.
Radiotherapy and chemical DNA-damaging agents are among the most widely used classes of cancer therapeutics today. Double-strand breaks (DSB) induced by many of these treatments are lethal to cancer cells if left unrepaired. Ataxia telangiectasia-mutated (ATM) kinase plays a key role in the DNA damage response by driving DSB repair and cell-cycle checkpoints to protect cancer cells. Inhibitors of ATM catalytic activity have been shown to suppress DSB DNA repair, block checkpoint controls and enhance the therapeutic effect of radiotherapy and other DSB-inducing modalities. Here, we describe the pharmacological activities of two highly potent and selective ATM inhibitors from a new chemical class, M3541 and M4076. In biochemical assays, they inhibited ATM kinase activity with a sub-nanomolar potency and showed remarkable selectivity against other protein kinases. In cancer cells, the ATM inhibitors suppressed DSB repair, clonogenic cancer cell growth, and potentiated antitumor activity of ionizing radiation in cancer cell lines. Oral administration of M3541 and M4076 to immunodeficient mice bearing human tumor xenografts with a clinically relevant radiotherapy regimen strongly enhanced the antitumor activity, leading to complete tumor regressions. The efficacy correlated with the inhibition of ATM activity and modulation of its downstream targets in the xenograft tissues. In vitro and in vivo experiments demonstrated strong combination potential with PARP and topoisomerase I inhibitors. M4076 is currently under clinical investigation.


Biomechanical Loading Evaluation of Unsintered Hydroxyapatite/poly-l-lactide Plate System in Bilateral Sagittal Split Ramus Osteotomy.
OSTEOTRANS MX® (Takiron Co., Ltd., Osaka, Japan) is a bioactive resorbable maxillofacial osteosynthetic material composed of an unsintered hydroxyapatite/poly-l-lactide composite, and its effective osteoconductive capacity has been previously documented. However, the mechanical strength of this plate system is unclear. Thus, the aim of this in vitro study was to assess its tensile and shear strength and evaluate the biomechanical intensity of different osteosynthesis plate designs after sagittal split ramus osteotomy by simulating masticatory forces in a clinical setting. For tensile and shear strength analyses, three mechanical strength measurement samples were prepared by fixing unsintered hydroxyapatite/poly-l-lactide composed plates to polycarbonate skeletal models. Regarding biomechanical loading evaluation, 12 mandibular replicas were used and divided into four groups for sagittal split ramus osteotomy fixation. Each sample was secured in a jig and subjected to vertical load on the first molar teeth. Regarding shear strength, the novel-shaped unsintered hydroxyapatite/poly-l-lactide plate had significantly high intensity. Upon biomechanical loading evaluation, this plate system also displayed significantly high stability in addition to bioactivity, with no observed plate fracture. Thus, we have clearly demonstrated the efficacy of this plate system using an in vitro model of bilateral sagittal split ramus osteotomy of the mandible.


[T- and B-lymphocyte content in the peripheral blood of healthy subjects].
It was revealed by the method of rosette-formation that the content T- and B-lymphocytes differed in human blood depending on the time of the day and the season. T-cells were at their maximum in the morning and B-cells -- in the evening. The percentage content of B-cells was greater in autumn than in winter.


Identification of the protein coding capability of coronavirus defective viral genomes by mass spectrometry.
During coronavirus infection, in addition to the well-known coronavirus genomes and subgenomic mRNAs, an abundance of defective viral genomes (DVGs) can also be synthesized. In this study, we aimed to examine whether DVGs can encode proteins in infected cells. Nanopore direct RNA sequencing and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis were employed. With the protein databases generated by nanopore direct RNA sequencing and the cell lysates derived from the RNA-protein pull-down assay, six DVG-encoded proteins were identified by LC-MS/MS based on the featured fusion peptides caused by recombination during DVG synthesis. The results suggest that the coronavirus DVGs have the capability to encode proteins. Consequently, future studies determining the biological function of DVG-encoded proteins may contribute to the understanding of their roles in coronavirus pathogenesis and the development of antiviral strategies.


Ultrafast solvation dynamics and charge transfer reactions in room temperature ionic liquids.
Room temperature ionic liquids (ILs) are a new type of solvent with peculiar properties. ILs are usually composed of an anion and a bulky cation with one or more alkyl chains to decrease the melting point. These structural peculiarities lead to the high viscosity and the heterogeneity of ILs, which could affect chemical reactions. In the present perspective, we will first introduce the experimentally observed nature of the heterogeneous liquid structure and then introduce recent developments in the study on electron transfer (ET) and charge transfer (CT) reactions in relation with the solvation and the heterogeneity of ILs. Because of the high viscosity of ILs, diffusive solvation is expected to be slow which could be the rate-limiting factor for ET and CT processes. However, ILs could provide a unique reaction field depending on the location of the solute within the heterogeneous liquid structure and the reaction could be faster than that expected from the bulk viscosity due to the fast fluctuation of the local environment.


Properties of human connexin 31, which is implicated in hereditary dermatological disease and deafness.
The connexins are a family of at least 20 homologous proteins in humans that form aqueous channels connecting the interiors of coupled cells and mediating electrical and chemical communication. Mutations in the gene for human connexin 31 (hCx31) are associated with disorders of the skin and auditory system. Alterations in functional properties of Cx31 junctions are likely to play a role in these diseases; nonetheless, little is known about the properties of the wild-type channels. Here we show that hCx31 channels, like other connexin channels, are gated by voltage and close at low pH and when exposed to long-chain alkanols. Single-channel conductance of the fully open channel is approximately 85 pS, and it is permeable to Lucifer yellow, Alexa Fluor(350), ethidium bromide, and DAPI, which have valences of -2, -1, +1, and +2, respectively. In contrast to what has been reported for mouse Cx31, hCx31 appears to form functional heterotypic channels with all four connexins tested, Cx26, Cx30, Cx32, and Cx45. These findings provide an important first step in evaluating the pathogenesis of inherited human diseases associated with mutations in the gene for Cx31.


Practice effects associated with the repeated assessment of cognitive function using the CogState battery at 10-minute, one week and one month test-retest intervals.
There are many situations in which cognitive tests need to be administered on more than two occasions and at very brief test-retest intervals to detect change in group performance. However, previous literature has not specifically addressed these important issues. The main aim of the current study was to examine these two factors by using a computerized cognitive battery designed specifically for the repeated assessment of cognition (i.e., CogState) in healthy young adult individuals. A further aim of the study was to examine how many times the battery needed to be completed before performance, as measured by the battery, stabilized. Forty-five adults (age range: 18-40 years) completed the battery four times at 10-minute test-retest intervals, and a fifth time at an interval of one week. The results illustrated that when brief test-retest intervals were used (i.e., 10 minutes), performance stabilized after the second assessment, as significant practice effects were generally observed between the first and the second assessments. Practice effects were also observed on some of the tasks at a one-week test-retest interval. Due to these findings, 55 adults (age range: 18-40 years) completed the battery twice at 10-minute test-retest intervals (i.e., to eliminate the initial practice effect), and a third time at an interval of one month. No practice effects were observed. The implications of the results are discussed in terms of methods that can be adopted in order to minimize practice effects when this particular cognitive battery is used.


The challenges of testing for HIV in women: experience from the UK and other European countries.
Strategies for HIV testing vary across Europe, but widespread unacceptably high rates of late diagnosis among women suggest that current testing strategies are not adequately reaching the female population. The only group within the female population that is given specific consideration in guidelines for HIV testing is women who are pregnant, where opt-out testing is often recommended. This article reviews experience from the UK and other European countries with antenatal testing, and trials of routine HIV testing in a range of other settings. Further recommendations for testing strategies for women are required, but need to consider physician barriers to testing, such as constraints on time and training. It is vital to evaluate the motivations that underlie a woman's decision to accept or refuse testing, as these often differ from those of men. Similarly, the consequences of a positive diagnosis for a woman need to be addressed in pre- and post-test discussions.


Cellulose aerogel composites as oil sorbents and their regeneration.
With every oil tanker comes the risk of an accident and oil spill. Sorbents are the most suitable means to remove oil spills. Aerogels as sorbents have high porosity and can be made from cellulose from paper waste. The literature does not distinguish between paper and cardboard as sources of cellulose aerogels and little is known about composites of cellulose aerogels consisting of cellulose fibres and chemically untreated, unprocessed fibres or particles of straw, wool, macroalgae or cellulose acetate from cigarette butts. In this study, the sorption properties for marine diesel oil and biodiesel of such aerogels and their regenerative capacity with bioethanol were investigated. Cellulose aerogels were prepared from office paper and cardboard waste without and with chemically untreated algae, straw, wool and cellulose acetate as a composite by freeze drying. All samples were hydrophobised with methylsilane. The density to calculate the porosity and the contact angle were determined. Then the sorption capacity was determined over five cycles of sorption of oil and regeneration with bioethanol. The average contact angle of all samples was 125°, indicating hydrophobicity. Paper-based aerogels were found to consistently have higher sorption capacities for biodiesel, marine diesel oil and bioethanol than cardboard-based aerogels. In particular, the wool/cellulose aerogel composite was found to have better sorption capacity for biodiesel, marine diesel oil and bioethanol than all other samples. The cellulose acetate/cellulose aerogel composite showed significantly higher sorption capacities than the paper and cardboard control samples (highest value is 32.25 g g-1) only when first used as a sorbent for biodiesel, but with a rapid decrease in the following cycles.


Infarct topography and functional outcomes.
Acute ischemic stroke represents a major cause of long-term adult disability. Accurate prognostication of post-stroke functional outcomes is invaluable in guiding patient care, targeting early rehabilitation efforts, selecting patients for clinical research, and conveying realistic expectations to families. The involvement of specific brain regions by acute ischemia can alter post-stroke recovery potential. Understanding the influences of infarct topography on neurologic outcomes holds significant promise in prognosis of functional recovery. In this review, we discuss the recent evidence of the contribution of infarct location to patient management decisions and functional outcomes after acute ischemic stroke.


Modelling and sensitivity analysis of the reactions involving receptor, G-protein and effector in vertebrate olfactory receptor neurons.
A biochemical model of the receptor, G-protein and effector (RGE) interactions during transduction in the cilia of vertebrate olfactory receptor neurons (ORNs) was developed and calibrated to experimental recordings of cAMP levels and the receptor current (RC). The model describes the steps from odorant binding to activation of the effector enzyme which catalyzes the conversion of ATP to cAMP, and shows how odorant stimulation is amplified and delayed by the RGE transduction cascade. A time-dependent sensitivity analysis was performed on the model. The model output-the cAMP production rate-is particularly sensitive to a few, dominant parameters. During odorant stimulation it depends mainly on the initial density of G-proteins and the catalytic constant for cAMP production.


Creation and Assessment of a Clinical Predictive Calculator and Mortality Associated With Candida krusei Bloodstream Infections.
Candida krusei bloodstream infection (CK BSI) is associated with high mortality, but whether this is due to underlying comorbidities in affected patients or the organism itself is unknown. Identifying patient characteristics that are associated with CK BSI is crucial for clinical decision-making and prognosis. We conducted a retrospective analysis of hospitalized patients with Candida BSI at our institution between 2002 and 2015. Data were collected on demographics, comorbidities, medications, procedures, central lines, vital signs, and laboratory values. Multivariable logistic and Cox regression were used to identify risk factors associated with CK and mortality, respectively. We identified 1873 individual patients who developed Candida BSI within the study period, 59 of whom had CK BSI. CK BSI was predicted by hematologic malignancy, gastric malignancy, neutropenia, and the use of prophylactic azole antifungals, monoclonal antibodies, and β-lactam/β-lactamase inhibitor combinations. The C-statistic was 0.86 (95% confidence interval, 0.81-0.91). The crude mortality rates were 64.4% for CK BSI and 41.4% for non-CK BSI. Although CK was associated with higher mortality in univariable Cox regression, this relationship was no longer significant with the addition of the following confounders: lymphoma, neutropenia, glucocorticoid use, chronic liver disease, and elevated creatinine. Six patient comorbidities predicted the development of CK BSI with high accuracy. Although patients with CK BSI have higher crude mortality rates than patients with non-CK BSI, this difference is not significant when accounting for other patient characteristics.


How Information Drives Complexity.
As systems evolve over time, their natural tendency is to become increasingly more complex. Studies in the field of complex systems have generated new perspectives on the application of management strategies in health systems. This article is the 2nd in a series of articles that focuses on why technological complexity is increasing and strategies nurse administrators can use to successfully implement change in the face of it.


Network pharmacology identification and in vivo validation of key pharmacological pathways of Qin Jiao for gout and arthritis.
Context: Gout is a chronic disease that imposes a huge financial and health burden on patients, which might diminish quality of life. Qin Jiao, a perennial herb found in northwestern China and Japan, is commonly used for treating various ailments.Objective: This study investigates the effects of Qin Jiao on gout and joint inflammation and elucidates its potential mechanism for gouty arthritis.Materials and methods: Study 1, a literature review was conducted using PubMed, Web of Science, and CNKI to assess the applications of Qin Jiao in arthritis treatment. Study 2 was performed to discover the component targets and gouty disease targets via TCMSP, OMIM, GeneCards and DRUGBANK, and network pharmacology analysis. Study 3, male Sprague-Dawley (SD) rats were divided into normal, model, colchicine, Qin Jiao low-dose (QJL), and Qin Jiao high-dose group (QJH), oral gavage for 40 d. Serum, synovial fluid, and synovial membrane tissue were collected to measure the expression levels of IL-1β, IL-6, and STAT3.Results: The research also identified potential targets and pharmacological pathways of Qin Jiao for gout treatment. In vivo study demonstrated Qin Jiao can reduce IL-1β levels in serum and ankle flushing fluid. ELISA analysis confirmed that Qin Jiao significantly reduces the protein expression of IL-6 and STAT3.Discussion and conclusion: Qin Jiao exerts anti-inflammatory effects on gouty arthritis by modulating the IL-6/STAT3 pathway. This study provides a biological basis for the use of Qin Jiao in treating arthritis-related diseases and offers experimental evidence for potential future drug development.


In Vitro Effects of Wistar Rat Prenatal and Postnatal Cerebrospinal Fluid on Neural Differentiation and P roliferation of Mesenchymal Stromal Cells Derived from Bone Marrow.
Cerebrospinal fluid (CSF) plays an important role in cortical development during the fetal stages. Embryonic CSF (E-CSF) consists of numerous neurotrophic and growth factors that regulate neurogenesis, differentiation, and proliferation. Mesenchymal stem cells (MSCs) are multi-potential stem cells that can differentiate into mesenchymal and non-mesenchymal cells, including neural cells. This study evaluates the prenatal and postnatal effects of CSF on proliferation and neural differentiation of bone marrow MSCs (BM-MSCs) at gestational ages E19, E20, and the first day after birth (P1). In this experimental study, we confirmed the mesenchymal nature of BM-MSCs according to their adherence properties and surface markers (CD44, CD73 and CD45). The multi-potential characteristics of BMMSCs were verified by assessments of the osteogenic and adipogenic potentials of these cells. Under appropriate in vitro conditions, the BM-MSCs cultures were incubated with and without additional pre- and postnatal CSF. The MTT assay was used to quantify cellular proliferation and viability. Immunocytochemistry was used to study the expression of MAP-2 and β-III tubulin in the BM-MSCs. We used ImageJ software to measure the length of the neurites in the cultured cells. BM-MSCs differentiated into neuronal cell types when exposed to basic fibroblast growth factor (b-FGF). Viability and proliferation of the BM-MSCs conditioned with E19, E20, and P1 CSF increased compared to the control group. We observed significantly elevated neural differentiation of the BM-MSCS cultured in the CSF-supplemented medium from E19 compared to cultures conditioned with E20 and P1 CSF group. The results have confirmed that E19, E20, and P1 CSF could induce proliferation and differentiation of BM-MSCs though they are age dependent factors. The presented data support a significant, conductive role of CSF components in neuronal survival, proliferation, and differentiation.


Cardiac contractile function during coronary stenosis in dogs: association of adenosine in glycolytic dependence.
We tested the hypothesis that during critical coronary stenosis, endogenous adenosine alters myocardial glucose utilization to support myocardial contractile function (MCF). Anesthetized mongrel dogs were instrumented to measure hemodynamic variables, regional MCF (sonomicrometry), and substrate uptakes. Critical coronary artery stenosis was established with a screw clamp on the left circumflex coronary artery (LCX). Either 8-phenyltheophylline (3 x 10(-7) mol/min; adenosine-receptor blockade), iodoacetate (1 x 10(-5) mol/min; glycolysis blockade), or vehicle was infused into the LCX and the left anterior descending coronary artery (LAD). Critical coronary stenosis caused small decreases in arterial blood pressure and LCX blood flow, but no significant changes in MCF or other hemodynamics. There was a significant decrease in the O2 supply-to-consumption ratio in the stenotic region and an increased glucose uptake. Infusion of either 8-phenyltheophylline or iodoacetate caused a decrease in MCF in the stenotic LCX region concomitant with a decreased glucose uptake and without further changes in blood flow. This was not seen in the nonstenotic (LAD) region. These data support the hypothesis, indicating that glycolysis is vital for maintaining regional MCF during a decrease in the myocardial O2 supply-to-consumption ratio and that adenosine is important in this regard, independent of its vasoactive properties.


Assessing curriculum effectiveness: a survey of Uniformed Services University medical school graduates.
This study assessed alumni perceptions of their preparedness for clinical practice using the Accreditation Council for Graduate Medical Education (ACGME) competencies. We hypothesized that our alumni's perception of preparedness would be highest for military-unique practice and professionalism and lowest for system-based practice and practice-based learning and improvement. 1,189 alumni who graduated from the Uniformed Services University (USU) between 1980 and 2001 completed a survey modeled to assess the ACGME competencies on a 5-point, Likert-type scale. Specifically, self-reports of competencies related to patient care, communication and interpersonal skills, medical knowledge, professionalism, systems-based practice, practice-based learning and improvement, and military-unique practice were evaluated. Consistent with our expectations as the nation's military medical school, our graduates were most confident in their preparedness for military-unique practice, which included items assessing military leadership (M = 4.30, SD = 0.65). USU graduates also indicated being well prepared for the challenges of residency education in the domain of professionalism (M = 4.02, SD = 0.72). Self-reports were also high for competencies related to patient care (M = 3.86, SD = 0.68), communication and interpersonal skills (M = 3.88, SD = 0.66), and medical knowledge (M = 3.78, SD = 0.73). Consistent with expectations, systems-based practice (M = 3.50, SD = 0.70) and practice-based learning and improvement (M = 3.57, SD = 0.62) were the lowest rated competencies, although self-reported preparedness was still quite high. Our findings suggest that, from the perspective of our graduates, USU is providing both an effective military-unique curriculum and is preparing trainees for residency training. Further, these results support the notion that graduates are prepared to lead and to practice medicine in austere environments. Compared to other competencies that were assessed, self-ratings for systems-based practice and practice-based learning and improvement were the lowest, which suggests the need to continue to improve USU education in these areas.


Subdural electrode recording of generalized photoepileptic responses.
We evaluated the spatiotemporal distribution of photic driving (PDR), photoparoxysmal (PPR), and photoconvulsive (PCR) responses recorded by intracranial electrodes (ic-EEG) in a patient with generalized photosensitivity and right frontal lobe cortical dysplasia. Intermittent light stimulation (ILS) was performed thirteen times in nine days. Cortical responses to ILS recorded by ic-EEG were reviewed and classified as PDRs, PPRs, and PCRs. Photic driving responses were restricted to the occipital lobe at ILS frequencies below 9 Hz, spreading to the parietal and central regions at > 9 Hz. Photoparoxysmal responses commonly presented as focal, medial occipital, and parietal interictal epileptic discharges (IEDs), the latter propagating to the sensorimotor cortices. Generalized IEDs were also generated in the setting of PPRs. Photoconvulsive responses, characterized by repetitive bilateral upper extremity myoclonus sustained until the end of the stimulus, were associated with propagation of the medial parieto-occipital discharge to the primary sensorimotor and supplementary area cortices, while generalized myoclonic seizures were associated with a generalized spike-and-wave discharge with an interhemispheric posterior cingulate onset sparing the sensorimotor cortices. Both types of PCR could occur during the same stimulus. Regardless of the pathway, PCRs only occurred when PDRs involved the parietal cortices. While there may be more than one pathway underlying PCRs, parietal lobe association cortices appear to be critical to their generation.


Landscape-scale analyses suggest both nutrient and antipredator advantages to Serengeti herbivore hotspots.
Mechanistic explanations of herbivore spatial distribution have focused largely on either resource-related (bottom-up) or predation-related (top-down) factors. We studied direct and indirect influences on the spatial distributions of Serengeti herbivore hotspots, defined as temporally stable areas inhabited by mixed herds of resident grazers. Remote sensing and variation in landscape features were first used to create a map of the spatial distribution of hotspots, which was tested for accuracy against an independent data set of herbivore observations. Subsequently, we applied structural equation modeling to data on soil fertility and plant quality and quantity across a range of sites. We found that hotspots in Serengeti occur in areas that are relatively flat and located away from rivers, sites where ungulates are less susceptible to predation. Further, hotspots tend to occur in areas where hydrology and rainfall create conditions of relatively low-standing plant biomass, which, coupled with grazing, increases forage quality while decreasing predation risk. Low-standing biomass and higher leaf concentrations of N, Na, and Mg were strong direct predictors of hotspot occurrence. Soil fertility had indirect effects on hotspot occurrence by promoting leaf Na and Mg. The results indicate that landscape features contribute in direct and indirect ways to influence the spatial distribution of hotspots and that the best models incorporated both resource- and predation-related factors. Our study highlights the collective and simultaneous role of bottom-up and top-down factors in determining ungulate spatial distributions.


Asymmetric Lewis acid-catalyzed Diels-Alder reactions of alpha,beta-unsaturated ketones and alpha,beta-unsaturated acid chlorides.
[reaction: see text] Conformational analysis, van der Waals attractions, and transition structure calculations are combined to design an asymmetric Lewis acid-catalyzed Diels-Alder reaction for simple acyclic alpha,beta-unsaturated ketones and alpha,beta-unsaturated acid chlorides, giving up to 83 and 92% ee, respectively. The two-point-binding chiral recognition mechanism, Lewis acid-Lewis base coordination with boron and a van der Waals attraction with the naphthyl group, uses the inherent enone unit of simple alpha,beta-unsaturated carbonyl compounds, ending the need for auxiliary oxygen binding sites on the dienophile.


Genetic analysis of Actinobacillus pleuropneumoniae, and comparison with Haemophilus spp. Taxon "minor group" and Taxon C.
The genetic diversity among 52 field isolates, mainly from Australia, and 15 reference strains of Actinobacillus pleuropneumoniae was examined using multilocus enzyme electrophoresis. Thirty three electrophoretic types (ETs) were recognised, with a mean genetic diversity per locus of 0.312. Australian strains of serovars 1, 2, 5 and 7 belonged to the same clonal lines as strains of these serovars in other countries, but distinct clones of serovars 3, 7, 11 and 12 were also identified. The type strains of serovars 1, 9 and 11 were placed in the same ET. Strains of A. pleuropneumoniae biovar 2 were closely related to biovar 1 strains. Twenty three isolates which did not react with the typing sera or could not be assigned to a single serovar were divided into seven clonal groups. Five strains of Haemophilus sp. Taxon "minor group" which were included in the study were distinct from A. pleuropneumoniae, and were diverse, being divided into at least two major genetic groupings. Two strains of Haemophilus sp. Taxon C were placed in another distinct ET.


An integrated teaching method of gross anatomy and computed tomography radiology.
It is essential for medical students to learn and comprehend human anatomy in three dimensions (3D). With this in mind, a new system was designed in order to integrate anatomical dissections with diagnostic computed tomography (CT) radiology. Cadavers were scanned by CT scanners, and students then consulted the postmortem CT images during cadaver dissection to gain a better understanding of 3D human anatomy and diagnostic radiology. Students used handheld digital imaging and communications in medicine viewers at the bench-side (OsiriX on iPod touch or iPad), which enabled "pixel-to-tissue" direct comparisons of CT images and cadavers. Students had lectures and workshops on diagnostic radiology, and they completed study assignments where they discussed findings in the anatomy laboratory compared with CT radiology findings. This teaching method for gross and radiological anatomy was used beginning in 2009, and it yielded strongly positive student perspectives and significant improvements in radiology skills in later clinical courses.


Influences of aspartate on circadian patterns of lipid peroxidation products and antioxidants in Wistar rats.
The aim of the present study was to demonstrate the effects of aspartate on the circadian patterns of lipid peroxidation products and antioxidants in aspartate-treated rats, in order to investigate the influences of aspartate and whether it could modulate these rhythms differently, since aspartate is an important excitatory neurotransmitter (present in retinohypothalamic tract and suprachiasmatic nuclei [SCN]) involved in the generation and regulation of circadian rhythmicity. Aspartate (50 mg/kg body weight) was administered orally for 60 days to Wistar rats, and 24-hour rhythms of thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase were studied under semi-natural (light/dark 12:12 hr) conditions. Exogenous aspartate administration caused acrophase advances in TBARS rhythms, and delays in GSH, SOD and catalase rhythms; altered MESOR and decreased amplitude values were also seen in all of these rhythms. Our findings indicate that the orally-treated aspartate could reach the hypothalamus, and various brain centres possibly including SCN, and could modulate the circadian patterns of lipid peroxidation products and antioxidants.


Insights into the biology and treatment strategies of pancreatic neuroendocrine tumors.
Pancreatic neuroendocrine tumors (PNETs) are the second most common primary pancreatic neoplasms after pancreatic ductal adenocarcinoma. PNETs present with widely various clinical manifestation and unfavorable survival rate. The recent advances in next generation sequencing have significantly increased our understanding of the molecular landscape of PNETs and help guide the development of targeted therapies. This review intends to outline a holistic picture of the tumors by discussing current understanding of clinical presentations, up-to-date treatment strategies, novel mouse models, and molecular biology of PNETs. Furthermore, we will provide insight into the future development of more effective targeted therapies that are necessary to manage PNETs.


Partner-seeking and limbic dopamine system are enhanced following social loss in male prairie voles (Microtus ochrogaster).
Death of a loved one is recognized as one of life's greatest stresses, and 10%-20% of bereaved individuals will experience a complicated or prolonged grieving period that is characterized by intense yearning for the deceased. The monogamous prairie vole (Microtus ochrogaster) is a rodent species that forms pair bonds between breeding partners and has been used to study the neurobiology of social behaviors and isolation. Male prairie voles do not display distress after isolation from a familiar, same-sex conspecific; however, separation from a bonded female partner increases emotional, stress-related, and proximity-seeking behaviors. Here, we tested the investigatory response of male voles to partner odor during a period of social loss. We found that males who lost their partner spent significantly more time investigating partner odor but not non-partner social odor or food odor. Bachelor males and males in intact pairings did not respond uniquely to any odor. Furthermore, we examined dopamine (DA) receptor mRNA expression in the anterior insula cortex (aIC), nucleus accumbens (NAc), and anterior cingulate (ACC), regions with higher activation in grieving humans. While we found some effects of relationship type on DRD1 and DRD2 expression in some of these regions, loss of a high-quality opposite-sex relationship had a significant effect on DA receptor expression, with pair-bonded/loss males having higher expression in the aIC and ACC compared with pair-bonded/intact and nonbonded/loss males. Together, these data suggest that both relationship type and relationship quality affect reunion-seeking behavior and motivational neurocircuits following social loss of a bonded partner.


Glucocorticoids, 11beta-hydroxysteroid dehydrogenase, and fetal programming.
Epidemiological studies in many distinct human populations have associated low weight or thinness at birth with a substantially increased risk of cardiovascular and metabolic disorders, including hypertension and insulin resistance/type 2 diabetes, in adult life. The concept of fetal "programming" has been advanced to explain this phenomenon. Prenatal glucocorticoid therapy reduces birthweight, and steroids are known to exert long-term organizational effects during specific "windows" of development. Therefore, we hypothesized that fetal overexposure to endogenous glucocorticoids might underpin the link between early life events and later disease. In rats, birthweight is reduced following prenatal exposure to the synthetic glucocorticoid dexamethasone, which readily crosses the placenta, or to carbenoxolone, which inhibits 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), the physiological feto-placental "barrier" to endogenous glucocorticoids. Although the offspring regain the weight deficit by weaning, as adults they exhibit permanent hypertension, hyperglycemia, and increased hypothalamic-pituitary-adrenal axis activity. Moreover, physiological variations in placental 11beta-HSD2 activity near term correlate directly with fetal weight. In humans, 11beta-HSD2 gene mutations produce a low birthweight, and some studies show reduced placental 11beta-HSD2 activity in association with intrauterine growth retardation. Moreover, low birthweight babies have higher plasma cortisol levels throughout adult life, indicating that hypothalamic-pituitary-adrenal axis programming also occurs in humans. The molecular mechanisms of glucocorticoid programming are beginning to be unraveled and involve permanent and tissue-specific changes in the expression of key genes, notably of the glucocorticoid receptor itself. Thus, glucocorticoid programming may explain, in part, the association between fetal events and subsequent disorders in adult life.


Influence of maternal age at delivery and birth order on risk of type 1 diabetes in childhood: prospective population based family study. Bart's-Oxford Family Study Group.
To examine the influence of parental age at delivery and birth order on subsequent risk of childhood diabetes. Prospective population based family study. Area formerly administered by the Oxford Regional Health Authority. 1375 families in which one child or more had diabetes. Of 3221 offspring, 1431 had diabetes (median age at diagnosis 10.5 years, range 0.4-28.5) and 1790 remained non-diabetic at a median age of 16. 1 years. Disease free survival and hazard ratios for the development of type 1 diabetes in all offspring, assessed by Cox proportional hazard regression. Maternal age at delivery was strongly related to risk of type 1 diabetes in the offspring; risk increased by 25% (95% confidence interval 17% to 34%) for each five year band of maternal age, so that maternal age at delivery of 45 years or more was associated with a relative risk of 3.11 (2.07 to 4.66) compared with a maternal age of less than 20 years. Paternal age was also associated with a 9% (3% to 16%) increase for each five year increase in paternal age. The relative risk of diabetes, adjusted for parental age at delivery and sex of offspring, decreased with increasing birth order; the overall effect was a 15% risk reduction (10% to 21%) per child born. A strong association was found between increasing maternal age at delivery and risk of diabetes in the child. Risk was highest in firstborn children and decreased progressively with higher birth order. The fetal environment seems to have a strong influence on risk of type 1 diabetes in the child. The increase in maternal age at delivery in the United Kingdom over the past two decades could partly account for the increase in incidence of childhood diabetes over this period.


Automatic Calibration of the Step Length Model of a Pocket INS by Means of a Foot Inertial Sensor.
All non-foot-mounted inertial localization systems have a common challenge: the need for calibrating the parameters of the step length model. The calibration of the parameters of a step length model is key for an accurate estimation of the pedestrian's step length, and therefore, for the accuracy of the position estimation. In a previous work, we provided a proof of concept on how to calibrate step length models with a foot inertial navigation system (INS), i.e., an INS based on an inertial measurement unit (IMU) mounted on the upper front part of the foot. The reason is that the foot INS does not require calibration thanks to the implementation of the strapdown algorithm. The goal of this article is to automatically calibrate the parameters of a step length model of the pocket INS by means of the foot INS. The step length model of the pocket INS has two parameters: the slope and offset of a first-order linear regression that relates the amplitude of the thigh pitch with the user's step length. Firstly, we show that it is necessary to estimate the two parameters of the step length model. Secondly, we propose a method to automatically estimate these parameters by means of a foot INS. Finally, we propose a practical implementation of the proposed method in the pocket INS. We evaluate the pocket INS with the proposed calibration method and we compare the results to the state of the art implementations of the pocket INS. The results show that the proposed automatic calibration method outperforms the previous work, which proves the need for calibrating all the parameters of the step length model of the pocket INS. In this work, we conclude that it is possible to use a foot INS to automatically calibrate all parameters of the step length model of the pocket INS. Since the calibration of the step length model is always needed, our proposed automatic calibration method is a key enabler for using the pocket INS.


[Use of glucocorticoid preparations in congenital adrenocortical dysfunction].
Circadian changes in the corticosteroid, testosterone, and ACTH content in the blood plasma were examined to monitor the suppressant effect of glucocorticoids on adrenocortical and hypophyseal functions during administering glucocorticoid drugs to patients with congenital adrenocortical dysfunction. The daily dose of glucocorticoids was given in two intakes according to the two schemes: scheme 1 included the drug intake in the morning and in the afternoon, scheme 2 in the morning and in the evening. The effect of glucocorticoid therapy on the pattern of the circadian rhythm and the absolute corticosteroid and testosterone content was shown to depend on the time of the drug intake. The treatment according to scheme 2 is preferable: the absolute testosterone content was reducing to normal during the whole day. Meanwhile the normal circadian rhythm of the adrenal cortex and hypophysis was preserved.


Conflicting objectives in regulating the automobile.
Federal regulation of automobile safety, emissions, and fuel economy is contradictory. Safety equipment and emissions control reduce fuel economy; reducing the size of automobiles is estimated to increase fatalities by 1400 a year and significantly increase serious injuries. These secondary impacts of regulation roughly double the estimated costs of achieving the individual goals. In formulating regulations, these contradictions must be taken into account, along with the effects on the price of the vehicle and its attractiveness to buyers.


Gait analysis comparison between manual marking, 2D pose estimation algorithms, and 3D marker-based system.
Recent advances in Artificial Intelligence (AI) and Computer Vision (CV) have led to automated pose estimation algorithms using simple 2D videos. This has created the potential to perform kinematic measurements without the need for specialized, and often expensive, equipment. Even though there's a growing body of literature on the development and validation of such algorithms for practical use, they haven't been adopted by health professionals. As a result, manual video annotation tools remain pretty common. Part of the reason is that the pose estimation modules can be erratic, producing errors that are difficult to rectify. Because of that, health professionals prefer the use of tried and true methods despite the time and cost savings pose estimation can offer. In this work, the gait cycle of a sample of the elderly population on a split-belt treadmill is examined. The Openpose (OP) and Mediapipe (MP) AI pose estimation algorithms are compared to joint kinematics from a marker-based 3D motion capture system (Vicon), as well as from a video annotation tool designed for biomechanics (Kinovea). Bland-Altman (B-A) graphs and Statistical Parametric Mapping (SPM) are used to identify regions of statistically significant difference. Results showed that pose estimation can achieve motion tracking comparable to marker-based systems but struggle to identify joints that exhibit small, but crucial motion. Joints such as the ankle, can suffer from misidentification of their anatomical landmarks. Manual tools don't have that problem, but the user will introduce a static offset across the measurements. It is proposed that an AI-powered video annotation tool that allows the user to correct errors would bring the benefits of pose estimation to professionals at a low cost.


The effect of pre-operative methylprednisolone on early endothelial damage after total knee arthroplasty: a randomised, double-blind, placebo-controlled trial.
We wished to evaluate whether inhibition of the systemic inflammatory response by a single pre-operative dose of methylprednisolone reduced markers of early endothelial damage after fast-track total knee arthroplasty. We randomly allocated 70 patients undergoing elective unilateral total knee arthroplasty (1:1) to receive either pre-operative intravenous methylprednisolone 125 mg (methylprednisolone group) or isotonic saline (control group). All procedures were performed under spinal anaesthesia without a tourniquet, using a standardised multimodal analgesic regime. The outcomes included changes in Syndecan-1 concentrations, a marker of glycocalyx degradation, markers of endothelial cell damage and activation (plasma soluble thrombomodulin and sE-Selectin), and permeability by vascular endothelial growth factor, as well as C-reactive protein concentrations. Blood samples were collected at baseline and 2 h, 6 h and 24 h after surgery, with complete sampling from 63 patients for analyses. Methylprednisolone significantly reduced markers of endothelial damage at 24 h following surgery compared with saline (methylprednisolone group vs. control group, adjusted means (SEM)) expressed by circulating Syndecan-1: 11.6 (1.0) ng.ml-1 vs. 13.4 (1.1) ng.ml-1 p = 0.046; soluble thrombomodulin: 5.1 (0.1) ng.ml-1 vs. 5.7 (0.2) ng.ml-1 , p = 0.009; sE-Selectin: 64.8 (1.8) ng.ml-1 vs. 75.7 (1.9) ng.ml-1 , p = 0.001, and vascular endothelial growth factor: 35.3 (2.7) ng.ml-1 vs. 58.5 (2.8) ng.ml-1 , p < 0.001. The effect of the intervention increased with time for soluble thrombomodulin, sE-Selectin and vascular endothelial growth factor, and was more pronounced in patients with high baseline values. Finally, methylprednisolone reduced the C-reactive protein response 24 h postoperatively; 31.1 (1.1) mg.l-1 vs. 68.4 (1.1) mg.l-1 , p < 0.001. Pre-operative administration of methylprednisolone 125 mg reduced circulating markers of endothelial activation and damage, as well as the systemic inflammatory response (C-reactive protein) early after fast-track total knee arthroplasty. These findings may have a positive effect on surgical outcome, but require studies in major surgery.


Physical activity consultation for people with Type 2 diabetes: evidence and guidelines.
Physical activity is an important, but often underused, therapeutic strategy within diabetes care. To date, little is known about the best way to promote physical activity in diabetes care. Physical activity consultation is an intervention designed to promote physical activity behaviour change. This article provides guidelines on how to conduct a physical activity consultation with people who have Type 2 diabetes, and reviews the evidence surrounding the effectiveness of this intervention in this population. The trans-theoretical model is the underlying theory of behaviour change for the physical activity consultation intervention. The review identifies research which supports the use of this model for understanding physical activity behaviour in people with Type 2 diabetes. The review outlines a number of modifiable variables associated with physical activity behaviour change in this population. How each of these variables is addressed within the guidelines for conducting a physical activity consultation is identified. Finally, limited but consistent research highlights the effectiveness of physical activity consultation for promoting physical activity in people with Type 2 diabetes.


Chronic Eyelid Edema Following Periocular Hyaluronic Acid Filler Treatment.
A 54-year-old woman received multiple injections of hyaluronic acid filler to the brow region to address volume loss over a 21-month period. She then developed significant pitting edema of both upper eyelids, which persisted for 6 years. Hyaluronidase (Hyalase) was injected into the subcutaneous brows and resulted in complete resolution of the edema within 2 days. This confirms that the hyaluronic acid injected into the brows was responsible for this patient's chronic eyelid edema. This case illustrates an unusual long-term complication of periocular hyaluronic acid filler.


Local infiltration and spermatic cord block for inguinal, scrotal and testicular surgery.
This was a prospective study involving 372 male patients. Surgical procedures including simple inguinal hernia repair, inguinal lymph node biopsy, hydrocelectomy, testicular biopsy, testicular fixation, orchidectomy and scrotal exploration were performed under local anaesthesia using various quantities of 0.5% xylocaine with adrenaline depending on the procedure, in the form of spermatic cord block and local infiltration nerve blocks. No premedication was given to any patient and only five patients (1.34%) were given intraoperative sedation due to anxiety. No complication directly attributed to the anaesthetic agent used or the technique of spermatic cord and nerve blocks were reported during the study. Three hundred and sixty patients (96.77%) were operated on as outpatients and were happy and satisfied to return home on the same day. This experience confirms that spermatic cord block accompanied by local infiltration with 0.5% xylocaine with adrenaline is simple, safe and effective technique that should be used more widely in outpatient urological and general surgical settings in this locality. It provides excellent intra-scrotal and inguinal anaesthesia. Furthermore, the technique is cost effective, and personnel effective since no anaesthetist is required for the procedure which is usually carried out by the surgeon. This would enable many more people to afford the surgical procedures.


Effects of ferric carboxymaltose on hospitalisations and mortality rates in iron-deficient heart failure patients: an individual patient data meta-analysis.
Iron deficiency (ID) is a common co-morbidity in patients with heart failure (HF) and has been suggested to be associated with poor prognosis. Recently completed double-blind randomised controlled trials (RCTs) studying HF patients with ID have shown improvements in functional capacity, symptoms and quality of life when treated with i.v. ferric carboxymaltose (FCM). This individual patient data meta-analysis investigates the effect of FCM vs. placebo on recurrent hospitalisations and mortality in HF patients with ID. Individual patient data were extracted from four RCTs comparing FCM with placebo in patients with systolic HF and ID. The main outcome measures were recurrent cardiovascular (CV) hospitalisations and CV mortality. Other outcomes included cause-specific hospitalisations and death. The main analyses of recurrent events were backed up by time-to-first-event analyses. In total, 839 patients, of whom 504 were randomised to FCM, were included. Compared with those taking placebo, patients on FCM had lower rates of recurrent CV hospitalisations and CV mortality [rate ratio 0.59, 95% confidence interval (CI) 0.40-0.88; P = 0.009]. Treatment with FCM also reduced recurrent HF hospitalisations and CV mortality (rate ratio 0.53, 95% CI 0.33-0.86; P = 0.011) and recurrent CV hospitalisations and all-cause mortality (rate ratio 0.60, 95% CI 0.41-0.88; P = 0.009). Time-to-first-event analyses showed similar findings, with somewhat attenuated treatment effects. The administration of i.v. FCM was not associated with an increased risk for adverse events. Treatment with i.v. FCM was associated with a reduction in recurrent CV hospitalisations in systolic HF patients with ID.


Modeling cell population dynamics.
Quantitative modeling is quickly becoming an integral part of biology, due to the ability of mathematical models and computer simulations to generate insights and predict the behavior of living systems. Single-cell models can be incapable or misleading for inferring population dynamics, as they do not consider the interactions between cells via metabolites or physical contact, nor do they consider competition for limited resources such as nutrients or space. Here we examine methods that are commonly used to model and simulate cell populations. First, we cover simple models where analytic solutions are available, and then move on to more complex scenarios where computational methods are required. Overall, we present a summary of mathematical models used to describe cell population dynamics, which may aid future model development and highlights the importance of population modeling in biology.


Celiac disease: similar presentations in the elderly and young adults.
Studies have shown that celiac disease can affect individuals in all age groups. However, few studies have described the disease in the elderly. The goal of this study is to characterize celiac disease in the elderly by comparing to a population of young adults with celiac disease. Review of a tertiary center database of patients with celiac disease was performed to identify two groups of patients, an elderly cohort ≥ 65 years and a young adult cohort aged 18-30 years, with biopsy-confirmed celiac disease. Information obtained included symptom duration, clinical presentation, small intestinal pathology, associated conditions, and the presence of bone disease. Included in the study were 149 young adult and 125 elderly patients; the latter represented 12.4% of the patients in our database. The duration of symptoms prior to diagnosis was similar, 5.8 ± 12 years and 6.14 ± 12.6 years in the young adult and elderly cohorts, respectively (p = 0.119). There was no significant difference in the mode of presentation of illness. Diarrhea was the main presenting symptom (49% in young adults vs. 50% in the elderly, p = 0.921). There was a similar prevalence of autoimmune disease (19% in young adults vs. 26% in the elderly, p = 0.133). Thyroid disease and neuropathy were more prevalent in the elderly (p = 0.037 and p = 0.023, respectively). The degree of villous atrophy and prevalence of bone disease were similar in each group. Surprisingly, the presentation of celiac disease both clinically and histologically is similar in elderly and young adult patients. The factors triggering disease at any given age remain unclear and warrant further study.


Microbial and mineralogical characterizations of soils collected from the deep biosphere of the former Homestake gold mine, South Dakota.
A microbial census on deep biosphere (1.34 km depth) microbial communities was performed in two soil samples collected from the Ross and number 6 Winze sites of the former Homestake gold mine, Lead, South Dakota using high-density 16S microarrays (PhyloChip). Soil mineralogical characterization was carried out using X-ray diffraction, X-ray photoelectron, and Mössbauer spectroscopic techniques which demonstrated silicates and iron minerals (phyllosilicates and clays) in both samples. Microarray data revealed extensive bacterial diversity in soils and detected the largest number of taxa in Proteobacteria phylum followed by Firmicutes and Actinobacteria. The archael communities in the deep gold mine environments were less diverse and belonged to phyla Euryarchaeota and Crenarchaeota. Both the samples showed remarkable similarities in microbial communities (1,360 common OTUs) despite distinct geochemical characteristics. Fifty-seven phylotypes could not be classified even at phylum level representing a hitherto unidentified diversity in deep biosphere. PhyloChip data also suggested considerable metabolic diversity by capturing several physiological groups such as sulfur-oxidizer, ammonia-oxidizers, iron-oxidizers, methane-oxidizers, and sulfate-reducers in both samples. High-density microarrays revealed the greatest prokaryotic diversity ever reported from deep subsurface habitat of gold mines.


Functions of MutLalpha, replication protein A (RPA), and HMGB1 in 5'-directed mismatch repair.
A purified system comprised of MutSalpha, MutLalpha, exonuclease 1 (Exo1), and replication protein A (RPA) (in the absence or presence of HMGB1) supports 5'-directed mismatch-provoked excision that terminates after mismatch removal. MutLalpha is not essential for this reaction but enhances excision termination, although the basis of this effect has been uncertain. One model attributes the primary termination function in this system to RPA, with MutLalpha functioning in a secondary capacity by suppressing Exo1 hydrolysis of mismatch-free DNA (Genschel, J., and Modrich, P. (2003) Mol. Cell 12, 1077-1086). A second invokes MutLalpha as the primary effector of excision termination (Zhang, Y., Yuan, F., Presnell, S. R., Tian, K., Gao, Y., Tomkinson, A. E., Gu, L., and Li, G. M. (2005) Cell 122, 693-705). In the latter model, RPA provides a secondary termination function, but together with HMGB1, also participates in earlier steps of the reaction. To distinguish between these models, we have reanalyzed the functions of MutLalpha, RPA, and HMGB1 in 5'-directed mismatch-provoked excision using purified components as well as mammalian cell extracts. Analysis of extracts derived from A2780/AD cells, which are devoid of MutLalpha but nevertheless support 5'-directed mismatch repair, has demonstrated that 5'-directed excision terminates normally in the absence of MutLalpha. Experiments using purified components confirm a primary role for RPA in terminating excision by MutSalpha-activated Exo1 but are inconsistent with direct participation of MutLalpha in this process. While HMGB1 attenuates excision by activated Exo1, this effect is distinct from that mediated by RPA. Assay of extracts derived from HMGB1(+/+) and HMGB1(-/-) mouse embryo fibroblast cells indicates that HMGB1 is not essential for mismatch repair.


Effect of heat treatment on results for biochemical analysis of plasma and serum.
Incubation of serum and plasma samples at 56 degrees C for 30 min inactivates the HTLV-III virus. We assessed the effect of this procedure on results of routine biochemical tests by dividing samples of serum and plasma into two, heat-treating one while the other remained at room temperature. Samples were then analyzed in an SMA 16/60, an Astra-8, an Analox glucose analyzer, a Cobas Bio centrifugal analyzer, and manually for salicylate and acetaminophen (paracetamol). Most of the differences produced by heat treatment were not clinically significant, although heated samples proved unsuitable for use in assay of some commonly measured enzymes. Serum evidently is preferable to plasma for this procedure, and heat-treated serum samples can validly be used for most routine analyses. Thus this procedure makes safer the analysis of samples from patients with suspected or proven acquired immune deficiency syndrome (AIDS).


Proton Transfer in Guanine-Cytosine Base Pairs in B-DNA.
A double proton transfer reaction in a guanine-cytosine (GC) base pair has been proposed as a possible mechanism for rare tautomer (G*C*) formation and thus a source of spontaneous mutations. We analyze this system with free energy calculations based on extensive Quantum Mechanics/Molecular Mechanics simulations to properly consider the influence of the DNA biomolecular environment. We find that, although the G*C* rare tautomer is metastable in the gas phase, it is completely unstable in the conditions found in cells. Thus, our calculations show that a double proton reaction cannot be the source of spontaneous point mutations. We have also analyzed the intrabase H transfer reactions in guanine. Our results show that the DNA environment gives rise to a large free energy difference between the rare and canonical tautomers. These results show the key role of the DNA biological environment for the stability of the genetic code.