Pharmacologic basis for high-dose oral acyclovir prophylaxis of cytomegalovirus disease in renal allograft recipients.
The incidence of cytomegalovirus disease, the most important infectious complication of renal transplantation, was reduced in renal allograft recipients by a regimen of prophylactic high-dose oral acyclovir. To analyze the pharmacologic aspects of our prophylactic approach, we evaluated safety, pharmacodynamics, and in vitro susceptibility data. One hundred four recipients of cadaveric renal allografts received either oral acyclovir (n = 53) in doses of up to 3,200 mg/day or a placebo (n = 51) for 12 weeks posttransplant. Leukocyte count and serum creatinine were selected as markers of laboratory safety and were evaluated pretransplant, at study midpoint (creatinine only), and at study completion. Concentrations of acyclovir in plasma were determined to verify the ability of the dosing strategy to achieve predicted values. Viral resistance was assessed by calculation of in vitro 50% inhibitory concentrations (IC50s) of acyclovir for the cytomegalovirus strains collected from the subjects. Our results showed no difference in leukocyte count or serum creatinine between the acyclovir and placebo recipients. Plasma acyclovir concentrations were maintained within the expected limits and did not differ between patients who developed cytomegalovirus disease and those who did not. The mean acyclovir IC50s for cytomegalovirus isolates were 42.6 mumol/liter in the acyclovir recipients and 48 mumol/liter in the placebo recipients. We conclude that the clinical benefit of high-dose oral acyclovir therapy occurred despite plasma drug concentrations below the mean IC50 for the patient viral isolates. Furthermore, the use of the regimen did not produce leukopenia, adversely affect renal function, or alter the susceptibility of cytomegalovirus strains to acyclovir. This approach and dose adjustment scheme may be appropriate for other immunocompromised patients at risk for cytomegalovirus infection and disease.


The effects of naris occlusion on mouse nasal turbinate development.
Unilateral naris occlusion, a standard method for causing odor deprivation, also alters airflow on both sides of the nasal cavity. We reasoned that manipulating airflow by occlusion could affect nasal turbinate development given the ubiquitous role of environmental stimuli in ontogenesis. To test this hypothesis, newborn mice received unilateral occlusion or sham surgery and were allowed to reach adulthood. Morphological measurements were then made of paraffin sections of the whole nasal cavity. Occlusion significantly affected the size, shape and position of turbinates. In particular, the nasoturbinate, the focus of our quantitative analysis, had a more delicate appearance on the occluded side relative to the open side. Occlusion also caused an increase in the width of the dorsal meatus within the non-occluded and occluded nasal fossae, compared with controls, and the position of most turbinates was altered. These results suggest that a mechanical stimulus from respiratory airflow is necessary for the normal morphological development of turbinates. To explore this idea, we estimated the mechanical forces on turbinates caused by airflow during normal respiration that would be absent as a result of occlusion. Magnetic resonance imaging scans were used to construct a three-dimensional model of the mouse nasal cavity that provided the input for a computational fluid dynamics simulation of nasal airflow. The simulation revealed maximum shear stress values for the walls of turbinates in the 1 Pa range, a magnitude that causes remodeling in other biological tissues. These observations raise the intriguing possibility that nasal turbinates develop partly under the control of respiratory mechanical forces.


Suppression of NF-kappaB-mediated beta-defensin gene expression in the mammalian airway by the Bordetella type III secretion system.
Expression of innate immune genes such as beta-defensins is induced in airway epithelium by bacterial components via activation of NF-kappaB. We show here that live Gram-negative bacteria can similarly stimulate this pathway, resulting in upregulation of the beta-defensin tracheal antimicrobial peptide (TAP) in primary cultures of bovine tracheal epithelial cells (TECs), by a Toll-like receptor 4 (TLR4)-mediated pathway. The Gram-negative airway pathogen Bordetella bronchiseptica possesses a type III secretion system previously suggested to inhibit the nuclear translocation of NF-kappaB in a cell line by immunohistochemistry. We therefore hypothesized that this pathogen might interfere in the innate immune response of the epithelium. Exposure of TECs to wild-type B. bronchiseptica suppressed the activation of NF-kappaB and the subsequent induction of TAP mRNA levels, whereas a type III secretion-defective strain did not. These results suggest a mechanism for bacterial evasion of the innate immune response in the airway, which could allow for the observed persistent colonization of this pathogen.


[Stereometric study of the relation between the surfaces of the cardiomyocytes and the walls of cardiac capillaries under normal conditions and in acute myocardial infarct].
Stereometrical analysis of histological myocardial sections stained by the Mac -- Manus method modified by the authors showed a wide individual variation of values characterizing the overall surface and the relative specific surface of the muscle fibers, and the correlation between the relative surfaces of the capillaries and the muscle fibers in the hearts of practically healthy individuals who died in accidents. The myocardium of the anterior papillary muscle and intramural part of the left-ventricular wall were found to have the lowest values. Myocardial infarction occurred rather more frequently in individuals whose heart was characterized by low value of the relationship of the surfaces of the capillary walls and those of the cardiac myocytes and low value of the relative specific surface of the myocardial muscle fibers. This allows such individual structural organization of the myocardium to be accepted as a risk factor of ischemic heart disease and to be considered a manifestation of an unfavourable morphological background promoting earlier ischemia of the myocardium.


Effect of a single high loading dose of rosuvastatin on percutaneous coronary intervention for acute coronary syndromes.
A high loading dose of atorvastatin has been confirmed to reduce postprocedural events in patients undergoing percutaneous coronary intervention (PCI). In this study, we sought to investigate the protective effects of rosuvastatin in patients with acute coronary syndromes (ACS) undergoing PCI and to determine the effect of rosuvastatin pretreatment on the postprocedural levels of high-sensitivity C-reactive protein (hs-CRP), interleukin 6 (IL-6), and monocyte chemotactic protein 1 (MCP-1). A total of 125 patients with non-ST-segment elevation ACS were randomized to pretreatment with rosuvastatin (20 mg 2-4 hours before PCI [n = 62]) or placebo (n = 63). All the patients received subsequent long-term rosuvastatin treatment (10 mg/d). The main end point of the trial was the 30-day incidence of major adverse cardiac events (death, myocardial infarction, or unplanned revascularization). Plasma levels of hs-CRP, IL-6, and MCP-1 were detected before PCI and 6 hours, 24 hours, and 3 days after PCI. The primary end point occurred in 8.1% of the patients in the rosuvastatin arm and 22.2% in the placebo arm (P < .01); this difference was entirely attributed to a reduced incidence of myocardial infarction (8.1% vs 22.2%; P < .01). The postprocedural elevation in creatine kinase-MB and troponin I was also significantly lower in the rosuvastatin group at 6 hours, 24 hours, and 3 days. Plasma levels of hs-CRP, IL-6, and MCP-1 increased significantly after PCI in both the rosuvastatin and control groups; however, the postprocedural elevations in hs-CRP and IL-6 levels were significantly lower in the rosuvastatin group than the control group. A single, high dose (20 mg) of rosuvastatin prior to PCI reduces postprocedural myocardial injury in patients with ACS, with a concomitant attenuation of the postprocedural increase in hs-CRP and IL-6 levels.


Arterial blood pressure measurement and pulse wave analysis--their role in enhancing cardiovascular assessment.
The most common method of clinical measurement of arterial blood pressure is by means of the cuff sphygmomanometer. This instrument has provided fundamental quantitative information on arterial pressure in individual subjects and in populations and facilitated estimation of cardiovascular risk related to levels of blood pressure obtained from the brachial cuff. Although the measurement is taken in a peripheral limb, the values are generally assumed to reflect the pressure throughout the arterial tree in large conduit arteries. Since the arterial pressure pulse becomes modified as it travels away from the heart towards the periphery, this is generally true for mean and diastolic pressure, but not for systolic pressure, and so pulse pressure. The relationship between central and peripheral pulse pressure depends on propagation characteristics of arteries. Hence, while the sphygmomanometer gives values of two single points on the pressure wave (systolic and diastolic pressure), there is additional information that can be obtained from the time-varying pulse waveform that enables an improved quantification of the systolic load on the heart and other central organs. This topical review will assess techniques of pressure measurement that relate to the use of the cuff sphygmomanometer and to the non-invasive registration and analysis of the peripheral and central arterial pressure waveform. Improved assessment of cardiovascular function in relation to treatment and management of high blood pressure will result from future developments in the indirect measurement of arterial blood pressure that involve the conventional cuff sphygmomanometer with the addition of information derived from the peripheral arterial pulse.


PdCl2-catalyzed synthesis of a new class of isocoumarin derivatives containing aminosulfonyl / aminocarboxamide moiety: First identification of a isocoumarin based PDE4 inhibitor.
While anti-inflammatory properties of isocoumarins are known their PDE4 inhibitory potential was not explored previously. In our effort the non-PDE4 inhibitor isocoumarins were transformed into the promising inhibitors via introducing an aminosulfonyl/aminocarboxamide moiety to the C-3 benzene ring attached to the isocoumarin framework. This new class of isocoumarins were synthesized via a PdCl2-catalyzed construction of the 4-allyl substituted 3-aryl isocoumarin ring starting from the appropriate 2-alkynyl benzamide derivative. Several compounds showed good inhibition of PDE4B in vitro and the SAR indicated superiority of aminosulfonamide moiety over aminocarboxamide in terms of PDE4B inhibition. Two compounds 3q and 3u with PDE4B IC50 = 0.43 ± 0.11 and 0.54 ± 0.19 μM and ≥ 2-fold selectivity over PDE4D emerged as initial hits. The participation of aminosulfonamide moiety in PDE4B inhibition and the reason for selectivity though moderate shown by 3q and 3u was revealed by the in silico docking studies. In view of potential usefulness of moderately selective PDE4B inhibitors the compound 3u (that showed PDE4 selectivity over other PDEs) was further evaluated in adjuvant induced arthritic rats. At an intraperitoneal dose of 30 mg/kg the compound showed a significant reduction in paw swelling (in a dose dependent manner), inflammation and pannus formation (in the knee joints) as well as pro-inflammatory gene expression/mRNA levels and increase in body weight. Moreover, besides its TNF-α inhibition and no significant toxicity in an MTT assay the compound did not show any adverse effects in a thorough toxicity studies e.g. teratogenicity, hepatotoxicity, cardiotoxicity and apoptosis in zebrafish. Thus, the isocoumarin 3u emerged as a new, safe and moderately selective PDE4B inhibitor could be useful for inflammatory diseases possibly including COVID-19.


Antimicrobial Activity of Red Alga Flour (Gelidium sp.) and Its Effect on Quality Retention of Scomber scombrus during Refrigerated Storage.
This study analyzed the antimicrobial effect of aqueous extracts of flour obtained from red alga (Gelidium sp.) both in vitro, against most common food pathogenic and spoilage bacteria, and in a food model system during the chilled storage of Atlantic mackerel (Scomber scombrus). Results of in vitro assays allowed the conclusion that the aqueous flour extracts have antimicrobial activity against Gram-negative bacteria such as Enterobacteriaceae (Escherichia coli, Enterobacter aerogenes, and Klebsiella pneumoniae) and proteobacteria (Vibrio alginolyticus), and against Gram-positive bacteria such as Bacillus cereus and B. subtilis. In the food model study, different concentrations of the flour extract were present in the icing medium, microbial and chemical analyses being carried out in fish muscle at different storage times. An inhibitory effect (p < 0.05) on microbial growth (aerobes, psychrotrophs, Enterobacteriaceae, and proteolytic and lipolytic bacteria) and on chemical quality indices (pH, total volatile amines, and trimethylamine) was concluded. This effect was more pronounced when the flour extract concentration in the ice increased and at advanced storage times. This study provides a first approach to the beneficial use of flour of the alga Gelidium as a new preserving strategy for chilled fish.


Synaptic connections of intracellularly filled clutch cells: a type of small basket cell in the visual cortex of the cat.
Light and electron microscopic quantitative analysis was carried out on a type of neuron intracellularly filled with horseradish peroxidase. Two cells were studied in area 17, one of which was injected intra-axonally, and its soma was not recovered. One cell was studied in area 18. The two somata were on the border of layers IVa/b; they were radially elongated and received synapses from numerous large boutons with round synaptic vesicles. The dendrites were smooth and remained largely in layer IV. The cells can be recognised on the basis of their axonal arbor, which was restricted to layer IV (90-95% of boutons) with minor projections to layers III, V, and VI. Many of the large, bulbous boutons contacted neuronal somata, short collaterals often forming "claw"-like configurations around cells. The name "clutch cell" is suggested to delineate this type of neuron from other aspiny multipolar cells. Computer-assisted reconstruction of the axon showed that in layer IV the axons occupied a rectangular area about 300 X 500 microns, elongated anteroposteriorly in area 17 and mediolaterally in area 18. The distributions of synaptic boutons and postsynaptic cells were patchy within this area. A total of 321 boutons were serially sectioned in area 17. The boutons formed type II synaptic contacts. The postsynaptic targets were somata (20-30%), dendritic shafts (35-50%), spines (30%), and rarely axon initial segments. Most of the postsynaptic somata tested were not immunoreactive for GABA and their fine structural features suggest that they are spiny stellate, star pyramidal, and pyramidal neurons. The characteristics of most of the postsynaptic dendrites and spines also suggest that they belong to these spiny neurons. A few of the postsynaptic dendrites and somata exhibited characteristics of cells with smooth dendrites and these somata were immunoreactive for GABA. It is suggested that clutch cells are inhibitory interneurons exerting their effect mainly on layer IV spiny neurons in an area localised perhaps to a single ocular dominance column. The specific laminar location of the axons of clutch cell also suggests that they may be associated with the afferent terminals of lateral geniculate nucleus cells, and could thus be responsible for generating some of the selective properties of neurons of the first stage of cortical processing.


Ability of Thermal Work Limit (TWL) to Assess Sustainable Heat Stress Exposures.
Thermal Work Limit (TWL) recommends a maximum metabolic rate for a given environmental condition, clothing ensemble, and acclimatization state so that thermal equilibrium can be sustained at or below the limiting metabolic rate. The purpose of this paper was to assess the ability of TWL to recommend maximum sustainable levels of heat stress using an existing database of progressive heat stress trials using four levels of clothing (woven clothing, particle barrier, water barrier, and vapor barrier), three levels of relative humidity, and three levels of metabolic rate. Each trial had a compensable and an uncompensable observation plus and observation at the transition point from compensable to uncompensable. Each observation was classified as a case (steady increase in rectal temperature) or non-case (steady rectal temperature). The data were used to compare the difference between the observed metabolic rate (Mobs) and the limiting metabolic rate of TWL (i.e., ∆LimitTWL = Mobs - TWL), where ∆LimitTWL > 0 was above the TWL limit. The sensitivity and specificity for each of the four clothing ensembles were about 0.96 and about 0.20, respectively. Logistic regression for all the data found that ∆LimitTWL, clothing, metabolic rate, and water vapor pressure were significant predictors of outcome. The ln(odds) equations for each clothing ensemble predicted a probability of an uncompensable exposure. The probability of an uncompensable outcome (case) when ∆LimitTWL = 0 was 0.14 for work clothes and particle barrier, and 0.22 for water barrier and vapor barrier. The probability of a case at ∆LimitTWL = 0 was greater than the probability of a case for the wet bulb globe temperature-based exposure limits where the probability of a case was 0.01. That is, the TWL was less restrictive than WBGT but with higher risk.


Estrogen and body weight regulation in women: the role of estrogen receptor alpha (ER-α) on adipocyte lipolysis.
Estrogen has an important role in regulation of fat metabolism. Recent studies indicated that this process occurred due to inhibition of lipolysis by external estrogen administration. However, there was limited information regarding molecular process responsible for this phenomenon. This paper was aimed to present a brief update on recent studies explaining the effect of estrogen on adipose tissue lipolysis and the molecular pathway involved in this process. It is suggested that the effect of estrogen to reduction of lipolysis was through activation of estrogen receptor alpha (ER-α) in adipose tissue. This finding is supported by the fact that mice lacking of ESR1 gene (encodes ER-α) accumulate more fat and ESR1 mRNA in human adipose tissue was inversely correlated with body mass index (BMI). Future study should be aimed to clarify the role of ER-α on lipolysis in adipose tissue during weight loss intervention. Additionally, new pharmacological or nutritional treatment with ability to modulate ER-α activity/expression could be used as a potential weight loss intervention.


Cooperative Hunting Harris' Hawks (Parabuteo unicinctus).
Coordinated hunting by several individuals directed toward the capture and sharing of one Large prey animal has been documented convincingly only for a few mammalian carnivores. In New Mexico, Harris' hawks formed hunting parties of two to six individuals in the nonbreeding season. This behavior improved capture success and the average energy available per individual enabled hawks to dispatch prey larger than themselves. These patterns suggest that cooperation is important to understanding the evolution of complex social behavior in higher vertebrates and, specifically, that benefits derived from team hunting a key factor in the social living of Harris' hawks.


Regulation of vascular function by RCAN1 (ADAPT78).
RCAN1 (Adapt78) functions mainly, if not exclusively, as a regulator of calcineurin, a phosphatase that mediates many cellular responses to calcium. Identification of this regulatory activity has led to a surge of interest in RCAN1, since calcineurin is involved in many cellular and tissue functions, and its abnormal expression is associated with multiple pathologies. Recent studies have implicated RCAN1 as a regulator of angiogenesis. To more fully investigate the role of RCAN1 in vascular function, we first extended previous studies by assessing RCAN1 response in cultured endothelial cells to various vascular agonists. Strong induction of isoform 4 but not isoform 1 was observed in human umbilical vein- and bovine pulmonary aortic-endothelial cells in response to VEGF, thrombin, and ATP but not other agonists. Inductions were both calcium and calcineurin dependent, with the relative effect of each agonist cell-type dependent. Ectopic RCAN1 expression also inhibited calcineurin signaling in the HUVEC cells. Based on these strong RCAN1 responses and a lack of RCAN1-associated vascular studies beyond angiogenesis, we investigated the potential role of RCAN1 in vascular tone using whole mounted mesenteric artery. RCAN1 knockout mice exhibited an attenuated mesenteric vasoconstriction to phenylephrine as compared with wild-type. Overall contractility was unaffected, suggesting that this component of smooth muscle action is similar in the two mouse strains. Constriction in the knockout artery appeared to be potentiated by the addition of the nitric oxide synthase (NOS) inhibitor l-NAME, suggesting that elevated nitric oxide (NO) production occurs in the knockout vasculature and contributes to the weakened vasoconstriction. Our results reveal a newly identified vascular role for RCAN1, and a potential new target for treating vascular- and calcineurin-related disorders.


Multi-disciplinary management for patients with oligometastases to the brain: results of a 5 year cohort study.
The incidence of oligometastases to the brain in good performance status patients is increasing due to improvements in systemic therapy and MRI screening, but specific management pathways are often lacking. We established a multi-disciplinary brain metastases clinic with specific referral guidelines and standard follow-up for good prognosis patients with the view that improving the process of care may improve outcomes. We evaluated patient demographic and outcome data for patients first seen between February 2007 and November 2011. The clinic was feasible to run and referrals were appropriate. 87% of patients referred received a localised therapy during their treatment course. 114 patients were seen and patient numbers increased during the 5 years that the clinic has been running as relationships between clinicians were developed. Median follow-up for those still alive was 23.1 months (6.1-79.1 months). Primary treatments were: surgery alone 52%, surgery plus whole brain radiotherapy (WBRT) 9%, radiosurgery 14%, WBRT alone 23%, supportive care 2%. 43% received subsequent treatment for brain metastases. 25%, 11% and 15% respectively developed local neurological progression only, new brain metastases only or both. Median overall survival following brain metastases diagnosis was 16.0 months (range 1-79.1 months). Breast (32%) and NSCLC (26%) were the most common primary tumours with median survivals of 26 and 16.9 months respectively (HR 0.6, p=0.07). Overall one year survival was 55% and two year survival 31.5%. 85 patients died of whom 37 (44%) had a neurological death. Careful patient selection and multi-disciplinary management identifies a subset of patients with oligometastatic brain disease who benefit from aggressive local treatment. A dedicated joint neurosurgical/ neuro-oncology clinic for such patients is feasible and effective. It also offers the opportunity to better define management strategies and further research in this field. Consideration should be given to defining specific management pathways for these patients within general oncology practice.


Enhanced Surgical Decision-Making Tools in Breast Cancer: Predicting 2-Year Postoperative Physical, Sexual, and Psychosocial Well-Being following Mastectomy and Breast Reconstruction (INSPiRED 004).
We sought to predict clinically meaningful changes in physical, sexual, and psychosocial well-being for women undergoing cancer-related mastectomy and breast reconstruction 2 years after surgery using machine learning (ML) algorithms trained on clinical and patient-reported outcomes data. We used data from women undergoing mastectomy and reconstruction at 11 study sites in North America to develop three distinct ML models. We used data of ten sites to predict clinically meaningful improvement or worsening by comparing pre-surgical scores with 2 year follow-up data measured by validated Breast-Q domains. We employed ten-fold cross-validation to train and test the algorithms, and then externally validated them using the 11th site's data. We considered area-under-the-receiver-operating-characteristics-curve (AUC) as the primary metric to evaluate performance. Overall, between 1454 and 1538 patients completed 2 year follow-up with data for physical, sexual, and psychosocial well-being. In the hold-out validation set, our ML algorithms were able to predict clinically significant changes in physical well-being (chest and upper body) (worsened: AUC range 0.69-0.70; improved: AUC range 0.81-0.82), sexual well-being (worsened: AUC range 0.76-0.77; improved: AUC range 0.74-0.76), and psychosocial well-being (worsened: AUC range 0.64-0.66; improved: AUC range 0.66-0.66). Baseline patient-reported outcome (PRO) variables showed the largest influence on model predictions. Machine learning can predict long-term individual PROs of patients undergoing postmastectomy breast reconstruction with acceptable accuracy. This may better help patients and clinicians make informed decisions regarding expected long-term effect of treatment, facilitate patient-centered care, and ultimately improve postoperative health-related quality of life.


Density embedded VB/MM: a hybrid ab initio VB/MM with electrostatic embedding.
A hybrid QM/MM method that combines ab initio valence-bond (VB) with molecular mechanics (MM) is presented. The method utilizes the ab initio VB approach to describe the reactive fragments and MM to describe the environment thus allows VB calculations of reactions in large biological systems. The method, termed density embedded VB/MM (DE-VB/MM), is an extension of the recently developed VB/MM method. It involves calculation of the electrostatic interaction between the reactive fragments and their environment using the electrostatic embedding scheme. Namely, the electrostatic interactions are represented as one-electron integrals in the ab initio VB Hamiltonian, hence taking into account the wave function polarization of the reactive fragments due to the environment. Moreover, the assumptions that were utilized in an earlier version of the method, VB/MM, to formulate the electrostatic interactions effect on the off-diagonal matrix elements are no longer required in the DE-VB/MM methodology. Using DE-VB/MM, one can calculate, in addition to the adiabatic ground state reaction profile, the energy of the diabatic VB configurations as well as the VB state correlation diagram for the reaction. The abilities of the method are exemplified on the identity SN2 reaction of a chloride anion with methyl chloride in aqueous solution. Both the VB configurations diagram and the state correlation diagram are presented. The results are shown to be in very good agreement with both experimental and other computational data, suggesting that DE-VB/MM is a proper method for application to different reactivity problems in biological systems.


Molecular characteristics of chicken liver arginase.
A purification procedure for the preparation of chicken liver arginase in a homogeneous form is presented. The enzyme hydrolyses both arginine and argininic acid. Kinetic analysis reveals that the enzyme binds arginine when the amino group is protonated or unprotonated; however, the unprotonated form seems to be hydrolysed more rapidly. The enzyme exchanges with the medium approx. 1.6Mn(2+) ions per molecule.


PTEN loss promotes Warburg effect and prostate cancer cell growth by inducing FBP1 degradation.
Fructose-1,6-bisphosphatase (FBP1) is a tumor suppressor and a key enzyme negatively regulating Warburg effect in cancer. However, regulation of FBP1 protein expression and its exact role in prostate cancer (PCa) is largely unclear. Phosphatase and tensin homolog (PTEN) is one of the most frequently deleted tumor suppressor genes in human PCa. However, the role of PTEN loss in aberrant Warburg effect in cancer remains poorly understood. Expression of PTEN and FBP1 was analyzed in several PCa cell lines and prostate tumor tissues in mice. Western blot (WB) and RT-PCR approaches were used to examine how PTEN regulates FBP1 expression. Co-immunoprecipitation (co-IP) and in vivo ubiquitination assays were used to define the regulatory mechanisms. A PCa xenograft model was employed to determine the impact of PTEN regulation of FBP1 on PCa growth in vivo. We demonstrated that in a manner dependent of PI3K/AKT signal pathway PTEN regulated FBP1 expression in various PCa cell lines and tumors in mice. We confirmed that this regulation took place at the protein level and was mediated by SKP2 E3 ubiquitin ligase. Mechanistically, we showed that serine 271 phosphorylation of FBP1 by cyclin-dependent kinases (CDKs) was essential for SKP2-mediated degradation of FBP1 protein induced by PTEN loss. Most importantly, we further showed that loss of PTEN expression enhanced Warburg effect and PCa growth in mice in a manner dependent, at least partially on FBP1 protein degradation. Our results reveal a novel tumor-suppressive feature of PTEN in restraining FBP1 degradation and the Warburg effect. These results also suggest that prohibiting FBP1 protein degradation could be a viable therapeutic strategy for PTEN-deficient PCa.


Congenital malaria by Plasmodium falciparum.
Introdution: Congenital malaria (CM) is a Plasmodium spp infection acquired in utero or during delivery with nonspecific clinical manifestations. Plasmodium falciparum can cause severe illness in pregnant wo men and newborns. to describe two cases of CM caused by Plasmodium falciparum, di fferential diagnosis of sepsis in newborns of pregnant women who live in or have visited endemic malaria zones. Female neonates born in a non-endemic malaria area, diagnosed with neonatal sepsis and treated with antibiotics without clinical response. After the first week of life, the peripheral blood smear identified trophozoites of Plasmodium falciparum thus the newborns were treated with intravenous quinine, improving their condition. The mothers of the two newborns who had malaria in pregnancy, one of them received treatment and she was asymptomatic, and the other one had severe malaria at the time of delivery. CM can cause severe neonatal disease with non-specific, sepsis-like clinical manifestations in which early treatment decreases the risk of complicated malaria. It is a differential diagnosis in newborns of women with a history of malaria during pregnancy or pregnant women visiting or living in endemic malaria areas.


Transversions have larger regulatory effects than transitions.
Transversions (Tv's) are more likely to alter the amino acid sequence of proteins than transitions (Ts's), and local deviations in the Ts:Tv ratio are indicative of evolutionary selection on genes. Whether the two different types of mutations have different effects in non-protein-coding sequences remains unknown. Genetic variants primarily impact gene expression by disrupting the binding of transcription factors (TFs) and other DNA-binding proteins. Because Tv's cause larger changes in the shape of a DNA backbone, we hypothesized that Tv's would have larger impacts on TF binding and gene expression. Here, we provide multiple lines of evidence demonstrating that Tv's have larger impacts on regulatory DNA including analyses of TF binding motifs and allele-specific TF binding. In these analyses, we observed a depletion of Tv's within TF binding motifs and TF binding sites. Using massively parallel population-scale reporter assays, we also provided empirical evidence that Tv's have larger effects than Ts's on the activity of human gene regulatory elements. Tv's are more likely to disrupt TF binding, resulting in larger changes in gene expression. Although the observed differences are small, these findings represent a novel, fundamental property of regulatory variation. Understanding the features of functional non-coding variation could be valuable for revealing the genetic underpinnings of complex traits and diseases in future studies.


Systemic blockade of P2X3 and P2X2/3 receptors attenuates bone cancer pain behaviour in rats.
Pain remains an area of considerable unmet clinical need, and this is particularly true of pain associated with bone metastases, in part because existing analgesic drugs show only limited efficacy in many patients and in part because of the adverse side effects associated with these agents. An important issue is that the nature and roles of the algogens produced in bone that drive pain-signalling systems remain unknown. Here, we tested the hypothesis that adenosine triphosphate is one such key mediator through actions on P2X3 and P2X2/3 receptors, which are expressed selectively on primary afferent nocioceptors, including those innervating the bone. Using a well-established rat model of bone cancer pain, AF-353, a recently described potent and selective P2X3 and P2X2/3 receptor antagonist, was administered orally to rats and found to produce highly significant prevention and reversal of bone cancer pain behaviour. This attenuation occurred without apparent modification of the disease, since bone destruction induced by rat MRMT-1 carcinoma cells was not significantly altered by AF-353. Using in vivo electrophysiology, evidence for a central site of action was provided by dose-dependent reductions in electrical, mechanical and thermal stimuli-evoked dorsal horn neuronal hyperexcitability following direct AF-353 administration onto the spinal cord of bone cancer animals. A peripheral site of action was also suggested by studies on the extracellular release of adenosine triphosphate from MRMT-1 carcinoma cells. Moreover, elevated phosphorylated-extracellular signal-regulated kinase expression in dorsal root ganglion neurons, induced by co-cultured MRMT-1 carcinoma cells, was significantly reduced in the presence of AF-353. These data suggest that blockade of P2X3 and P2X2/3 receptors on both the peripheral and central terminals of nocioceptors contributes to analgesic efficacy in a model of bone cancer pain. Thus, systemic P2X3 and P2X2/3 receptor antagonists with central nervous system penetration may offer a promising therapeutic tool in treating bone cancer pain.


Perceptions about the acceptability and prevalence of HIV testing and factors influencing them in different communities in South Africa.
HIV counselling and testing (HCT) is considered important because it is an entry point to a comprehensive continuum of care for HIV/AIDS. The South African Department of Health launched an HCT campaign in April 2010, and this reached 13,269,746 people by June 2011, of which 16% tested HIV positive and 400,000 of those were initiated into antiretroviral treatment. The overall objective of this project was to gain insight into the general perceptions about HIV testing in the different South African communities. Factors influencing testing in these communities were also explored. Discussions with twelve focus groups (FG) of 8-12 participants each were conducted with male and female participants recruited from both urban formal and informal communities in Cape Town and Durban. Participants included four racial groups represented by different age groups as follows: adolescents (12-17 years), youth (18-24 years) and adults (25 years and older). Data were analyzed using thematic coding. Among the key themes that emerged from the findings were the inaccurate perception of risk, fear of testing HIV positive, stigma and discrimination. Participants from both African and Indian FGs reported being less likely to do self-initiated HIV testing and counselling, while those from the FG consisting of young whites were more likely to learn about their HIV status through blood donations and campus HIV testing campaigns. Most FGs said they were likely to test if they understood the testing process better and also if the results are kept confidential. The present findings reiterate the importance of spreading positive messages and ensuring confidentiality for HIV testing in a society where there is still some stigma associated with people living with HIV/AIDS. This can partly be accomplished by the continuation of the national HCT campaign, which has been a considerable success in the fight against HIV/AIDS in South Africa during the past two years.


Transcription factor ISL1 is essential for pacemaker development and function.
The sinoatrial node (SAN) maintains a rhythmic heartbeat; therefore, a better understanding of factors that drive SAN development and function is crucial to generation of potential therapies, such as biological pacemakers, for sinus arrhythmias. Here, we determined that the LIM homeodomain transcription factor ISL1 plays a key role in survival, proliferation, and function of pacemaker cells throughout development. Analysis of several Isl1 mutant mouse lines, including animals harboring an SAN-specific Isl1 deletion, revealed that ISL1 within SAN is a requirement for early embryonic viability. RNA-sequencing (RNA-seq) analyses of FACS-purified cells from ISL1-deficient SANs revealed that a number of genes critical for SAN function, including those encoding transcription factors and ion channels, were downstream of ISL1. Chromatin immunoprecipitation assays performed with anti-ISL1 antibodies and chromatin extracts from FACS-purified SAN cells demonstrated that ISL1 directly binds genomic regions within several genes required for normal pacemaker function, including subunits of the L-type calcium channel, Ank2, and Tbx3. Other genes implicated in abnormal heart rhythm in humans were also direct ISL1 targets. Together, our results demonstrate that ISL1 regulates approximately one-third of SAN-specific genes, indicate that a combination of ISL1 and other SAN transcription factors could be utilized to generate pacemaker cells, and suggest ISL1 mutations may underlie sick sinus syndrome.


Burden of illness of bacterial cellulitis and erysipelas of the leg in the Netherlands.
Information on the prevalence of bacterial cellulitis (BC) and erysipelas (ER) of the leg (BCERL) is sparse and dependent on the definitions used. There is no information available on the number of hospitalized and non-hospitalized patients with BCERL, and related treatment costs. The purpose of this study was to assess the burden of illness for BCERL in the Netherlands in 2001. Data were obtained from different linked databases. Hospital information was obtained from the National Morbidity Registration (known in the Netherlands as the LMR), which includes all Dutch citizens, using ICD-9-CM codes. The number of patients not admitted to hospital was estimated using a subsample with data from general practitioners (GPs) (N = 50,000). These data were extrapolated using age/gender and disease-specific standardization. The subsample was used to assess the location of the infection. Reimbursement costs were available for all resources. In 2001, approximately 28,000 patients presented with either BC or ER of the leg. Of these patients, 2,200 were admitted to the hospital and 4-6% had two or more episodes of ER/BC in 2001. The average costs per hospitalization for BCERL were 5,346 euros, accumulating to more than 14 million euros in 2001. Although only 7% of all patients were hospitalized, 83% of the total treatment costs could be attributed to hospitalization. BCERL are common and serious infections in the Netherlands. Hospitalization occurs in only one in 14 patients but contributes more than 80% of the total costs, which accumulate to 17 million euros a year.


Where we stand on proficiency testing.
Because of the lack of comprehensive information in radiologic technology publications on the history, purpose and future implications of a process technologists hear talked about as "proficiency testing," and because of a lack of understanding of the role The American Society of Radiologic Technologists is playing in the project, the authors responded to a request by the editor to prepare this article. Since evaluation of the proficiency testing concept is still going on, this article is only a beginning and cannot be conclusive as to possible implementation, but it serves to provide substantial understanding through which the reader can evaluate future information.


AMP- and stress-activated protein kinases: key regulators of glucose-dependent gene transcription in mammalian cells?
This article will discuss the role of two classes of serine/threonine protein kinases in the regulation of gene transcription in mammals. The first is AMP-activated protein kinase (AMPK), which is responsive to changes in the intracellular energy status. The second is the 'stress-activated" family of protein kinases, members of the mitogen-activated protein (MAP) kinase superfamily, whose regulation by a number of extracellular agents (including osmotic stresses, cytokines, and heat) is less well understood. Interest in these enzymes has grown in the past few years due to mounting evidence (both pharmacological and genetic) which has implicated them in the regulation of a number genes important in mammalian metabolism.


The tumor necrosis factor family receptors RANK and CD40 cooperatively establish the thymic medullary microenvironment and self-tolerance.
Medullary thymic epithelial cells (mTECs) establish T cell self-tolerance through the expression of autoimmune regulator (Aire) and peripheral tissue-specific self-antigens. However, signals underlying mTEC development remain largely unclear. Here, we demonstrate crucial regulation of mTEC development by receptor activator of NF-kappaB (RANK) and CD40 signals. Whereas only RANK signaling was essential for mTEC development during embryogenesis, in postnatal mice, cooperation between CD40 and RANK signals was required for mTEC development to successfully establish the medullary microenvironment. Ligation of RANK or CD40 on fetal thymic stroma in vitro induced mTEC development in a tumor necrosis factor-associated factor 6 (TRAF6)-, NF-kappaB inducing kinase (NIK)-, and IkappaB kinase beta (IKKbeta)-dependent manner. These results show that developmental-stage-dependent cooperation between RANK and CD40 promotes mTEC development, thereby establishing self-tolerance.


Towards better understanding of the photophysics of platinum(II) coordination compounds with anthracene- and pyrene-substituted 2,6-bis(thiazol-2-yl)pyridines.
The photophysical properties of platinum(ii) compounds with 4-(9-anthryl)-2,6-bis(thiazol-2-yl)pyridine (1) and 4-(1-pyrenyl)-2,6-bis(thiazol-2-yl)pyridine (2) were widely investigated. In DMSO and MeCN, the triplet emission of (1) and (2) most probably experiences the solvent-induced exciplex quenching and both complexes exhibit ligand-based fluorescence with maxima in the range of 468-570 nm. In non-coordinating dichloromethane, the emission of (1) and (2) shifts to longer wavelengths, and the lifetimes in microseconds are indicative of phosphorescence. The formation of the triplet excited state was further supported by the observation of singlet oxygen photoluminescence in the near-IR at 1270 nm. To explore the nature of the triplet excited state, the emission properties of (1) and (2) in low temperature glasses were analysed in comparison with those for free ligands and appropriate hydrocarbons, as well as transient absorption spectra were recorded in femtosecond and nanosecond regimes. The studies revealed that the lowest energy triplet state of both complexes is an admixture of 3ILCT and 3(π-π*)aryl character, but the contribution of the 3ILCT state in (1) originating from anthracene to 2,6-bis(thiazol-2-yl)pyridine can be determined as rather negligible.


Ascorbate deficiency increases progression of shigellosis in guinea pigs and mice infection models.
Shigella spp. are the causative agents of bacterial dysentery and shigellosis, mainly in children living in developing countries. The study of Shigella entire life cycle in vivo and the evaluation of vaccine candidates' protective efficacy have been hampered by the lack of a suitable animal model of infection. None of the studies evaluated so far (rabbit, guinea pig, mouse) allowed the recapitulation of full shigellosis symptoms upon Shigella oral challenge. Historical reports have suggested that dysentery and scurvy are both metabolic diseases associated with ascorbate deficiency. Mammals, which are susceptible to Shigella infection (humans, non-human primates and guinea pigs) are among the few species unable to synthesize ascorbate. We optimized a low-ascorbate diet to induce moderate ascorbate deficiency, but not scurvy, in guinea pigs to investigate whether poor vitamin C status increases the progression of shigellosis. Moderate ascorbate deficiency increased shigellosis symptom severity during an extended period of time (up to 48 h) in all strains tested (Shigella sonnei, Shigella flexneri 5a, and 2a). At late time points, an important influx of neutrophils was observed both within the disrupted colonic mucosa and in the luminal compartment, although Shigella was able to disseminate deep into the organ to reach the sub-mucosal layer and the bloodstream. Moreover, we found that ascorbate deficiency also increased Shigella penetration into the colon epithelium layer in a Gulo-/- mouse infection model. The use of these new rodent models of shigellosis opens new doors for the study of both Shigella infection strategies and immune responses to Shigella infection.


Evidence for a nonallelic heterogeneity of epidermodysplasia verruciformis with two susceptibility loci mapped to chromosome regions 2p21-p24 and 17q25.
Epidermodysplasia verruciformis is a rare genodermatosis associated with a high risk of skin cancer. This condition is characterized by an abnormal susceptibility to specific related human papillomavirus genotypes, including the oncogenic HPV5. Epidermodysplasia verruciformis is usually considered as an autosomal recessive disease. We recently mapped a susceptibility locus for epidermodysplasia verruciformis (EV1) to chromosome 17qter within the 1 cM interval between markers D17S939 and D17S802. We report here the genotyping for 10 microsatellite markers spanning 29 cM around EV1 in two consanguineous epidermodysplasia verruciformis families from Colombia (C2) and France (F1) comprising five patients and two patients, respectively. Using homozygosity mapping, linkage with 17qter markers was observed for family C2 only. Multipoint linkage analysis yielded maximum multipoint LOD-score values above 10 between markers D17S1839 and D17S802 encompassing the EV1 locus. A genome-wide search performed in family F1 yielded evidence for linkage between epidermodysplasia verruciformis and the chromosomal 2p marker D2S365. Nine additional microsatellite markers spanning 15 cM in this region were analyzed. Assuming an autosomal recessive inheritance with a complete penetrance, the expected maximum two-point LOD-score value of 1.8 was obtained for three markers and multipoint linkage analysis yielded a maximum LOD-score value of 3. 51 between markers D2S2144 and D2S392. Haplotype analysis allowed to map a candidate region for a second epidermodysplasia verruciformis susceptibility locus (EV2) within the 8 cM interval between markers D2S171 and D2S2347 of the 2p21-p24 region. In contrast, linkage with 2p markers was excluded for family C2 and for the three families in which we mapped EV1 previously. The disclosure of two susceptibility loci for epidermodysplasia verruciformis provides evidence for a nonallelic heterogeneity in this disease.


Sub-THz Vibrational Dynamics in Ordered Mesoporous Silica Nanoparticles.
The vibrational dynamics in the sub-THz range of mesoporous silica nanoparticles (MSNs) having ordered cylindrical mesopores was investigated. MCM-41 and SBA-15 particles were synthesized, and their structure was determined using scanning electron microscopy (SEM), low-angle X-ray diffraction (XRD), N2 physisorption analyses, and Raman scattering. Brillouin scattering measurements are reported and enabled determining the stiffness of the silica walls (speed of sound) using finite element calculations for the ordered mesoporous structure. The relevance of this approach is discussed based on the comparison between the numerical and experimental results and previous works reported in the literature.


Recombinant Rhodobacter capsulatus xanthine dehydrogenase, a useful model system for the characterization of protein variants leading to xanthinuria I in humans.
Rhodobacter capsulatus xanthine dehydrogenase (XDH) forms an (alphabeta)2 heterotetramer and is highly homologous to homodimeric eukaryotic XDHs. The crystal structures of bovine XDH and R. capsulatus XDH showed that the two proteins have highly similar folds. We have developed an efficient system for the recombinant expression of R. capsulatus XDH in Escherichia coli. The recombinant protein shows spectral features and a range of substrate specificities similar to bovine milk xanthine oxidase. However, R. capsulatus XDH is at least 5 times more active than bovine XDH and, unlike mammalian XDH, does not undergo the conversion to the oxidase form. EPR spectra were obtained for the FeS centers of the enzyme showing an axial signal for FeSI, which is different from that reported for xanthine oxidase. X-ray absorption spectroscopy at the iron and molybdenum K-edge and the tungsten LIII-edge have been used to probe the different metal coordinations of variant forms of the enzyme. Based on a mutation identified in a patient suffering from xanthinuria I, the corresponding arginine 135 was substituted to a cysteine in R. capsulatus XDH, and the protein variant was purified and characterized. Two different forms of XDH-R135C were purified, an active (alphabeta)2 heterotetrameric form and an inactive (alphabeta) heterodimeric form. The active form contains a full complement of redox centers, whereas in the inactive form the FeSI center is likely to be missing.


Genetic content and preliminary transcriptional analysis of a representative region of murine gammaherpesvirus 68.
Murine gammaherpesvirus 68 (MHV-68) is a relatively recently discovered pathogen of wild rodents and provides a unique opportunity to explore in detail the interactions of a gammaherpesvirus with its natural host. It may also provide a much needed small animal model for human gammaherpesviruses. As a step in the detailed analysis of virus gene structure and expression we have sequenced over 20 kb of the MHV-68 genome and mapped gene transcripts by Northern blot hybridization. The region we chose to analyse contains several conserved gene blocks as well as some less well conserved genes and allowed us to estimate the relationship of this virus to other herpesvirus family members. Of particular interest is the fact that none of the characteristic Epstein-Barr virus (EBV) genes is present at this genomic locus although MHV-68 does have one gene encoding a membrane glycoprotein, 9p150, which shows similarities to the major membrane glycoprotein of EBV. Our results further confirm that MHV-68 is a gammaherpesvirus marginally more closely related to a cluster of gammaherpesviruses including herpesvirus salmiri than to EBV. Northern analysis shows that the temporal regulation of expression is broadly similar to that of other herpesviruses in this region of the genome. We also show that like other gammaherpesviruses, MHV-68 splices its homologue of the EBV transcriptional activator gene BMRF1.


Advances in improving fertility in women through stem cell-based clinical platforms.
Due to their regenerative ability, stem cells are looked at as a promising tool for improving infertility treatments in women. As the main limiting factor in female fertility is represented by the decrease of ovarian reserve, the main goals of stem cell-based clinical platforms would be to obtain in vitro or in vivo neo-oogenesis. Refractory endometrial factor infertility also represents an obstacle for female reproduction for which stem cells might provide novel treatment strategies. Areas covered: A systematic search of the literature was performed on MEDLINE/PubMed database to identify relevant articles using stem-cell based clinical or research platforms in the field of female infertility. Expert opinion: In vitro oogenesis has not so far developed beyond the stage of oocyte-like cells whose normal progression to mature oocytes and ability to be fertilized was not proved. Extensive epigenetic programming of gamete precursors and the complex interactions between somatic and germ cells required for human oogenesis likely represent the main obstacles in stem-cell-based neo-oogenesis. Also resuming oogenesis in vivo in adulthood still appears a distant hypothesis, as there is still a lack of consensus about the existence and functionality of adult ovarian stem cells.


Regiospecific synthesis of tetrasubstituted phthalocyanines and their liquid crystalline order.
Metal-free and metal(II) all-endo-tetraalkoxy-phthalocyanines of C4h symmetry are synthesised regiospecifically from 3-(2-butyloctyloxy)phthalonitrile with lithium octanolate and subsequent metal ion exchange. The voluminous, yet not overly large, and racemically disordered alkoxy substituent not only renders the cyclotetramerisation regiospecific, but also leads to liquid crystalline self-assembly with attainable clearing temperatures and persisting columnar organisation at room temperature. A rare hexagonal mesophase with twelve columns per hexagonal unit cell is found in the metal-free homologue, whereas the metal complexes show rectangular mesophases. The clearing temperature increases with increasing axial component of the metal ion coordination sphere. At low temperature, significant antiferromagnetic exchange between magnetic centres is observed for the Co(II) homologue, whereas the magnetic centres are magnetically independent in the Cu(II) derivative, in line with the observed higher clearing temperature in the Co(II) case that testifies of stronger interdisk interactions.


Computational approaches to identify functional genetic variants in cancer genomes.
The International Cancer Genome Consortium (ICGC) aims to catalog genomic abnormalities in tumors from 50 different cancer types. Genome sequencing reveals hundreds to thousands of somatic mutations in each tumor but only a minority of these drive tumor progression. We present the result of discussions within the ICGC on how to address the challenge of identifying mutations that contribute to oncogenesis, tumor maintenance or response to therapy, and recommend computational techniques to annotate somatic variants and predict their impact on cancer phenotype.


Embedding Evidence-based Practice Education into a Post-graduate Physiotherapy Program: Eight Years of pre-Post Course Evaluations.
Little has been published about the effectiveness of training postgraduate physiotherapy coursework students in research methods and evidence-based practice (EBP) theory. Graduate qualities in most universities include lifelong learning. Inclusion of EBP in post-graduate coursework students' training is one way for students to develop the knowledge and skills needed to implement current best evidence in their clinical practice after graduation, thereby facilitating lifelong learning. This paper reports on change in confidence and anxiety in knowledge of statistical terminology and concepts related to research design and EBP in eight consecutive years of post-graduate physiotherapy students at one Australian university. Pre-survey/post-survey instruments were administered to students in an intensive 3-week post-graduate course, which taught health research methods, biostatistics and EBP. This course was embedded into a post-graduate physiotherapy programme from 2007 to 2014. The organization and delivery of the course was based on best pedagogical evidence for effectively teaching adult physiotherapists. The course was first delivered each year in the programme, and no other course was delivered concurrently. There were significant improvements in confidence, significantly decreased anxiety and improvements in knowledge of statistical terminology and concepts related to research design and EBP, at course completion. Age, gender and country of origin were not confounders on learning outcomes, although there was a (non-significant) trend that years of practice negatively impacted on learning outcomes (p = 0.09). There was a greater improvement in confidence in statistical terminology than in concepts related to research design and EBP. An intensive teaching programme in health research methods and biostatistics and EBP, based on best practice adult physiotherapy learning principles, is effective immediately post-course, in decreasing anxiety and increasing confidence in the terminology used in research methods and EBP. Copyright © 2016 John Wiley & Sons, Ltd.


A review of the pharmacological mechanism of traditional Chinese medicine in the intervention of coronary heart disease and stroke.
In recent years, several researches have showed that Buyang Huanwu Decoction (BHD) possesses multiple target points in the intervention of diseases, and has same treatment effects on cerebrovascular diseases and cardiovascular diseases. But, there was no full report about the mechanistic and material basis in Brain-Heart concurrent treatment. The objective of the present study was to examine the pharmacological mechanism of traditional Chinese medicine in the intervention of coronary heart disease and stroke. We combined the HIT, PubChem, David Database resource and the networked pharmacology method to ultimately find out BHD's thirty-five potential brain-heart concurrent treatment target points, and preliminarily reveal BHD's material basis for treatment of cerebrovascular diseases and cardiovascular diseases. Finally, the study provided new information with the guidance meanings.


Transforming growth factor {beta}1 induces epithelial-mesenchymal transition by activating the JNK-Smad3 pathway in rat peritoneal mesothelial cells.
Peritoneal fibrosis is a serious complication in long-term peritoneal dialysis (PD) patients. Epithelial-mesenchymal transition (EMT) plays an important role in peritoneal fibrosis, and TGFbeta1 is the crucial inducer of EMT. Phosphorylation of Smad proteins is required for TGFbeta1-induced EMT. It was reported that C-Jun N-terminal kinase (JNK) was involved in the TGFbeta1/Smad signaling pathway and might regulate the activation of Smad proteins. However, whether JNK is activated by TGFbeta1 in rat peritoneal mesothelial cells (RPMCs) and the role taken by JNK signaling in EMT induced by TGFbeta1 remains undetermined. In the present study, we investigated the role of JNK-Smad pathway in EMT induced by TGFbeta1 in RPMCs. We harvested RPMCs from the peritoneum of male Sprague-Dawley rats and then cultured the cells in Dulbecco modified Eagle medium/F12 medium with 15% (volume:volume) fetal bovine serum. The cells were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGFbeta1. The protein expression levels of phosphorylated JNK, Smad2, and Smad3 were detected by Western blotting. The messenger RNA levels and protein expression of alpha-smooth muscle actin (alpha-SMA), E-cadherin, and collagen I were determined with reverse transcriptase polymerase chain reaction and Western blotting respectively. Expression of alpha-SMA and collagen I were significantly increased and expression of E-cadherin decreased with TGFbeta1 in RPMCs. Transforming growth factor beta1 can stimulate phosphorylated JNK expression from 5 minutes, with the peak at 10 minutes, and phosphorylated Smad2 and Smad3 expression from 10 minutes, with the peak at 30 minutes. The addition of SP600125, which blocked activation of JNK, effectively inhibited TGFbeta1-induced phosphorylation of Smad3, but not Smad2. Also, our results showed that SP600125 effectively suppressed TGFbeta1-induced high expression of alpha-SMA and collagen I, and prevented TGFbeta1-induced downregulation of E-cadherin expression in RPMCs. This study demonstrated that JNK signaling may play an important role in EMT induced by TGFbeta1 in RPMCs through activation of Smad3, suggesting that JNK inhibitor may prove to be a novel therapeutic agent for peritoneal fibrosis.


Lipoxygenation of arachidonic acid: a pivotal step in stimulus-secretion coupling in the pancreatic beta cell.
The relationship(s) between calcium flux, phospholipid turnover and stimulus-secretion coupling in endocrine cells remains unclear. Several stimuli of insulin secretion promote calcium accumulation in the pancreatic beta cell; recent studies are compatible with the formulation that, as a consequence of such ion flux, arachidonic acid is released from phospholipids by calcium-dependent phospholipase(s), and may, in turn, be metabolized via lipoxygenase to products which promote insulin release. In the current study, we examined the effects of two known lipoxygenase inhibitors, BW755c and nordihydroguaiaretic acid, on the augmentation of insulin release evoked by three pharmacologically distinct classes of insulin secretagogues--one which augments cyclic AMP accumulation (exemplified by theophylline), one which inhibits prostaglandin synthesis (sodium salicylate), and one which is independent of changes in cyclic AMP or prostaglandin availability (tolbutamide). Both inhibitors of lipoxygenase markedly blunted the insulinogenic response to glucose, as well as to all three drugs, without evidence of cell toxicity. These data are most compatible with a central role for lipoxygenase products in coupling insulin secretion to stimuli of widely varying natures.


Stainless steel manual metal arc welding fumes in rats.
Forty two male Wistar rats were exposed to manual metal arc (MMA) stainless steel (SS) welding fumes generated by an automatic welding device for "nose-only" exposure. The exposure simulated an actual MMA/SS welding environment as closely as possible. For the retention study, the duration of exposure was one hour per workday for one, two, three, of four weeks and for the clearance study four weeks. The retention and clearance of the chromium, nickel, and iron found in MMA/SS welding fumes in the rats' lungs were studied as was the distribution of the metals to other organs. Instrumental neutron activation analysis (INAA) was used for the multi-element chemical activation analyses. The concentrations of chromium and nickel in the blood and the urine were determined by atomic absorption method (AAS). The retention of exogenous iron was determined by a magnetic measuring method. The results indicated that the lungs were the target organs of soluble hexavalent chromates. The half times of lung clearance for Cr, Ni, and Fe were 40 +/- 4 d, 20 +/- d, and 50 +/- 10 d. When the lung clearance curves are compared, the half times of Cr and Fe lung clearance are similar but nickel disappears faster. The distribution and clearance patterns of chromium to other organs differ from those obtained after single intravenous or intratracheal injections of alkaline chromates.


Pseudoxanthomonas helianthi sp. nov., isolated from roots of Jerusalem artichoke (Helianthus tuberosus).
A bacterium designated as strain roo10T was isolated from roots of Jerusalem artichoke (Helianthus tuberosus). Cells were Gram-stain-negative and non-motile rods. The phylogenetic analysis of the 16S rRNA gene indicated that it represented a member of the genus Pseudoxanthomonas, and its close relatives included Pseudoxanthomonas kalamensis JA40T (97.8 % 16S rRNA gene sequence similarity), Pseudoxanthomonas sangjuensis 5GH38-5T (97.7 %) and Pseudoxanthomonas daejeonensis TR6-08T (97.1 %). Growth of roo10T occurred at pH 7-9. The temperature for growth ranged from 20 to 37 °C. Tolerance to NaCl was observed from 0.005 to 5 % (w/v) concentration. Predominant fatty acids were iso-C15 : 0 (23.5 %), iso-C16 : 0 (18.9 %) and anteiso-C15 : 0 (11.5 %). Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidyl-N-methylethanolamine were the major polar lipids. The predominant quinone was ubiquinone 8 (Q-8). The DNA G+C content was 65.7 mol% [from melting temperature (Tm)]. Comparison of phenotypic and chemotaxonomic characteristics indicated that roo10T was distinguishable from its close relatives. Additionally, the DNA-DNA relatedness levels between roo10T and P. kalamensis DSM 18571T (22±0.5 %), P. sangjuensis 5GH38-5T (21±0.2 %) and P. daejeonensis DSM 17801T (3±1 %) were lower than 70 %. These results indicated that roo10T represented a novel species of the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas helianthi sp. nov. is proposed. The type strain is roo10T (=BCC 70700T=NBRC 110414T).


Evolutionary Monte Carlo of QM Properties in Chemical Space: Electrolyte Design.
Optimizing a target function over the space of organic molecules is an important problem appearing in many fields of applied science but also a very difficult one due to the vast number of possible molecular systems. We propose an evolutionary Monte Carlo algorithm for solving such problems which is capable of straightforwardly tuning both exploration and exploitation characteristics of an optimization procedure while retaining favorable properties of genetic algorithms. The method, dubbed MOSAiCS (Metropolis Optimization by Sampling Adaptively in Chemical Space), is tested on problems related to optimizing components of battery electrolytes, namely, minimizing solvation energy in water or maximizing dipole moment while enforcing a lower bound on the HOMO-LUMO gap; optimization was carried out over sets of molecular graphs inspired by QM9 and Electrolyte Genome Project (EGP) data sets. MOSAiCS reliably generated molecular candidates with good target quantity values, which were in most cases better than the ones found in QM9 or EGP. While the optimization results presented in this work sometimes required up to 106 QM calculations and were thus feasible only thanks to computationally efficient ab initio approximations of properties of interest, we discuss possible strategies for accelerating MOSAiCS using machine learning approaches.


An abundant hydrophobin (ABH1) forms hydrophobic rodlet layers in Agaricus bisporus fruiting bodies.
The SDS-insoluble protein fraction of Agaricus bisporus fruiting bodies was solubilized with trifluoroacetic acid. On SDS-PAGE this fraction was found to contain one abundant protein with an apparent M(r) of 16 kDa. The N-terminal amino acid sequence of this protein was determined and RT-PCR used to isolate a cDNA clone which upon sequencing identified the protein as a typical class I hydrophobin (ABH1). The gene (ABH1) was isolated and sequenced, and a second hydrophobin gene (ABH2) was found about 2.5 kbp downstream of ABH1. Purified ABH1 self-assembled at hydrophobic-hydrophilic interfaces, producing the typical rodlet layer known from other hydrophobins. Similar rodlets were observed on the surface of the fruiting body, while immunological localization showed the hydrophobin to be particularly abundant at the outer surface of fruiting bodies, in the veil and in the core tissue of the stipe. Transcripts of ABH1 were found only in fruiting-body hyphae. The ABH1 hydrophobin is probably solely responsible for the hydrophobicity of the fruiting-body surface but may also line air channels within fruiting bodies.


Dietary Hemp Seeds More Effectively Attenuate Disorders in Genetically Obese Rats than Their Lipid Fraction.
Hemp seeds are rich in PUFAs and other bioactives that can attenuate the development of obesity-related disorders; however, the extent to which their lipid fraction is responsible for this effect is unknown. We hypothesized that hemp seed or hemp oil supplementation can attenuate genetically determined disorders and that the former are more effective in doing so. Lean and obese male Zucker rats, aged 8 wk, weighing 174 ± 4.2 g and 223 ± 3.8 g, respectively, were allocated to 4 groups. The lean (LC) and obese controls (OC) were fed a standard diet, whereas the other 2 obese groups were fed a modified diet in which hemp oil (4% diet; O + HO) or hemp seeds (12% diet; O + HS) were included. All diets had the same proportions of protein (18%), fat (8%), and fiber (5%) and a similar carbohydrate proportion (∼52%). Diets fed to O + HO and O + HS had similar fatty acid profiles. After 4 wk, markers of gut and liver function, antioxidant status, and lipid metabolism were measured. The total SCFA concentration in the cecal digesta was lower in OC (64.8 ± 4.21 µmol/g) compared with LC (78.1 ± 2.83 µmol/g) (P ≤ 0.05), whereas it was greater in O + HS (89 ± 4.41 µmol/g) compared with LC, OC, and O + HO (69.7 ± 2.68 µmol/g) (P ≤ 0.05). Plasma total cholesterol was greater in OC (6.20 ± 0.198 mmol/L) and O + HO (5.60 ± 0.084 mmol/L) compared with LC (2.71 ± 0.094 mmol/L) (P ≤ 0.05); in O + HS, the concentration did not differ from the other groups (5.16 ± 0.278 mmol/L). The liver cholesterol concentration was greater in OC (1.79 ± 0.379 mg/g) compared with the other groups (1.28-1.43 mg/g) (P ≤ 0.05). Hepatic expression of peroxisome proliferator-activated receptor γ was lower in OC (11.9 ± 0.93 units) compared with LC (17.3 ± 1.3 units) (P ≤ 0.05), whereas it was greater in O + HS (19.2 ± 1.04 units) compared with OC and O + HO (14.0 ± 1.33 units) (P ≤ 0.05). Dietary hemp seeds more effectively attenuate metabolic disorders in genetically obese rats than the oil extracted from them, which suggests that the lipid fraction is only partly responsible for these effects.


The diagnosis and treatment of dyskeratosis congenita: a review.
Dyskeratosis congenita (DC) is an inherited bone marrow failure (BMF) syndrome characterized by the classic triad of abnormal skin pigmentation, nail dystrophy, and oral leukoplakia. However, patients usually develop BMF and are predisposed to cancer, with increased risk for squamous cell carcinoma and hematolymphoid neoplasms. DC is a disease of defective telomere maintenance and is heterogeneous at the genetic level. It can be inherited in X-linked, autosomal dominant, or autosomal recessive patterns. Mutations in at least ten telomere- and telomerase-associated genes have been described in DC. There are no targeted therapies for DC and patients usually die of BMF due to a deficient renewing capability of hematopoietic stem cells. Allogeneic hematopoietic stem cell transplantation is the only curative treatment for BMF.


Somatosensory evoked potentials and EEG findings in siblings of juvenile myoclonic epilepsy patients.
In order to demonstrate cortical hyperexitability and to determine the efficacy of 'SEP of high amplitude (HASEP) in detecting asymptomatic relatives of JME patients, we evaluated the EEG findings and SEP in asymptomatic siblings of JME patients. We analysed the EEG and SEP findings of 37 JME patients and 48 asymptomatic siblings of 27 patients and compared the results with healthy volunteers. Of 13 asymptomatic siblings with EEG abnormalities (27.1%), 5 had 4-6 Hz spike/polyspike and wave paroxysms (10.4%), 5 had slow wave paroxysms (10.4%), and the remaining 3 patients exhibited focal spike and wave activity during hyperventilation (6.25%). No significant difference was found between JME patients, asymptomatic siblings and control groups with respect to N20 latencies. N20/P25 amplitudes were significantly higher in both JME group and asymptomatic siblings than those of control cases. HASEP were observed in 8 out of 37 JME patients (21.6%), 10 out of 48 healthy siblings (20.8%) and none of the control cases. The consanguinity rate was higher in parents of patients who had at least one asymptomatic sibling with EEG and/or SEP abnormalities (68.7%) than in those of patients who had siblings with no EEG and SEP abnormalities (9.9%). We suggest that the high rate of SEP and EEG changes among offspring of consanguineous marriages might reflect the genetic heterogeneity of the disease.


Widespread distribution and rate differentiation of the atrial pacemaker complex.
In a study to examine the basis of rate-related changes in the electrocardiographic P wave we found a multicentric rather than unifocal origin of the atrial depolarization wave in dogs. Three to five pacemakers, or origin points, were distributed over a 30- to 40-mm area compared to the 11-mm size of the sinus node. Two or three of the sites could excite simultaneously, or one specific site would dominate excitation. Each separate origin point dominated excitation within a specific range of heart rates, and on reaching either the upper or lower limits of this range, a new focus abruptly dominated and initiated the atrial wave front. We have obtained evidence to suggest that these findings may be explained by a widely distributed atrial pacemaker complex. The spatial distribution of this system exceeded the dimensions of the canine sinus node by a factor of three to four times. The pacemaker centers, although distributed, were consistently located at specific positions along the superior vena caval-right atrial junction. Also, each separate pacemaker site appeared functionally differentiated to generate a specific range of heart rates. We propose that in addition to the sinus node there are other specialized atrial pacemaker centers, and that this specialization, including the differentiated response and coordination, is conferred by focal receptor characteristics and their inputs.


The effect of three hemostatic agents on early bone healing in an animal model.
Resorbable bone hemostasis materials, oxidized regenerated cellulose (ORC) and microfibrillar collagen (MFC), remain at the site of application for up to 8 weeks and may impair osteogenesis. Our experimental study compared the effect of a water-soluble alkylene oxide copolymer (AOC) to ORC and MFC versus no hemostatic material on early bone healing. Two circular 2.7 mm non-critical defects were made in each tibia of 12 rabbits. Sufficient AOC, ORC or MFC was applied to achieve hemostasis, and effectiveness recorded. An autologous blood clot was applied to control defects. Rabbits were sacrificed at 17 days, tibiae excised and fixed. Bone healing was quantitatively measured by micro-computed tomography (micro-CT) expressed as fractional bone volume, and qualitatively assessed by histological examination of decalcified sections. Hemostasis was immediate after application of MFC and AOC, after 1-2 minutes with ORC, and >5 minutes for control. At 17 days post-surgery, micro-CT analysis showed near-complete healing in control and AOC groups, partial healing in the ORC group and minimal healing in the MFC group. Fractional bone volume was 8 fold greater in the control and AOC groups than in the MFC group (0.42 ± 0.06, 0.40 ± 0.03 vs 0.05 ± 0.01, P < 0.001) and over 1.5-fold greater than in the ORC group (0.25 ± 0.03, P < 0.05). By histology, MFC remained at the application site with minimal healing at the defect margins and early fibrotic tissue within the defect. ORC-treated defects showed partial healing but with early fibrotic tissue in the marrow space. Conversely, control and AOC-treated defects demonstrated newly formed woven bone rich in cellular activity with no evidence of AOC remaining at the application site. Early healing appeared to be impaired by the presence of MFC and impeded by the presence of ORC. In contrast, AOC did not inhibit bone healing and suggest that AOC may be a better bone hemostatic material for procedures where bony fusion is critical and immediate hemostasis required.


Enhancing psychotherapy process with common factors feedback: A randomized, clinical trial.
In this study, we developed and tested a common factors feedback (CFF) system. The CFF system was designed to provide ongoing feedback to clients and therapists about client ratings of three common factors: (a) outcome expectations, (b) empathy, and (c) the therapeutic alliance. We evaluated the CFF system using randomized, clinical trial (RCT) methodology. Clients were 79 undergraduates who reported mild, moderate, or severe depressive symptoms at screening and pretreatment assessments. These clients were randomized to either: (a) treatment as usual (TAU) or (b) treatment as usual plus the CFF system (TAU + CFF). Both conditions entailed 5 weekly sessions of evidence-based therapy delivered by doctoral students in clinical psychology. Clients completed measures of common factors (i.e., outcome expectations, empathy, therapeutic alliance) and outcome at each session. Clients and therapists in TAU + CFF received feedback on client ratings of common factors at the beginning of Sessions 2 through 5. When surveyed, clients and therapists indicated that that they were satisfied with the CFF system and found it useful. Multilevel modeling revealed that TAU + CFF clients reported larger gains in perceived empathy and alliance over the course of treatment compared with TAU clients. No between-groups effects were found for outcome expectations or treatment outcome. These results imply that our CFF system was well received and has the potential to improve therapy process for clients with depressive symptoms. (PsycINFO Database Record


Narcolepsy onset is seasonal and increased following the 2009 H1N1 pandemic in China.
Narcolepsy is caused by the loss of hypocretin/orexin neurons in the hypothalamus, which is likely the result of an autoimmune process. Recently, concern has been raised over reports of narcolepsy in northern Europe following H1N1 vaccination. The study is a retrospective analysis of narcolepsy onset in subjects diagnosed in Beijing, China (1998-2010). Self-reported month and year of onset were collected from 629 patients (86% children). Graphical presentation, autocorrelations, chi-square, and Fourier analysis were used to assess monthly variation in onset. Finally, 182 patients having developed narcolepsy after October 2009 were asked for vaccination history. The occurrence of narcolepsy onset was seasonal, significantly influenced by month and calendar year. Onset was least frequent in November and most frequent in April, with a 6.7-fold increase from trough to peak. Studying year-to-year variation, we found a 3-fold increase in narcolepsy onset following the 2009 H1N1 winter influenza pandemic. The increase is unlikely to be explained by increased vaccination, as only 8 of 142 (5.6%) patients recalled receiving an H1N1 vaccination. Cross-correlation indicated a significant 5- to 7-month delay between the seasonal peak in influenza/cold or H1N1 infections and peak in narcolepsy onset occurrences. In China, narcolepsy onset is highly correlated with seasonal and annual patterns of upper airway infections, including H1N1 influenza. In 2010, the peak seasonal onset of narcolepsy was phase delayed by 6 months relative to winter H1N1 infections, and the correlation was independent of H1N1 vaccination in the majority of the sample.


Mutation Update of the CLCN5 Gene Responsible for Dent Disease 1.
Dent disease is a rare X-linked tubulopathy characterized by low molecular weight proteinuria, hypercalciuria, nephrocalcinosis and/or nephrolithiasis, progressive renal failure, and variable manifestations of other proximal tubule dysfunctions. It often progresses over a few decades to chronic renal insufficiency, and therefore molecular characterization is important to allow appropriate genetic counseling. Two genetic subtypes have been described to date: Dent disease 1 is caused by mutations of the CLCN5 gene, coding for the chloride/proton exchanger ClC-5; and Dent disease 2 by mutations of the OCRL gene, coding for the inositol polyphosphate 5-phosphatase OCRL-1. Herein, we review previously reported mutations (n = 192) and their associated phenotype in 377 male patients with Dent disease 1 and describe phenotype and novel (n = 42) and recurrent mutations (n = 24) in a large cohort of 117 Dent disease 1 patients belonging to 90 families. The novel missense and in-frame mutations described were mapped onto a three-dimensional homology model of the ClC-5 protein. This analysis suggests that these mutations affect the dimerization process, helix stability, or transport. The phenotype of our cohort patients supports and extends the phenotype that has been reported in smaller studies.


Salivary mucoepidermoid carcinoma: demonstration of transcriptionally active human papillomavirus 16/18.
Herein we test the following hypotheses: (1) High-risk Human Papillomavirus (HR-HPV) may be involved in the etiology of mucoepidermoid carcinoma (MEC), and (2) The detection rate of HR-HPV in MEC has been increasing over time. Ninety-eight archival MEC specimens from three institutions spanning three decades were studied for HPV16/18 E6/E7 transcripts. RNA was extracted from formalin-fixed paraffin embedded specimens and HPV16/18 E6/E7 expression assessed by nested reverse transcription polymerase chain reaction (RT-PCR). A subset of MEC were also studied for MECT1-MAML2 fusion transcripts by nested RT-PCR and amplicon sequencing. The HPV expression data was validated by immunofluorescence (IF) with monoclonal HPV16/18 E6 antibody, PCR with the GP5+/6+ consensus primers, and sequencing of RT-PCR amplicons. HPV genome was localized by in-situ hybridization with the Ventana Inform HPVIII Family 16 probe. P16(INK4a) overexpression and aberrant p53 expression were assessed by immunohistochemistry. HPV16 E6/E7 transcripts were demonstrated in (29/98) 30% of MEC by RT-PCR. HPV18 E6/E7 transcripts were demonstrated in 13/98 (13%) of MEC by RT-PCR. Seven of 98 tumors (7%) demonstrated both HPV16/18. No significant association was found between HPV status and gender, age, and tumor site. All 13 HPV18+ MEC were diagnosed between 2001 and 2010, whereas 45 MEC diagnosed from 1977 to 2000 were negative for HPV18 (p = 0.002). By contrast, there was no significant difference with respect to HPV16 detection and date of diagnosis. All MEC that were positive for E6 protein were also HPV16/18 positive by RT-PCR. Sequencing a subset of RT-PCR amplicons confirmed HPV type- and region-specific sequences. PCR using GP5+/6+ consensus primers demonstrated HPV status concordance in 9 of 10 cases. DNA degradation was present in the last case; the RT-PCR amplicons were sequenced from this case which confirmed the presence of HPV type- and region-specific sequences. Strong (+4/+4) and diffuse (>50%) nuclear and cytoplasmic p16 expression was seen in 64% of MEC in the glandular regions, and 18% of MEC in the solid, squamoid regions. No correlation was seen between p16 expression and HPV status. Twenty-nine MEC (22 HPV+ and 7 HPV-negative) were selected for further evaluation for p53 expression. Strong aberrant nuclear p53 expression was present in only 2/22 HPV + MEC (9%, both Grade 3); no HPV-negative MEC demonstrated aberrant p53 expression. MECT1-MAML2 fusion transcripts were demonstrated in 23/37 (62%) MEC. No significant association was found between the presence of the MECT1-MAML2 fusion transcripts and tumor grade, HPV status, gender, era of diagnosis (2000 and earlier vs. 2001-2010) or tumor site. We demonstrate for the first time that transcriptionally active HPV16/18 is common to MEC. These findings were validated by demonstrating concordant results by separate PCR with consensus primers, and/or confirming the presence of HPV type- and region-specific sequences in the RT-PCR amplicons. We also visualized E6 viral oncoprotein and HPV genome within tumor cells. HR-HPV is thus potentially implicated in the pathogenesis of MEC. The frequency of HPV18 detection is significantly increased in MEC diagnosed after 2001, whereas we found no differences in the HPV16 detection rates per era of diagnosis.


Basonuclin-null mutation impairs homeostasis and wound repair in mouse corneal epithelium.
At least two cellular processes are required for corneal epithelium homeostasis and wound repair: cell proliferation and cell-cell adhesion. These processes are delicately balanced to ensure the maintenance of normal epithelial function. During wound healing, these processes must be reprogrammed in coordination to achieve a rapid re-epithelialization. Basonuclin (Bnc1) is a cell-type-specific transcription factor expressed mainly in the proliferative keratinocytes of stratified epithelium (e.g., corneal epithelium, epidermis and esophageal epithelium) and the gametogenic cells in testis and ovary. Our previous work suggested that basonuclin could regulate transcription of ribosomal RNA genes (rDNA) and genes involved in chromatin structure, transcription regulation, cell-cell junction/communication, ion-channels and intracelllular transportation. However, basonuclin's role in keratinocytes has not been demonstrated in vivo. Here we show that basonuclin-null mutation disrupts corneal epithelium homeostasis and delays wound healing by impairing cell proliferation. In basonuclin-null cornea epithelium, RNA polymerase I (Pol I) transcription is perturbed. This perturbation is unique because it affects transcripts from a subset of rDNA. Basonuclin-null mutation also perturbs RNA polymerase II (Pol II) transcripts from genes encoding chromatin structure proteins histone 3 and HMG2, transcription factor Gli2, gap-junction protein connexin 43 and adheren E-cadherin. In most cases, a concerted change in mRNA and protein level is observed. However, for E-cadherin, despite a notable increase in its mRNA level, its protein level was reduced. In conclusion, our study establishes basonuclin as a regulator of corneal epithelium homeostasis and maintenance. Basonuclin likely coordinates functions of a subset of ribosomal RNA genes (rDNA) and a group of protein coding genes in cellular processes critical for the regulation of cell proliferation.


Sexual bother in men with advanced prostate cancer undergoing androgen deprivation therapy.
Men with advanced prostate cancer (APC) undergoing androgen deprivation therapy (ADT) often experience distressing sexual side effects. Sexual bother is an important component of adjustment. Factors associated with increased bother are not well understood. This study sought to describe sexual dysfunction and bother in APC patients undergoing ADT, identify socio-demographic and health/disease-related characteristics related to sexual bother, and evaluate associations between sexual bother and psychosocial well-being and quality of life (QOL). Baseline data of a larger psychosocial intervention study was used. Pearson's correlation and independent samples t-test tested bivariate relations. Multivariate regression analysis evaluated relations between sexual bother and psychosocial and QOL outcomes. The Expanded Prostate Cancer Index Composite sexual function and bother subscales, Center for Epidemiologic Studies Depression Scale, Functional Assessment of Cancer Therapy--General, and Dyadic Adjustment Scale were the main outcome measures. Participants (N = 80) were 70 years old (standard deviation [SD] = 9.6) and reported 18.7 months (SD = 17.3) of ADT. Sexual dysfunction (mean = 10.1; SD = 18.0) was highly prevalent. Greater sexual bother (lower scores) was related to younger age (β = 0.25, P = 0.03) and fewer months of ADT (β = 0.22, P = 0.05). Controlling for age, months of ADT, current and precancer sexual function, sexual bother correlated with more depressive symptoms (β = -0.24, P = 0.06) and lower QOL (β = 0.25, P = 0.05). Contrary to hypotheses, greater sexual bother was related to greater dyadic satisfaction (β = -0.35, P = 0.03) and cohesion (β = -0.42, P = 0.01). The majority of APC patients undergoing ADT will experience sexual dysfunction, but there is variability in their degree of sexual bother. Psychosocial aspects of sexual functioning should be considered when evaluating men's adjustment to ADT effects. Assessment of sexual bother may help identify men at risk for more general distress and lowered QOL. Psychosocial interventions targeting sexual bother may complement medical treatments for sexual dysfunction and be clinically relevant, particularly for younger men and those first starting ADT.


Managing HIV. Part 7: Professional issues. 7.4 HIV and complementary medicine.
Many people with HIV use complementary or alternative therapies, often without the knowledge of their doctors. To encourage open communication with their patients, doctors need to be aware of what complementary therapies are available and willing to discuss their use. They are then in a better position to defend their patients against exploitation or unsafe treatment.


The dynamics of T-cell receptor repertoire diversity following thymus transplantation for DiGeorge anomaly.
T cell populations are regulated both by signals specific to the T-cell receptor (TCR) and by signals and resources, such as cytokines and space, that act independently of TCR specificity. Although it has been demonstrated that disruption of either of these pathways has a profound effect on T-cell development, we do not yet have an understanding of the dynamical interactions of these pathways in their joint shaping of the T cell repertoire. Complete DiGeorge Anomaly is a developmental abnormality that results in the failure of the thymus to develop, absence of T cells, and profound immune deficiency. After receiving thymic tissue grafts, patients suffering from DiGeorge anomaly develop T cells derived from their own precursors but matured in the donor tissue. We followed three DiGeorge patients after thymus transplantation to utilize the remarkable opportunity these subjects provide to elucidate human T-cell developmental regulation. Our goal is the determination of the respective roles of TCR-specific vs. TCR-nonspecific regulatory signals in the growth of these emerging T-cell populations. During the course of the study, we measured peripheral blood T-cell concentrations, TCRbeta V gene-segment usage and CDR3-length spectratypes over two years or more for each of the subjects. We find, through statistical analysis based on a novel stochastic population-dynamic T-cell model, that the carrying capacity corresponding to TCR-specific resources is approximately 1000-fold larger than that of TCR-nonspecific resources, implying that the size of the peripheral T-cell pool at steady state is determined almost entirely by TCR-nonspecific mechanisms. Nevertheless, the diversity of the TCR repertoire depends crucially on TCR-specific regulation. The estimated strength of this TCR-specific regulation is sufficient to ensure rapid establishment of TCR repertoire diversity in the early phase of T cell population growth, and to maintain TCR repertoire diversity in the face of substantial clonal expansion-induced perturbation from the steady state.


Volumetric and two-dimensional image interpretation show different cognitive processes in learners.
In current practice, radiologists interpret digital images, including a substantial amount of volumetric images. We hypothesized that interpretation of a stack of a volumetric data set demands different skills than interpretation of two-dimensional (2D) cross-sectional images. This study aimed to investigate and compare knowledge and skills used for interpretation of volumetric versus 2D images. Twenty radiology clerks were asked to think out loud while reading four or five volumetric computed tomography (CT) images in stack mode and four or five 2D CT images. Cases were presented in a digital testing program allowing stack viewing of volumetric data sets and changing views and window settings. Thoughts verbalized by the participants were registered and coded by a framework of knowledge and skills concerning three components: perception, analysis, and synthesis. The components were subdivided into 16 discrete knowledge and skill elements. A within-subject analysis was performed to compare cognitive processes during volumetric image readings versus 2D cross-sectional image readings. Most utterances contained knowledge and skills concerning perception (46%). A smaller part involved synthesis (31%) and analysis (23%). More utterances regarded perception in volumetric image interpretation than in 2D image interpretation (Median 48% vs 35%; z = -3.9; P < .001). Synthesis was less prominent in volumetric than in 2D image interpretation (Median 28% vs 42%; z = -3.9; P < .001). No differences were found in analysis utterances. Cognitive processes in volumetric and 2D cross-sectional image interpretation differ substantially. Volumetric image interpretation draws predominantly on perceptual processes, whereas 2D image interpretation is mainly characterized by synthesis. The results encourage the use of volumetric images for teaching and testing perceptual skills.


Risk reduction practices in men who have sex with men in Switzerland: serosorting, strategic positioning, and withdrawal before ejaculation.
The aim of this study was to identify predictors of intentional use of the HIV risk reduction practices of serosorting, strategic positioning, and withdrawal before ejaculation during unprotected anal intercourse (UAI) with casual partners. A cross-sectional survey pertaining to the Swiss HIV behavioral surveillance system, using an anonymous self-administered questionnaire, was conducted in 2007 in a self-selected sample of men having sex with other men (MSM). Analysis was restricted to participants with UAI with casual partner(s) (N = 410). Logistic regression was used to estimate factors associated with intentional use of serosorting, strategic positioning, and withdrawal before ejaculation. In the previous 12 months, 71% of participants reported having UAI with a casual partner of different or unknown HIV-status. Of these, 47% reported practicing withdrawal, 38% serosorting, and 25% strategic positioning. In the 319 participants with known HIV-status, serosorting was associated with frequent Internet use to find partners (OR = 2.32), STI (OR = 2.07), and HIV testing in the past 12 months (OR = 1.81). Strategic positioning was associated with HIV-status (OR = 0.13) and having UAI with a partner of different or unknown HIV-status (OR = 3.57). Withdrawal was more frequently practiced by HIV-negative participants or participants reporting high numbers of sexual partners (OR = 2.48) and having UAI with a partner of unknown or different serostatus (OR = 2.08). Risk reduction practices are widely used by MSM, each practice having its own specificities. Further research is needed to determine the contextual factors surrounding harm reduction practices, particularly the strategic or opportunistic nature of their use.


Identification of very small open reading frames in the genomes of Holmes Jungle virus, Ord River virus, and Wongabel virus of the genus Hapavirus, family Rhabdoviridae.
Viruses of the family Rhabdoviridae infect a broad range of hosts from a variety of ecological and geographical niches, including vertebrates, arthropods, and plants. The arthropod-transmitted members of this family display considerable genetic diversity and remarkable genomic flexibility that enable coding for various accessory proteins in different locations of the genome. Here, we describe the genome of Holmes Jungle virus, isolated from Culex annulirostris mosquitoes collected in northern Australia, and make detailed comparisons with the closely related Ord River and Wongabel viruses, with a focus on identifying very small open reading frames (smORFs) in their genomes. This is the first systematic prediction of smORFs in rhabdoviruses, emphasising the intricacy of the rhabdovirus genome and the knowledge gaps. We speculate that these smORFs may be of importance to the life cycle of the virus in the arthropod vector.


Infectious toxicity of dexamethasone during all remission-induction chemotherapy: report of two cases and literature review.
Traditionally, children with acute lymphoblastic leukemia receive prednisone, as part of multiagent remission-induction chemotherapy. Recently, many cooperative groups use dexamethasone instead of prednisone during induction. We describe the infectious toxicities experienced by the first two patients in our institution treated with dexamethasone (10 mg/m(2)/day for 4 weeks with gradual tapering) during induction according to the dexamethasone arm of BFM 2000 and review the relevant literature that suggests an increased risk of infectious complications with dexamethasone. Only a prospective two-arm ALL dexamethasone study at two dose levels (6 and 10 mg/m(2)/day) will clarify if indeed the higher dose of dexamethasone during induction is more effective and without unacceptable toxicity.


An Integrative Review of Extended Use of Intrauterine Devices.
To explore the evidence on the impact of extended use of intrauterine devices (IUDs) use among women of reproductive age. Electronic resource databases used were PubMed, CINAHL, and Google Scholar. Peer-reviewed articles published during 2012 to 2022 were considered. Ten articles met the criteria and included data for a total of 7,420 women. Data were extracted from each study using the subheadings "Author/Date," "Participants," "Methods," "Interventions," and "Outcomes." Additionally, the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines were used to guide data extraction. Common themes were identified among each of the articles that supported the purpose of this integrative review, such as adverse effects, perceived effectiveness, pregnancy prevention, and cost effectiveness of extended use of IUDs. The available evidence supports the notion that extending IUD use beyond the original time frame approved by the U.S. Food and Drug Administration is safe and effective with minimal side effects. However, there remains a paucity of evidence examining this topic.