Fungal Isocyanide Synthases and Xanthocillin Biosynthesis in Aspergillus fumigatus.
Microbial secondary metabolites, including isocyanide moieties, have been extensively mined for their repertoire of bioactive properties. Although the first naturally occurring isocyanide (xanthocillin) was isolated from the fungus Penicillium notatum over half a century ago, the biosynthetic origins of fungal isocyanides remain unknown. Here we report the identification of a family of isocyanide synthases (ICSs) from the opportunistic human pathogen Aspergillus fumigatus Comparative metabolomics of overexpression or knockout mutants of ICS candidate genes led to the discovery of a fungal biosynthetic gene cluster (BGC) that produces xanthocillin (xan). Detailed analysis of xanthocillin biosynthesis in A. fumigatus revealed several previously undescribed compounds produced by the xan BGC, including two novel members of the melanocin family of compounds. We found both the xan BGC and a second ICS-containing cluster, named the copper-responsive metabolite (crm) BGC, to be transcriptionally responsive to external copper levels and further demonstrated that production of metabolites from the xan BGC is increased during copper starvation. The crm BGC includes a novel type of fungus-specific ICS-nonribosomal peptide synthase (NRPS) hybrid enzyme, CrmA. This family of ICS-NRPS hybrid enzymes is highly enriched in fungal pathogens of humans, insects, and plants. Phylogenetic assessment of all ICSs spanning the tree of life shows not only high prevalence throughout the fungal kingdom but also distribution in species not previously known to harbor BGCs, indicating an untapped resource of fungal secondary metabolism.IMPORTANCE Fungal ICSs are an untapped resource in fungal natural product research. Their isocyanide products have been implicated in plant and insect pathogenesis due to their ability to coordinate transition metals and disable host metalloenzymes. The discovery of a novel isocyanide-producing family of hybrid ICS-NRPS enzymes enriched in medically and agriculturally important fungal pathogens may reveal mechanisms underlying pathogenicity and afford opportunities to discover additional families of isocyanides. Furthermore, the identification of noncanonical ICS BGCs will enable refinement of BGC prediction algorithms to expand on the secondary metabolic potential of fungal and bacterial species. The identification of genes related to ICS BGCs in fungal species not previously known for secondary metabolite-producing capabilities (e.g., Saccharomyces spp.) contributes to our understanding of the evolution of BGC in fungi.


The encapsulation selectivity for anionic fission products imparted by an electride.
The nanoporous oxide 12CaO·7Al2O3 (C12A7) can capture large concentrations of extra-framework species inside its nanopores, while maintaining its thermodynamical stability. Here we use atomistic simulation to predict the efficacy of C12A7 to encapsulate volatile fission products, in its stoichiometric and much more effective electride forms. In the stoichiometric form, while Xe, Kr and Cs are not captured, Br, I and Te exhibit strong encapsulation energies while Rb is only weakly encapsulated from atoms. The high electronegativities of Br, I and Te stabilize their encapsulation as anions. The electride form of C12A7 shows a significant enhancement in the encapsulation of Br, I and Te with all three stable as anions from their atom and dimer reference states. Successive encapsulation of multiple Br, I and Te as single anions in adjacent cages is also energetically favourable. Conversely, Xe, Kr, Rb and Cs are unbound. Encapsulation of homonuclear dimers (Br2, I2 and Te2) and heteronuclear dimers (CsBr and CsI) in a single cage is also unfavourable. Thus, C12A7 offers the desirable prospect of species selectivity.


Utah's 72-Hour Waiting Period for Abortion: Experiences Among a Clinic-Based Sample of Women.
In 2012, Utah became the first state to enact a 72-hour waiting period for abortion. Despite debate about the law's potential effects, research has not examined women's experiences with it. A cohort of 500 women recruited at four family planning facilities in Utah in 2013-2014 completed baseline surveys at the time of an abortion information visit and follow-up telephone interviews three weeks later. Logistic regression and coding of open-ended responses were used to examine which women had abortions and, for those who did not, their reasons. Among the 309 women completing follow-up, 86% had had an abortion, 8% were no longer seeking abortion, 3% had miscarried or discovered they had not been pregnant, and 2% were still seeking abortion; one woman was still deciding, and the waiting period had pushed one woman beyond her facility's gestational limit for abortion. At the information visit, women reported little conflict about the abortion decision (mean score on a scale of 0-100 was 13.9 for those who eventually had an abortion and 28.5 for others). Low decisional conflict, but not socioeconomic status, was associated with having an abortion (odds ratio, 1.1). On average, eight days elapsed between the information visit and the abortion. As most women in this cohort were not conflicted about their decision when they sought care, the 72-hour waiting requirement seems to have been unnecessary. Individualized patient counseling for the small minority who were conflicted when they presented for care may have been more appropriate.


A comparison of treatment of canine cyclic hematopoiesis with recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF interleukin-3, and canine G-CSF.
Cyclic hematopoiesis in gray collie dogs is a stem cell disease in which abnormal regulation of cell production in the bone marrow causes cyclic fluctuations of blood cell counts. In vitro studies demonstrated that recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and granulocyte colony stimulating factor (G-CSF) all stimulated increases in colony formation by canine bone marrow progenitor cells. Based on these results, gray collie dogs were then treated with recombinant human (rh) GM-CSF, IL-3, or G-CSF subcutaneously to test the hypothesis that pharmacologic doses of one of these hematopoietic growth factors could alter cyclic production of cells. When recombinant canine G-CSF became available, it was tested over a range of doses. In vivo rhIL-3 had no effect on the recurrent neutropenia but was associated with eosinophilia, rhGM-CSF caused neutrophilia and eosinophilia but cycling of hematopoiesis persisted. However, rhG-CSF caused neutrophilia, prevented the recurrent neutropenia and, in the two animals not developing antibodies to rhG-CSF, obliterated periodic fluctuation of monocyte, eosinophil, reticulocyte, and platelet counts. Recombinant canine G-CSF increased the nadir neutrophil counts and amplitude of fluctuations at low doses (1 micrograms/kg/d) and eliminated all cycling of cell counts at high doses (5 and 10 micrograms/kg/d). These data suggest significant differences in the actions of these growth factors and imply a critical role for G-CSF in the homeostatic regulation of hematopoiesis.


Workplace bullying, working environment and health.
Improved work organisation could be of importance for decreased bullying in workplaces. Participants in the Swedish Longitudinal Occupational Survey of Health (SLOSH) responded to questions about work and workplace and whether they had been bullied during the past year in 2006. Those in worksites with at least five employees who did not report that they had been bullied in 2006 and without workplace change between 2006 and 2008 constituted the final sample (n=1,021 men and 1,182 women). Work characteristics and workplace factors in 2006 were used in multiple logistic regression as predictors of bullying in 2008. Separate analyses were performed for work characteristics and workplace factors respectively. Adjustments for demographic factors were made in all analyses. The question used for bullying was: "Are you exposed to personal persecution by means of vicious words or actions from your superiors or your workmates?" Such persecution any time during the past year was defined as bullying. For both genders organisational change and conflicting demands were identified as risk factors, and good decision authority as a protective factor. Dictatorial leadership, lack of procedural justice and attitude of expendability were male and lack of humanity a female risk factor for bullying.


Sirtuin 5 Regulates Proximal Tubule Fatty Acid Oxidation to Protect against AKI.
The primary site of damage during AKI, proximal tubular epithelial cells, are highly metabolically active, relying on fatty acids to meet their energy demands. These cells are rich in mitochondria and peroxisomes, the two organelles that mediate fatty acid oxidation. Emerging evidence shows that both fatty acid pathways are regulated by reversible posttranslational modifications, particularly by lysine acylation. Sirtuin 5 (Sirt5), which localizes to both mitochondria and peroxisomes, reverses post-translational lysine acylation on several enzymes involved in fatty acid oxidation. However, the role of the Sirt5 in regulating kidney energy metabolism has yet to be determined. We subjected male Sirt5-deficient mice (either +/- or -/-) and wild-type controls, as well as isolated proximal tubule cells, to two different AKI models (ischemia-induced or cisplatin-induced AKI). We assessed kidney function and injury with standard techniques and measured fatty acid oxidation by the catabolism of 14C-labeled palmitate to 14CO2. Sirt5 was highly expressed in proximal tubular epithelial cells. At baseline, Sirt5 knockout (Sirt5-/- ) mice had modestly decreased mitochondrial function but significantly increased fatty acid oxidation, which was localized to the peroxisome. Although no overt kidney phenotype was observed in Sirt5-/- mice, Sirt5-/- mice had significantly improved kidney function and less tissue damage compared with controls after either ischemia-induced or cisplatin-induced AKI. This coincided with higher peroxisomal fatty acid oxidation compared with mitochondria fatty acid oxidation in the Sirt5-/- proximal tubular epithelial cells. Our findings indicate that Sirt5 regulates the balance of mitochondrial versus peroxisomal fatty acid oxidation in proximal tubular epithelial cells to protect against injury in AKI. This novel mechanism might be leveraged for developing AKI therapies.


To kidney or not to kidney: Applying lessons learned from the simultaneous liver-kidney transplant policy to simultaneous heart-kidney transplantation.
As the medical community is increasingly offering transplantation to patients with increasing comorbidity burdens, the number of simultaneous heart-kidney (SHK) transplants is rising in the United States. How to determine eligibility for SHK transplant versus heart transplant alone is unknown. In this review, we situate this problem in the broader picture of organ shortage. We critically appraise available literature on outcomes in SHK versus heart transplant alone. We posit staged kidney-after-heart transplantation as a plausible alternative to SHK transplantation and review the pros and cons. Drawing lessons from the field of simultaneous liver-kidney transplant, we argue for an analogous policy for SHK transplant with standardized minimal eligibility criteria and a modified Safety Net provision. The new policy will serve as a starting point for comparing simultaneous versus staged approaches and refining the medical eligibility criteria for SHK.


A Case of Guillain-Barré Syndrome Associated With COVID-19.
A novel member of human RNA coronavirus, which is an enveloped betacoronavirus, has been termed severe acute respiratory syndrome coronavirus-2 (SARS COV-2). The illness caused by SARS COV-2 is referred to as the coronavirus disease 2019 (COVID-19). It is a highly contagious disease that has resulted in a global pandemic. The clinical spectrum of COVID-19 ranges from asymptomatic illness to acute respiratory distress syndrome, septic shock, multi-organ dysfunction, and death. The most common symptoms include fever, fatigue, dry cough, dyspnea, and diarrhea. Neurological manifestations have also been reported. However, the data on the association of Guillain-Barré syndrome (GBS) with COVID-19 are scarce. We report a rare case of a COVID-19-positive 36-year-old immunocompromised male who presented with clinical features of GBS. His clinical examination showed generalized weakness and hyporeflexia. The cerebrospinal fluid (CSF) analysis showed albuminocytological dissociation. Intravenous immunoglobulin (IVIG) was administered based on the high clinical suspicion of GBS. The patient's neurological condition worsened with progression to bulbar weakness and ultimately neuromuscular respiratory failure requiring mechanical ventilation. His nerve conduction studies were consistent with demyelinating polyneuropathy. He received five plasma exchange treatments and was successfully weaned from mechanical ventilation. A brain and cervical spine magnetic resonance imaging was obtained to rule out other causes, which was normal. COVID-19 is believed to cause a dysregulated immune system, which likely plays an important role in the neuropathogenesis of GBS.


Cost-effectiveness of catheter ablation for rhythm control of atrial fibrillation.
Objective. The objective of this study is to evaluate the cost-effectiveness of catheter ablation for rhythm control compared to antiarrhythmic drug (AAD) therapy in patients with atrial fibrillation (AF) who have previously failed on an AAD. Methods. An economic model was developed to compare (1) catheter ablation and (2) AAD (amiodarone 200 mg/day). At the end of the initial 12 month phase of the model, patients are classified as being in normal sinus rhythm or with AF, based on data from a meta-analysis. In the 5-year Markov phase of the model, patients are at risk of ischemic stroke each 3-month model cycle. Results. The model estimated that, compared to the AAD strategy, ablation had $8,539 higher costs, 0.033 fewer strokes, and 0.144 more QALYS over the 5-year time horizon. The incremental cost per QALY of ablation compared to AAD was estimated to be $59,194. The probability of ablation being cost-effective for willingness to pay thresholds of $50,000 and $100,000 was estimated to be 0.89 and 0.90, respectively. Conclusion. Based on current evidence, pulmonary vein ablation for treatment of AF is cost-effective if decision makers willingness to pay for a QALY is $59,194 or higher.


Rat renal cortical slices demonstrate p-aminohippurate/glutarate exchange and sodium/glutarate coupled p-aminohippurate transport.
In isolated basolateral membrane vesicles p-aminohippurate (PAH) transport may be coupled indirectly to the sodium gradient through PAH/glutarate [or alpha-ketoglutarate (alpha-KG)] exchange and Na/glutarate cotransport. In this study, rat renal cortical slices were used to examine indirect coupling of PAH transport to sodium in intact renal tissue. Like basolateral membrane vesicles, slices demonstrated avid uptake of 50 microM [14C]glutarate. Steady-state tissue/medium ratios of 30 were achieved by 90 to 120 min. Uptake was inhibited markedly by lithium and fumarate. PAH also inhibited glutarate accumulation, but through acceleration of glutarate efflux i.e., PAH/glutarate exchange, rather than direct inhibition of uptake. PAH-driven efflux of glutarate from slices was blocked by probenecid, which inhibits PAH/glutarate exchange in vesicles. Inasmuch as slices showed both Na/glutarate uptake and PAH/glutarate exchange, externally added glutarate should stimulate PAH uptake in slices. Indeed, in the presence of sodium, 50 microM external glutarate approximately doubled PAH accumulation by the slices. Stimulation by glutarate was abolished by either lithium or fumarate, or by elimination of sodium from the external buffer. The stimulatory effect was specific for glutarate or alpha-KG. Acetate, fumarate and succinate stimulated PAH uptake poorly, if at all. Neither fumarate nor lithium was able to eliminate concentrative PAH uptake completely, suggesting that a portion of PAH transport may occur without Na-dependent glutarate or alpha-KG recycling, i.e., that it may run directly off metabolic alpha-KG production or via a completely glutarate/alpha-KG independent mechanism.


[The role of the protein with molecular weight of 22 kD in the phagocytosis of vaccine strain of Yersinia pestis EV].
The comparative study of different stages of the phagocytosis of vaccine strain Y. pestis EV and its achromogenic variants (AV), has been carried out with the use of peritoneal macrophages as an in vitro experimental model. As revealed in this study, AV whose outer membrane contains no protein with a molecular weight of 22 kD exhibit lower capacity for adherence and for being ingested by phagocytes than the initial strain. The absence of this protein does not inhibit the multiplication of AV inside phagocytes, leading to incomplete phagocytosis, which is characteristic of the initial strain. The suggestion is made that the 22 kD protein may be one of the adhesion factors, necessary for ensuring the initial stages of phagocytosis.


Head Growth Trajectory and Neurodevelopmental Outcomes in Preterm Neonates.
To evaluate the association between head growth (HG) during neonatal and postdischarge periods and neurodevelopmental outcomes of preterm neonates of <29 weeks gestational age. We conducted a retrospective cohort study of infants <29 weeks gestational age admitted between 2009 and 2011 to participating Canadian Neonatal Network units and followed by Canadian Neonatal Follow-Up Network clinics. Differences in head circumference (ΔHC) z score were calculated for 3 time periods, which include admission to discharge, discharge to follow-up at 16-36 months, and admission to follow-up. These were categorized in 1 reference group (ΔHC z score between -1 and +1) and 4 study groups (ΔHC z score of <-2, between -2 to -1, +1 to +2, and >+2). Neurodevelopmental outcomes were compared with the reference group. 1973 infants met the inclusion criteria. Poor HG occurred frequently during the NICU admission (ΔHC z score <-2 in 24% infants versus 2% infants post-discharge) with a period of "catch-up" growth postdischarge. Significant neurodevelopmental impairment was higher in infants with the poorest HG from admission to follow-up (adjusted odds ratio 2.18, 95% confidence interval 1.50-3.15), specifically cognitive and motor delays. Infants with poor initial HG and catch-up postdischarge have a lower adjusted odds ratio of significant neurodevelopmental impairment (0.35, 95% CI 0.16-0.74). Infants with poor HG received a longer duration of parenteral nutrition and mechanical ventilation and had poor weight gain. Poor HG during the neonatal and postdischarge periods was associated with motor and cognitive delays at 16 to 36 months.


Exploiting redundancy in large materials datasets for efficient machine learning with less data.
Extensive efforts to gather materials data have largely overlooked potential data redundancy. In this study, we present evidence of a significant degree of redundancy across multiple large datasets for various material properties, by revealing that up to 95% of data can be safely removed from machine learning training with little impact on in-distribution prediction performance. The redundant data is related to over-represented material types and does not mitigate the severe performance degradation on out-of-distribution samples. In addition, we show that uncertainty-based active learning algorithms can construct much smaller but equally informative datasets. We discuss the effectiveness of informative data in improving prediction performance and robustness and provide insights into efficient data acquisition and machine learning training. This work challenges the "bigger is better" mentality and calls for attention to the information richness of materials data rather than a narrow emphasis on data volume.


Central nervous system manifestations of tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a neurocutaneous autosomal-dominant genetic syndrome marked by development of hamartomatous lesions arising from dysfunction of the mammalian target of rapamycin (mTOR) pathway. Although TSC remains a heterogeneous clinical entity, the recent inclusion of genetic diagnostic criteria reflects advancement in our understanding of its underlying etiopathogenesis. Abnormal cellular growth, differentiation, and migration result in multisystem sequelae, with neurologic manifestations of TSC representing the primary cause of morbidity and mortality for the majority of individuals. Modern imaging techniques aid in the diagnosis of TSC and guide treatment strategies by revealing central nervous system findings. Cortical tubers are the namesake lesion of the disorder and occur in up to 90% of cases, often exerting significant epileptogenic potential. Subependymal nodules are found in 80% of patients as calcified tumors lining the ependyma of the lateral ventricles. In some cases, these nodules are thought to progress to subependymal giant cell astrocytomas and may present with obstructive hydrocephalus. Retinal astrocytic hamartomas are also common, present in 50% of patients. Surgery remains the treatment of choice for large or symptomatic lesions, though clinical trials have highlighted a potential role for mTOR pathway antagonism. A multidisciplinary approach is necessary for achieving optimal patient outcomes.


Visual Outcomes, Endothelial Loss, and Complications Following Iris-Claw Intraocular Lens Implantation and Trocar-Assisted Sutureless Scleral Fixation of Intraocular Lenses.
Iris-claw intraocular lens (IC-IOL) implantation and sutureless scleral fixation of intraocular lenses (SSF-IOL) are two commonly preferred surgical approaches for the management of aphakic patients without sufficient capsular support. The aim of this study was to compare the outcomes of IC-IOL implantation and trocar-assisted SSF-IOL implantation. The medical records of secondary IOL implantation patients were retrospectively reviewed. All patients had a detailed ophthalmological examination, including LogMAR best-corrected distance visual acuity (CDVA), intraocular pressure (IOP), and endothelial cell density (ECD) preoperatively and postoperatively. SPSS 21.0 software was used for the statistical analysis. There were 15 patients in the IC-IOL group and 12 patients in the SSF-IOL group. Age and gender distributions were similar between the groups (p=0.456 and p=0.398, respectively). Similarly, patients in both groups had similar CDVA preoperatively and postoperatively (p=0.51, p=0.48, respectively). Both IC-IOL and SSF-IOL implantation significantly increased CDVA (p=0.001 and p=0.005, respectively). IOP remained unchanged in both groups. However, ECD reduced significantly following both IC-IOL and SSF-IOL implantation (p=0.001 and p=0.005, respectively) and trocar-assisted SSF-IOL implantation resulted in significantly more endothelial loss compared to IC-IOL implantation (439.5±89 vs. 164.4±53, p=0.013). Both surgical approaches increased CDVA significantly and at similar levels. However, trocar-assisted SSF-IOL implantation resulted in significantly more endothelial loss compared to IC-IOL implantation. None of the patients developed bullous keratopathy, but this difference should be kept in mind, especially in patients with critically low ECD.


Risk factors for father-daughter incest: data from an anonymous computerized survey.
Retrospective data from 2,034 female participants, provided anonymously using a computer-assisted self-interview, were used to identify risk factors for father-daughter incest (FDI). A total of 51 participants had reported having experienced FDI. The risk factors identified within the nuclear family by the multiple logistic regression analysis included the following: (a) Having parents whose relationship included verbal or physical fighting or brutality increased the likelihood of FDI by approximately 5 times; (b) families accepting father-daughter nudity as measured by a scale with values ranging from 0 to 4 increased the likelihood of FDI by approximately 2 times for each unit value increase of 1 above 0; (c) demonstrating maternal affection protected against FDI. The likelihood of being a victim of FDI was highest if the participant's mother never kissed or hugged her; it decreased by 0.44 for a 1-unit increase in affection and by 0.19 times for a 2-unit increase; and (d) being in homes headed by single-parent mothers or where divorce or death of the father had resulted in a man other than the biological father living in the home increased the risk of FDI by approximately 3.2 times. The results were consistent with the idea that FDI in many families was the cumulative result of a circular pattern of interactions, a finding that has implications for treatment of the perpetrator, the victim, and the families. The data also suggested it may be possible to design an information program for parents that will result in reducing the risk of FDI in families implementing the program's recommendations.


Optimized canine dental computed tomographic protocol in medium-sized mesaticepahlic dogs.
The purpose of this study was to develop an optimized imaging protocol for canine dental imaging using single-detector row CT and multidetector row CT. Two medium-sized mesaticephalic cadaver dog heads were scanned using varying slice thickness, sequential vs. helical scanning mode, sequential CT table increment, pitch, image reconstruction algorithm, and an additional moderate edge enhancement filter. For each series, two reviewers scored dental roots as not visible (score: 0), partially visible (score: 1) or completely visible (score: 2). Sharpness of the dento-alveolar margin was scored as blurry (score: 0), sharp (score: 1) or very sharp (score: 2). Consensus reader scores were compared among CT protocols. For single-detector row CT, complete visualization of all tooth roots was only achieved with 1 mm slice thickness and image interval sequential or helical series (pitch of 1 or 2) using a high-frequency image reconstruction algorithm with or without additional edge enhancement. For four-detector row CT, complete visibility of all tooth roots was recorded in all series. For single-detector row CT, all medium frequency algorithm series yielded blurry margin scores and all other series yielded sharp margin scores. For four-detector row CT, sequential and helical 0.5 mm thickness images yielded very sharp dento-alveolar margin scores, whereas the 1 mm series yielded only sharp margin scores. Authors concluded that the optimal dental imaging protocol was a sequential mode 1 mm slice thickness and interval with a high-frequency image reconstruction algorithm and an additional moderate edge enhancement filter.


DcardNet: Diabetic Retinopathy Classification at Multiple Levels Based on Structural and Angiographic Optical Coherence Tomography.
Optical coherence tomography (OCT) and its angiography (OCTA) have several advantages for the early detection and diagnosis of diabetic retinopathy (DR). However, automated, complete DR classification frameworks based on both OCT and OCTA data have not been proposed. In this study, a convolutional neural network (CNN) based method is proposed to fulfill a DR classification framework using en face OCT and OCTA. A densely and continuously connected neural network with adaptive rate dropout (DcardNet) is designed for the DR classification. In addition, adaptive label smoothing was proposed and used to suppress overfitting. Three separate classification levels are generated for each case based on the International Clinical Diabetic Retinopathy scale. At the highest level the network classifies scans as referable or non-referable for DR. The second level classifies the eye as non-DR, non-proliferative DR (NPDR), or proliferative DR (PDR). The last level classifies the case as no DR, mild and moderate NPDR, severe NPDR, and PDR. We used 10-fold cross-validation with 10% of the data to assess the network's performance. The overall classification accuracies of the three levels were 95.7%, 85.0%, and 71.0% respectively. A reliable, sensitive and specific automated classification framework for referral to an ophthalmologist can be a key technology for reducing vision loss related to DR.


Bayesian risk assessment model of human cryptosporidiosis cases following consumption of raw Eastern oysters (Crassostrea virginica) contaminated with Cryptosporidium oocysts in the Hillsborough River system in Prince Edward Island, Canada.
Cryptosporidium spp. has been associated with foodborne infectious disease outbreaks; however, it is unclear to what extent raw oyster consumption poses a risk to public health. Control of Cryptosporidium in shellfish harvest seawater in Canada is not mandatory and, despite relay/depuration processes, the parasite can remain viable in oysters for at least a month (depending on initial loads and seawater characteristics). Risks of human infection and illness from exposure to oysters contaminated with Cryptosporidium oocysts were assessed in a Bayesian framework. Two data sets were used: counts of oocysts in oysters harvested in Approved, Restricted, and Prohibited zones of the Hillsborough River system; and oocyst elimination rate from oysters exposed to oocysts in laboratory experiments. A total of 20 scenarios were assessed according to number of oysters consumed in a single serving (1, 10 and 30) and different relay times. The median probability of infection and developing cryptosporidiosis (e.g. illness) due to the consumption of raw oysters in Prince Edward Island was zero for all scenarios. However, the 95th percentiles ranged from 2% to 81% and from 1% to 59% for probability of infection and illness, respectively. When relay times were extended from 14 to 30 days and 10 oysters were consumed in one serving from the Restricted zones, these probabilities were reduced from 35% to 16% and from 15% to 7%, respectively. The 14-day relay period established by Canadian authorities for harvesting in Restricted zones seems prudent, though insufficient, as this relay period has been shown to be enough to eliminate fecal coliforms but not Cryptosporidium oocysts, which can remain viable in the oyster for over a month. Extending relay periods of 14 and 21 days for oysters harvested in Restricted zones to 30 days is likely insufficient to substantially decrease the probability of infection and illness. The highest risk was found for oysters that originated in Prohibited zones. Our findings suggest that Cryptosporidium oocysts are a potential cause of foodborne infection and illness when consuming raw oysters from Hillsborough River, one of the most important oyster production bays on Prince Edward Island. We discuss data gaps and limitations of this work in order to identify future research that can be used to reduce the uncertainties in predicted risks.


Activation of resting T lymphocytes by anti-CD3 (T3) antibodies in the absence of monocytes.
The antigen receptor molecules on human T lymphocytes are noncovalently associated on the cell surface with the CD3 (T3) molecular complex. Perturbation of this complex with anti-CD3 monoclonal antibodies induces T cell activation. Previous studies have demonstrated that this process requires the participation of monocytes. In the present report, we demonstrate that purified, resting (G0 phase) T cells incubated with monoclonal anti-CD3 antibodies proliferate in response to purified interleukin 2 (IL 2), in a lymphokine dose-dependent fashion. Anti-CD3 antibody or IL 2 alone did not trigger cell division. The effect was specific for anti-CD3 antibodies because monoclonal antibodies reactive with other surface molecules (OKT4, OKT8, L368) were inactive. Furthermore, the same phenomenon was observed when anti-CD3 antibody Leu-4 (IgG1) was incubated with cells of individuals whose monocytes cannot process antibodies of the IgG1 subclass (Leu-4 nonresponders). In addition, both F(ab')2 and Fab fragments of anti-CD3 antibody OKT3 were also capable of rendering T cells receptive to the IL 2 growth signal. These data indicate that neither monocytes nor CD3 receptor cross-linking are required absolutely for resting T cell activation, provided that IL 2 is supplied exogenously. T lymphocytes treated with anti-CD3 antibodies proliferated in response to both purified mitogen-induced and recombinant IL 2. Antibodies to the IL 2 receptor (anti-Tac) inhibited the proliferation. Thus, the most likely mechanism for anti-CD3 antibody-mediated triggering is induction of IL 2 receptors.


One-week dual therapy with ranitidine bismuth citrate and clarithromycin for the treatment of Helicobacter pylori infection in Brazilian patients with peptic ulcer.
To assess the efficacy and safety of ranitidine bismuth citrate plus clarithromycin given for 1 wk in Brazilian patients with peptic ulcer. One hundred and twenty patients with peptic ulcer were randomized in two treatment groups: (1) 1-wk regimen consisting of ranitidine bismuth citrate 400 mg b.i.d. with clarithromycin 500 mg b.i.d. or (2) 2-wk regimen of the same treatment. Eradication of the infection was considered when both the histologic examination and the urease test were negative for the infection 3 mo after treatment. By intention to treat analysis, Helicobacter pylori (H pylori) was eradicated in 73% and 76% of patients, respectively treated for 1 or 2 wk (P>0.05). By per protocol analysis, the eradication rates were 80% and 83%, respectively, in patients treated for 1 or 2 wk (P>0.05). Nine patients (8.2%) reported minor side effects. One-week therapy with ranitidine bismuth citrate and clarithromycin is safe, well tolerated and effective for treatment of H pylori infection, and appears to be comparable to the 2-wk regimen in terms of efficacy.


Trauma, depression, and resilience of earthquake/tsunami/nuclear disaster survivors of Hirono, Fukushima, Japan.
A mega-earthquake and tsunami struck the northeastern coast of Japan, and many survivors were forced to evacuate to temporary housing due to rising radiation levels. The aims of this study were to investigate the prevalence of symptoms of post-traumatic stress disorder (PTSD), depression, and poor general health among survivors, to test the predictive roles of resilience on mental and physical health, and to examine the predictive sociodemographic factors on resilience. Two hundred and forty-one evacuees (men/women: 116/125) from Hirono, Fukushima participated in the study. They were asked to complete the Connor-Davidson Resilience Scale, the Zung Self-Rating Depression Scale, the Impact of Events Scale-Revised, and a demographic questionnaire. Among all participants, 53.5% exhibited the clinically concerning symptoms of PTSD, and among them 33.2% indicated clinical PTSD symptoms. Additionally, 66.8% reported symptoms of depression, and among them 33.2% showed mildly depressive symptoms, while 19.1% and 14.5% demonstrated moderate and severe depressive symptoms, respectively. Resilience was a significant buffer for depression, PTSD, and general health. Additionally, employment status, eating/exercise habits, and drinking habits predicted resilience. The results indicated that depression and PTSD are prevalent among the survivors of massive earthquakes, tsunamis, and accidents from nuclear power plants. However, the results also showed that some survivors managed to endure the traumatic events relatively well, and resilience was a significant protective factor in dealing with such events. Therefore, it is crucial to assist survivors in improving their resilience by providing job opportunities and encouraging a healthy lifestyle.


Methamphetamine and paranoia: the methamphetamine experience questionnaire.
Paranoia in methamphetamine (MA) users is not well characterized or understood. To investigate this phenomenon, we created the Methamphetamine Experience Questionnaire (MEQ), and tested its reliability and validity in assessing MA-induced paranoia. We administered the MEQ to 274 MA-dependent subjects. Of the total subjects, 45% (123) first experienced paranoia with MA use; 55% did not. Obtaining or using a weapon while paranoid was common (37% and 11% of subjects with MA-induced paranoia, respectively). Test-retest and inter-rater reliability for MA-induced paranoia showed substantial agreement (kappa = .77, p < .05 and kappa = .80, p < .05, respectively). First episodes of paranoia occurred more often with intravenous use of MA, and subsequent episodes at higher doses. There was modest correlation between paranoia on the MEQ and the Brief Symptom Inventory (BSI) paranoid ideation scale (rho = .27, p < .05). As expected, there was a poor correlation between paranoia on the MEQ and the BSI depression scale (rho = .14, p = .07). The MEQ provides useful information on drug use variables that contribute to paranoia commonly associated with MA use. (Am J Addict 2010;00:1-14).


Short-term aromatase-enzyme blockade unmasks impaired feedback adaptations in luteinizing hormone and testosterone secretion in older men.
The mechanisms subserving hypoandrogenemia and relative hypogonadotropism in older men are not known. The present study tests the clinical hypothesis that aging impairs hypothalamopituitary adaptations to feedback withdrawal induced by antagonism of estrogen biosynthesis. To this end, we appraised gonadal axis responses to estrogen depletion induced by anastrozole (a potent and selective aromatase inhibitor) in nine older and 11 young men vs. placebo in 17 other older and eight young men. The study design comprised a prospectively randomized, double-blind, parallel-cohort intervention. To monitor LH release, blood was sampled every 10 min for 24 h; LH concentrations were assayed by two-site monoclonal immunoradiometric assay; pulsatile LH release quantitated by a model-free discrete peak-detection technique (Cluster); feedback-dependent orderliness of LH secretion via the approximate entropy statistic; and 24-h rhythmicity of LH concentrations by cosine analysis. At baseline, older men had comparable estradiol and testosterone but lower LH concentrations than young controls. Exposure to anastrozole reduced (24-h pooled) serum estradiol concentrations by 50% (P < 0.001) and elevated mean LH concentrations by 2.1-fold (P < 0.001) in both the young and older cohorts. However, older men failed to achieve young adult augmentation of the following: 1) total testosterone concentrations (P < 0.01) or molar testosterone to SHBG ratios (P < 0.01); 2) incremental LH pulse amplitude (P < 0.001) and LH peak area (P < 0.01); 3) mean LH pulse frequency (P = 0.0044); and 4) quantifiable irregularity (approximate entropy) of LH release patterns (P < 0.001). FSH concentrations became comparable in the two age cohorts. In summary, administration of a potent and selective aromatase antagonist reduces estradiol and elevates mean LH concentrations equivalently in young and older men. The low estrogen-feedback state in elderly men unmasks diminished incremental LH pulse amplitude and area; absence of further acceleration of LH pulse frequency; impaired regulation of the orderliness of LH release; and reduced testosterone to SHBG ratios. Thus, aging alters expected hypothalamopituitary-gonadal adaptations to short-term partial estrogen depletion in healthy men.


UBE2M Is a Stress-Inducible Dual E2 for Neddylation and Ubiquitylation that Promotes Targeted Degradation of UBE2F.
UBE2M and UBE2F are two family members of neddylation E2 conjugating enzyme that, together with E3s, activate CRLs (Cullin-RING Ligases) by catalyzing cullin neddylation. However, whether and how two E2s cross-talk with each other are largely unknown. Here, we report that UBE2M is a stress-inducible gene subjected to cis-transactivation by HIF-1 and AP1, and MLN4924, a small molecule inhibitor of E1 NEDD8-activating enzyme (NAE), upregulates UBE2M via blocking degradation of HIF-1α and c-JUN. UBE2M is a dual E2 for targeted ubiquitylation and degradation of UBE2F, acting as a neddylation E2 to activate CUL3-Keap1 E3 under physiological conditions but as a ubiquitylation E2 for Parkin-DJ-1 E3 under stressed conditions. UBE2M-induced UBE2F degradation leads to CRL5 inactivation and subsequent NOXA accumulation to suppress the growth of lung cancer cells. Collectively, our study establishes a negative regulatory axis between two neddylation E2s with UBE2M ubiquitylating UBE2F, and two CRLs with CRL3 inactivating CRL5.


Ewing's sarcoma family tumors are sensitive to tumor necrosis factor-related apoptosis-inducing ligand and express death receptor 4 and death receptor 5.
In this study, we investigated the sensitivity of Ewing's sarcoma family tumors (ESFTs) of children and adolescents to the tumor necrosis factor-related apoptosis-inducing Ligand (TRAIL). TRAIL binds to death receptors (DRs) DR4, DR5, DcR1, and DcR2. Either DR4 or DR5 can induce apoptosis, whereas DcR1 and DcR2 are considered inhibitory receptors. Nine of 10 ESFT cell lines, including several that were Fas resistant, underwent apoptosis with TRAIL through activation of caspase-10, capase-8 (FLICE), caspase-3, and caspase-9. In contrast to the Fas signaling pathway, caspase-10, but not caspase-8 or the Fas-associated death domain-containing molecule, was recruited to the TRAIL receptor-associated signaling complex. We found that 9 of 10 ESFT cell lines expressed both DR4 and DR5 by Western blotting, whereas the TRAIL-resistant line expressed only DR4. However, DR4 was absent from the cell surface in the resistant and two additional lines (three of five tested lines), suggesting that it may have been nonfunctional. On the contrary, DR5 was located on the cell surface in all four sensitive lines tested, being absent only from the cell surface of the resistant line that was also DR5-negative by Western blotting. In agreement with these findings, the resistance of the line was overcome by restoration of DR5 levels by transfection. Levels of DcR1 and DcR2 or levels of the FLICE-inhibitory protein (FLIP) did not correlate with TRAIL resistance, and protein synthesis inhibition did not sensitize the TRAIL-resistant line to TRAIL. Because these data suggested that sensitivity of ESFTs to TRAIL was mainly based on the presence of DR4/DR5, we investigated the presence of these receptors in 32 ESFT tissue sections by immunohistochemistry. We found that 23 of 32 tumor tissues (72%) expressed both receptors, 8 of 32 (25%) expressed one receptor only, and 1 was negative for both. Our finding of wide expression of DR4/DR5 in ESFT in vivo, in combination with their high sensitivity to TRAIL in vitro and the reported lack of toxicity of TRAIL in mice and monkeys, suggests that TRAIL may be a novel effective agent in the treatment of ESFTs.


Association between physical function and onset of coronary heart disease in a cohort of community-dwelling older populations: The SONIC study.
Physical function is a strong predictor of the adverse outcomes of cardiovascular disease in older populations. However, studies of healthy older people on the prevention of coronary heart disease (CHD) are very limited. We prospectively examined the association of walking speed and handgrip strength with CHD in the community-dwelling older populations. The study cohort in Japan included 1272 older people free from heart disease at the baseline. Physical function was identified based on walking speed and handgrip strength assessment at the survey site. Any new case of CHD was identified based on a self-reported doctor's diagnosis. Cox-proportion hazard models were adjusted for covariate factors to examine the CHD risk. During the 7-year follow-up, 45 new cases of CHD (25 men and 20 women) were documented. Slow walking speed was strongly associated with CHD risk after adjusting for all confounding factors in the total participants and women (hazard ratio (HR)= 2.53, 95%confidence interval (CI), 1.20-5.33, p=0.015, and HR= 4.78, 95% CI,1.07-21.35, p=0.040, respectively), but not in men. Weak grip strength was associated with CHD after age-adjustment (HR= 2.45, 95%CI, 1.03-5.81, p=0.043) only in men. However, after additional multivariate adjustment, the associations were getting weaker.


Whole genome sequencing reveals candidate genes involving in PAS resistance in M. Tuberculosis isolated from patients in Thailand.
Para-amino salicylic acid (PAS) was first reported by Lehmann in 1946 and used for tuberculosis treatment. However, due to its adverse effects, it is now used only as a second line anti-tuberculosis drug for treatment of multidrug resistant or extensively drug resistant M. tuberculosis. The structure of PAS is similar to para-amino benzoic acid (pABA), an intermediate metabolite in the folate synthesis pathway. The study has identified mutations in genes in folate pathway and their intergenic regions for their possibilities in responsible for PAS resistance. Genomic DNA from 120 PAS-resistant and 49 PAS-sensitive M. tuberculosis isolated from tuberculosis patients in Thailand were studied by whole genome sequencing. Twelve genes in the folate synthesis pathway were investigated for variants associated with PAS resistance. Fifty-one SNVs were found in nine genes and their intergenic regions (pabC, pabB, folC, ribD, thyX, dfrA, thyA, folK, folP). Functional correlation test confirmed mutations in RibD, ThyX, and ThyA are responsible for PAS resistance. Detection of mutation in thyA, folC, intergenic regions of thyX, ribD, and double deletion of thyA dfrA are proposed for determination of PAS resistant M. tuberculosis.


Differential distribution of the cognate and heat-stress-induced isoforms of high Mr cis-trans prolyl peptidyl isomerase (FKBP) in the cytoplasm and nucleoplasm.
Wheat root tips express a 73 kDa cognate isoform and a 77 kDa heat-shock-induced isoform of peptidyl prolyl cis-trans isomerase (FK506 binding protein; FKBP) that is part of a chaperone complex with hsp90. The 73 kDa and 77 kDa FKBPs have very similar sequences, differing primarily in the N- and C-terminal 20 amino acids. In order to define the potential functional roles of these proteins, the 73 kDa and 77 kDa FKBPs were localized in root tips using antigen-affinity purified antibodies as a probe. The cognate 73 kDa FKBP is localized in the cytoplasm and appears enriched around the periphery of the early vacuole and vesicles exiting the trans-Golgi. Parallel assays with antibodies directed against tonoplast aquaporin and pyrophosphatase confirmed the association of FKBP with an early vacuole compartment. Sucrose gradient centrifugation analysis of root tip lysates also showed that 73 kDa FKBP is co-fractionated with tonoplast aquaporin and V-ATPase in a light compartment near the top of the gradient. Heat-shock treatment of root tips induces the accumulation of 77 kDa FKBP while the abundance of 73 kDa FKBP remains constant. Quantitative EM immunogold assays of the intracellular distribution of FKBP over an 8 h heat-shock time-course showed that FKBP is initially present in the cytoplasm, but is transported into the nucleus where it accumulates in the nucleoplasm and into specific subnuclear domains. The results of this study show that the intracellular distribution of the high Mr FKBPs in wheat root tips differs at normal and elevated temperatures, indicating different functional roles for the FKBP isoforms.


Determination of chitin content in fungal cell wall: an alternative flow cytometric method.
The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted GlycosylPhosphatidylInositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P < 0.001) than chs3Δ/chs3Δ and pga31Δ/Δ especially in the presence of caspofungin. Ca. parapsilosis, Ca. tropicalis, and Ca. albicans showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi.


Exocytosis and endocytosis: modes, functions, and coupling mechanisms.
Vesicle exocytosis releases content to mediate many biological events, including synaptic transmission essential for brain functions. Following exocytosis, endocytosis is initiated to retrieve exocytosed vesicles within seconds to minutes. Decades of studies in secretory cells reveal three exocytosis modes coupled to three endocytosis modes: (a) full-collapse fusion, in which vesicles collapse into the plasma membrane, followed by classical endocytosis involving membrane invagination and vesicle reformation; (b) kiss-and-run, in which the fusion pore opens and closes; and (c) compound exocytosis, which involves exocytosis of giant vesicles formed via vesicle-vesicle fusion, followed by bulk endocytosis that retrieves giant vesicles. Here we review these exo- and endocytosis modes and their roles in regulating quantal size and synaptic strength, generating synaptic plasticity, maintaining exocytosis, and clearing release sites for vesicle replenishment. Furthermore, we highlight recent progress in understanding how vesicle endocytosis is initiated and is thus coupled to exocytosis. The emerging model is that calcium influx via voltage-dependent calcium channels at the calcium microdomain triggers endocytosis and controls endocytosis rate; calmodulin and synaptotagmin are the calcium sensors; and the exocytosis machinery, including SNARE proteins (synaptobrevin, SNAP25, and syntaxin), is needed to coinitiate endocytosis, likely to control the amount of endocytosis.


Keloid tissue analysis discredits a role for myofibroblasts in disease pathogenesis.
Myofibroblasts, renowned for their contractility and extracellular matrix production, are widely considered the key effector cells for nearly all scars resulting from tissue repair processes, ranging from normal scars to extreme fibrosis. For example, it is often assumed that myofibroblasts underpin the characteristics of keloid scars, which are debilitating pathological skin scars lacking effective treatments because of a poor understanding of the disease mechanisms. Here, we present primary and published transcriptional and histological evidence that myofibroblasts are not consistently present in primary keloid lesions, and when alpha-smooth muscle actin (αSMA)-positive cells are detected, they are not greater in number or expressing more αSMA than in normal or hypertrophic scars. In conclusion, keloid scars do not appear to require αSMA-positive myofibroblasts; continuing to consider keloids on a quantitative spectrum with normal or hypertrophic scars, with αSMA serving as a biomarker of disease severity, is hindering advancement of understanding and therapy development.


Differentiation of primate primordial germ cell-like cells following transplantation into the adult gonadal niche.
A major challenge in stem cell differentiation is the availability of bioassays to prove cell types generated in vitro are equivalent to cells in vivo. In the mouse, differentiation of primordial germ cell-like cells (PGCLCs) from pluripotent cells was validated by transplantation, leading to the generation of spermatogenesis and to the birth of offspring. Here we report the use of xenotransplantation (monkey to mouse) and homologous transplantation (monkey to monkey) to validate our in vitro protocol for differentiating male rhesus (r) macaque PGCLCs (rPGCLCs) from induced pluripotent stem cells (riPSCs). Specifically, transplantation of aggregates containing rPGCLCs into mouse and nonhuman primate testicles overcomes a major bottleneck in rPGCLC differentiation. These findings suggest that immature rPGCLCs once transplanted into an adult gonadal niche commit to differentiate towards late rPGCs that initiate epigenetic reprogramming but do not complete the conversion into ENO2-positive spermatogonia.


Lymphangiogenesis in development and disease.
Lymphatic vessels are important for the maintenance of normal tissue fluid balance, for immune surveillance and adsorption of digested fats. In spite of their important functions in physiological as well as in various pathological conditions, including tumour metastasis, lymphoedema and inflammation, the lymphatic vessels have not received as much attention as the blood vessels, and the mechanisms regulating their development and growth have been poorly understood. However, recent studies using mouse genetic tools and primary lymphatic endothelial cell cultures have greatly increased our understanding of how the lymphatic endothelial cells differentiate, how lymphatic vessel growth is regulated and how the remodelling of the lymphatic vasculature into a functional vessel network consisting of capillaries and collecting vessels occurs. Furthermore, these studies have also provided mechanistic insights into the processes involved in pathological lymphangiogenesis.


Cholecystokinin receptors in the brain: characterization and distribution.
Specific cholecystokinin binding sites in particulate fractions of rat brain were measured with iodine 125-labeled Bolton-Hunter cholecystokinin, a cholecystokinin analog that has full biological activity. Binding was detected in brain regions known to contain immunoreactive cholecystokinin. Binding was saturable, reversible, of high affinity (dissociation constant, 1.7 x 10(-9) M), and was inhibited by cholecystokinin analogs but not by unrelated hormones.


Prediction of movement to surgical stimulation by the pupillary dilatation reflex amplitude evoked by a standardized noxious test.
Individual assessment of the amplitude of a physiologic reflex evoked by a standardized noxious test (SNT) before surgical stimulation has been suggested to predict movement upon the forthcoming surgical stimulation. This study aimed to compare the ability of pupillary dilatation reflex amplitude (PDRA) evoked by an SNT and estimated remifentanil effect-site concentration (Ce) to predict movement upon surgical stimulation. Eighty female patients were anesthetized for vacuum aspiration with propofol (Ce 4 μg/ml) and remifentanil. Remifentanil Ce was randomized to 0, 1, 3, or 5 ng/ml. SNT was a 60-mA, 5-s, 100-Hz tetanus applied on median nerve before cervix dilatation. PDRA was calculated as the difference in pupillary diameter after and before SNT. Movement upon cervix dilatation was recorded by an independent observer. Ability of PDRA and estimated remifentanil Ce to discriminate movers from non-movers during cervix dilatation was measured as the area under the receiver operating characteristics curve. Twenty-one of the 76 patients analyzed moved during cervix dilatation. Mean PDRA (±1 SD) evoked by SNT was 2.0 ± 1.2 mm in movers and 0.6 ± 0.7 in non-movers (P < 0.0001). Remifentanil Ce was 0.2 ± 0.4 ng/ml in movers and 3.0 ± 1.7 in non-movers (P < 0.0001). Area under the receiver operating characteristics curve for PDRA was 0.90 (95% CI, 0.83 to 0.96) and for remifentanil Ce 0.94 (0.89 to 0.98), without any significant difference between the two areas. PDRA evoked by an SNT is as accurate as the estimated remifentanil Ce to predict movement upon cervix dilatation. PDRA could be valuable when estimated opioid Ce is not available or reliable.


In vivo requirement of TGF-beta/GDNF cooperativity in mouse development: focus on the neurotrophic hypothesis.
Neurotrophic factors are well-recognized extracellular signaling molecules that regulate neuron development including neurite growth, survival and maturation of neuronal phenotypes in the central and peripheral nervous system. Previous studies have suggested that TGF-beta plays a key role in the regulation of neuron survival and death and potentiates the neurotrophic activity of several neurotrophic factors, most strikingly of GDNF. To test the physiological relevance of this finding, TGF-beta2/GDNF double mutant (d-ko) mice were generated. Double mutant mice die at birth like single mutants due to kidney agenesis (GDNF-/-) and congential cyanosis (TGF-beta2-/-), respectively. To test for the in vivo relevance of TGF-beta2/GDNF cooperativity to regulate neuron survival, mesencephalic dopaminergic neurons, lumbar motoneurons, as well as neurons of the lumbar dorsal root ganglion and the superior cervical ganglion were investigated. No loss of mesencephalic dopaminergic neurons was observed in double mutant mice at E18.5. A partial reduction in neuron numbers was observed in lumbar motoneurons, sensory and sympathetic neurons in GDNF single mutants, which was further reduced in TGF-beta2/GDNF double mutant mice at E18.5. However, TGF-beta2 single mutant mice showed no loss of neurons. These data point towards a cooperative role of TGF-beta2 and GDNF with regard to promotion of survival within the peripheral motor and sensory systems investigated.


Sclerosing peritonitis, a rare complication after intestinal transplant. Report of one case successfully treated with adjustment of immunosuppression.
Sclerosing peritonitis is a complication described in different clinical situations, such as patients that underwent prolonged peritoneal dialysis or renal transplantation with previous history of peritoneal dialysis. The origin of this entity is unclear so far and it is believed that several mechanisms may contribute to its development. The hallmark of sclerosing peritonitis is the continuous accumulation of fibrocollagenous deposits in the intestinal wall and mesenteries causing progressive adhesion of the intestinal loops and mesenteric retraction resulting in intestinal obstruction. Also, it has been described as a rare complication after intestinal transplant that might lead to graft failure. In this report, we describe a case of sclerosing peritonitis after intestinal transplantation that was successfully treated with modifications in the immunosuppressive regime allowing restitution of gastrointestinal transit and intestinal autonomy.


Evaluation on inflammatory states of peripheral veins connected with cannulation /
Peripheral veins cannulation is one of the most frequently executed medical procedures. Primary blood infections which might be the result of peripheral veins cannulation are serious issue that medical care centers all over the world must consider. Medical personnel can choose the area of cannulation thanks to the availability of peripheral veins. Duration of cannulation longer than 72 – 96 hours can cause peripheral veins inflammation. Assay of observation sheets of peripheral cannulation. The study included 14926 patients, who were hospitalized in 2016 in the Municipal Hospital in Zabrze. Observations of each cannulation were recorded in observation sheets for peripheral veins cannulation. Patients age median and duration of cannulation median were calculated basing on 782 observation sheets with registered inflammation of the area of cannulation. Right forearm was area where the most (19%) of vein inflammation occurred. First level of veins inflammation according to criteria of veins inflammation intensity evaluation was the most frequent (90,7%). Duration of venous cannulation median indicated that most of the catheters were removed on the second day. When veins inflammation was diagnosed, the most frequent action was to remove the catheter. During 12 hours 697 observations of cannulation were proceeded. 548 observations of intravenous cannulation were proceeded once for 24 hours. Right forearm was area where the most of cannulations occurred. Frequent observation of the area of peripheral venous cannulation allows quick inflammation symptoms detection.


Fibroblast growth factor-2 bound to specific dermal fibroblast-derived extracellular vesicles is protected from degradation.
Fibroblast growth factor-2 (FGF2) has multiple roles in cutaneous wound healing but its natural low stability prevents the development of its use in skin repair therapies. Here we show that FGF2 binds the outer surface of dermal fibroblast (DF)-derived extracellular vesicles (EVs) and this association protects FGF2 from fast degradation. EVs isolated from DF cultured in the presence of FGF2 harbor FGF2 on their surface and FGF2 can bind purified EVs in absence of cells. Remarkably, FGF2 binding to EVs is restricted to a specific subpopulation of EVs, which do not express CD63 and CD81 markers. Treatment of DF with FGF2-EVs activated ERK and STAT signaling pathways and increased cell proliferation and migration. Local injection of FGF2-EVs improved wound healing in mice. We further demonstrated that binding to EVs protects FGF2 from both thermal and proteolytic degradation, thus maintaining FGF2 function. This suggests that EVs protect soluble factors from degradation and increase their stability and half-life. These results reveal a novel aspect of EV function and suggest EVs as a potential tool for delivering FGF2 in skin healing therapies.


Protective effects of neem (Azadirachta indica A. Juss.) leaf extract against cigarette smoke- and lipopolysaccharide-induced pulmonary inflammation.
Neem (Azadirachta indica A. Juss.) leaf has been reported to exert anti-inflammatory, antibacterial and antioxidant effects. The purpose of this study was to investigate the protective effects of neem leaf extract (NLE) against cigarette smoke (CS)- and lipopolysaccharide (LPS)-induced pulmonary inflammation. Treatment with NLE significantly attenuated the infiltration of inflammatory cells, such as neutrophils and macrophages in bronchoalveolar lavage fluid (BALF). NLE also reduced the production of reactive oxygen species and the activity of neutrophil elastase in BALF. Moreover, NLE attenuated the release of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in BALF. NLE inhibited the recruitment of inflammatory cells and the expression of monocyte chemoattractant protein-1 (MCP-1) in the lungs of mice with CS- and LPS-induced pulmonary inflammation. NLE also decreased the expression of inducible nitric oxide synthase (iNOS) in the lungs of the mice CS- and LPS-induced pulmonary inflammation. Furthermore, treatment with NLE significantly attenuated the activation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) in the lungs mice exposed to CS and LPS. NLE also inhibited the phosphorylation of nuclear factor (NF)-κB and inhibitor of NF-κB (IκB) in the lungs of mice expose to CS and LPS. These findings thus suggest that NLE has potential for use in the treatment of chronic obstructive pulmonary disease.


Economic, social, and environmental evaluation of energy development in the Eagle Ford shale play.
This research investigates the relation between water, energy, and transportation systems, using the growing hydraulic fracturing activity in the Eagle Ford shale play region of southwest Texas in which the local water systems and road infrastructure were not designed for the frequent transport of water into the production site and of produced gas and oil from the site as are often required for hydraulic fracturing. The research: 1) quantifies the interconnections between water, energy, and transportation systems specific to the Eagle Ford shale region; 2) identifies and quantifies the economic, social, and environmental indicators to evaluate scenarios of oil and gas production; and 3) develops a framework for analysis of the economic, societal, and long term sustainability of the sectors and 4) an assessment tool (WET Tool) that estimates several economic indicators: oil and natural gas production, direct and indirect tax revenues, and average wages for each scenario facilitates the holistic assessment of oil and gas production scenarios and their associated trade-offs between them. Additionally, the Tool evaluates these social and environmental indices, (water demand, emissions, water tanker traffic, accidents, road deterioration, and expected average employment times). Scale of production is derived from the price of oil and gas; government revenues from production fluctuations in relation to rise and fall of the oil and gas market prices. While the economic benefits are straightforward, the social costs of shale development (water consumption, carbon emissions, and transportation/infrastructure factors), are difficult to quantify. The tool quantifies and assesses potential scenario outcomes, providing an aid to decision makers in the public and private sectors that allows increased understanding of the implications of each scenario for each sector by summarizing projected outcomes to allow evaluation of the scenarios and comparison of choices and facilitate the essential dialogue between these sectors.


In vitro evaluation of Eugenia dysenterica in primary culture of human gingival fibroblast cells.
Eugenia dysenterica is a Brazilian tree investigated for its properties and bioactive compounds, which are believed to have both pharmacological and phytochemical therapeutic effects. The leaves of this tree contain tannins, flavonoids, terpenes, and saponins, with reportedly beneficial effects to the human body. Despite these therapeutic applications, its effects have never been tested on oral tissues. Therefore, the aim of the present study was to evaluate the cytotoxic and antioxidant effects and the anti-inflammatory and repair properties of the acetone fraction of E. dysenterica on primary culture of human gingival fibroblasts and on the immortalized murine macrophage cell line (RAW 264.7). For this purpose, a metabolic activity assay, a wound healing assay, a nitric oxide assay, and RT-qPCR were performed. The assays revealed a cytoprotective effect of this plant, suggested by the increase in the expression of SOD1 and NRF2. An antioxidant potential effect was observed in the DPPH• assay. However, the fraction of E. dysenterica did not show anti-inflammatory activity. In conclusion, Eugenia dysenterica may promote cytoprotection when associated with chlorhexidine digluconate because of its antioxidant effect. However, additional studies are necessary on other human dental tissues using other parts of the plant in order to develop a possible mouthwash to assist patients with oral disorders.


Nonstandard Work Schedules, Perceived Family Well-Being, and Daily Stressors.
Data from two studies assessed the effects of nonstandard work schedules on perceived family well-being and daily stressors. Study 1, using a sample of employed, married adults aged 25 - 74 (n = 1,166) from the National Survey of Midlife in the United States, showed that night work was associated with perceptions of greater marital instability, negative family-work, and work-family spillover than weekend or daytime work. In Study 2, with a subsample of adults (n = 458) who participated in the National Study of Daily Experiences, weekend workers reported more daily work stressors than weekday workers. Several sociodemographic variables were tested as moderators. Both studies demonstrated that nonstandard work schedules place a strain on working, married adults at the global and daily level.


To What Extent Do Patient Preferences Differ From General Population Preferences?
In some countries including Germany, value sets based on general population preferences are not acceptable for decision-makers in healthcare because the impact of the reference group-general population versus patients-on utility decrements is questioned. The objective of this study was to explore potential differences in patient versus general population health preferences and a way of combining both preferences in economic evaluation. EQ-5D-5L general population preferences were available from national value sets in Germany and Spain. Patient preferences were obtained by conducting discrete choice experiments with patients with rheumatism and patients with diabetes mellitus in Germany and Spain using an online panel. The econometric approach was based on the conditional logit framework. Latent values were anchored using the national value sets. A total of 1700 patients (Germany, n = 937; Spain, n = 763) were included in the analysis. In both countries, patients gave more importance to mobility, self-care, or usual activities and less importance to pain/discomfort and anxiety/depression than the general population. The size of these differences was larger in Germany than in Spain. In Germany, preferences reported by both patient groups were more similar than in Spain. Patient preferences differ from preferences derived from the general population. In contrast to the general population, patients gave more importance to the functional dimensions than to symptoms in both countries. The extent of the differences depends on the disease and the country. For countries preferring patient preferences, a possible way of incorporating the patient perspective in health state valuation was suggested and needs to be further explored.


The dopaminergic stabilizer pridopidine increases neuronal activity of pyramidal neurons in the prefrontal cortex.
The dopaminergic stabilizer pridopidine demonstrates state-dependent effects on locomotor activity, counteracting both hypo- and hyperactivity in rats. Pridopidine has been shown to display both functional dopamine D2 receptor antagonist properties and increase in biomarkers associated with NMDA-mediated glutamate transmission in the frontal cortex. To further characterise the effects of pridopidine on prefrontal cortex (PFC) neurons, a series of in vivo electrophysiological studies were performed in urethane-anaesthetised rats. Pridopidine, administered at doses from 10 to 60 mg/kg (i.v.), dose dependently increased pyramidal cell firing in the majority of the neurons tested. Pridopidine induced a significant increase of 162 % in mean firing activity of PFC neurons, versus initial basal firing activity as the cumulative dose of 30 mg/kg, i.v., was administered. This enhancement of activity was due to increased firing frequency of already spontaneously active neurons, rather than an increase in population activity. The increase was partially reversed or prevented by a sub-threshold dose of the dopamine D1 receptor antagonist SCH23390 (0.5 mg/kg, i.v.). Microiontophoretic application of pridopidine had only moderate activating effects. The selective dopamine D1 receptor agonist A-68930 also had limited effects when administered by microiontophoretic application, but exerted a dose dependent (0.2-3 mg/kg, i.v.) activation of firing in the majority of neurons tested (10/16). However, inhibition of firing by systemic administration of A-68930 was also observed in a subgroup of neurons (6/16). Both activation and inhibition of firing induced by systemic administration of A-68930 were reversed by the systemic administration of SCH23390. The present data suggests that pridopidine enhances pyramidal cell firing via an indirect dopamine D1 receptor-mediated mechanism. These effects of pridopidine may serve to strengthen the cortico-striatal communication and to improve motor control in Huntington's disease for which pridopidine is currently in development.


Effect of buffer infusion during acute respiratory acidosis.
We previously reported that acute respiratory acidosis (ARA) did not stimulate inner medullary collecting duct (IMCD) acidification. It was possible that the failure to find enhanced IMCD acidification was a function of insufficient buffer delivery. To answer this question we studied IMCD acidification in rats with ARA during the infusion of the buffer creatinine. We employed the microcatheterization technique to directly measure pH and PCO2 with glass membrane electrodes and also obtained fluid samples for the measurement of titratable acid and ammonium. Arterial pH was 7.19 +/- 0.01 and PCO2 was 93 +/- 2 mmHg. The IMCD data were analyzed as a function of IMCD length (approximately 6 mm). Equilibrium pH decreased from 5.99 +/- 0.05 to 5.58 +/- 0.02 and PCO2 increased from 71 +/- 11 to 132 +/- 6 mmHg between origin and tip. Bicarbonate delivery decreased from 111 +/- 14 to 38 +/- 2 nmol/min; titratable acid increased from 867 +/- 87 to 1,625 +/- 61 nmol/min, but ammonium delivery did not change along the duct. Thus, estimated net acid increased from 1,772 +/- 155 to 2,709 +/- 88 nmol/min. We conclude that during the presence of increased buffer delivery to the IMCD, rats with ARA markedly increased proton secretion along the terminal nephron.


Conformation of myosin interdomain interactions during contraction: deductions from proteins in solution.
Myosin subfragment 1 (S1) is the ATP catalyzing motor protein in muscle. It consists of three domains that catalyze ATP and bind actin (catalytic), conduct energy transduction (converter), and transport the load (lever arm). These domains interface in two places identified as interface I, containing the reactive thiol (SH1) and ATP-sensitive tryptophan (Trp510), and interface II, containing the reactive lysine residue (RLR). Two crystal structures of S1 were extrapolated to working "in solution" or oriented "in tissue" forms, using structure-sensitive optical spectroscopic signals from extrinsic probes located in the interfaces. Observed signals included circular dichroism (CD) and absorption originating from S1 in solution in the presence and absence of actin and fluorescence polarization from cross-bridges in muscle fibers. Theoretical signals were calculated from S1 crystal structure models perturbed with lever arm movement from swiveling at three conserved glycines, 699, 703, and 710 (chicken skeletal myosin numbering). Structures giving the best agreement between the computed and observed signals were selected as the representative forms. Both interfaces undergo dramatic conformational change during ATPase and force development. Changes at interface I suggest the molecular basis for the collisional quenching sensitivity of Trp510 to nucleotide binding. The probe conformation at SH1 suggests how it alters S1 ATPases. At interface II, the spatial relationship of the lever arm and the extrinsic probe at RLR suggests how the probe alters S1 ATPases and that it should inhibit lever arm movement during the power stroke. The latter possibility, if true, establishes a part of the corridor through which the lever arm swings during the power stroke. Global structural changes in actomyosin are discussed in the accompanying paper [Burghardt et al. (2001) Biochemistry 40, 4821-4833].


[Evaluation of renal damage using urinary ATP analysis].
It is reported that urinary ATP concentration analysis is useful for determining urinary tract infection and renal damage caused by drugs. By means of the firefly luciferin-luciferase method, we determined the reference value of urinary free ATP and evaluated the effects of urine sediments and conditions of storage. The reference value was established as 1.77 x 10(-10) to approximately 7.70 x 10(-9)M using urine samples obtained from 63 outpatients who seemed to have no renal disease. There was no significant difference in ATP concentration between 33 males and 30 females. No significant changes were observed in 11 healthy volunteers during a 1-year period. Within-run reproducibility of ATP was satisfying (8.28% and 11.4% of coefficient value in low and high concentration samples, respectively). ATP concentration was significantly decreased after centrifugation (p < 0.05) and after filtration (p < 0.01). The amounts of the red blood cells (RBC) and white blood cells (WBC) in samples whose ATP concentration was decreased after centrifugation or filtration were significantly higher than those in samples whose concentration did not decrease (p < 0.05). Urine containing many RBCs and/or WBCs might show an artificially higher ATP concentration if no preparations has been performed. There were significant positive correlations between the ATP concentrations before and after refrigeration, but no correlations before and after freezing. It is concluded that the reference value of urinary free ATP concentration was 1.77 x 10(-10) to approximately 7.70 x 10(-9) M and that care is required in the estimation of urinary ATP concentrations in samples containing many sediments, especially with WBC and RBC.


Epilobium angustifolium extract demonstrates multiple effects on dermal fibroblasts in vitro and skin photo-protection in vivo.
Stress-induced fibroblast senescence is thought to contribute to skin aging. Ultraviolet light (UV) radiation is the most potent environmental risk factor in these processes. An Epilobium angustifolium (EA) extract was evaluated for its capacity to reverse the senescent response of normal human dermal fibroblasts (NHDF) in vitro and to exhibit skin photo-protection in vivo. The HPLC-UV-MS analysis of the EA preparation identified three major polyphenol groups: tannins (oenothein B), phenolic acids (gallic and chlorogenic acids) and flavonoids. EA extract increased the cell viability of senescent NHDF induced by serum deprivation. It diminished connective tissue growth factor and fibronectin gene expressions in senescent NHDF. Down-regulation of the UV-induced release of both matrix metalloproteinase-1 and -3 and the tissue inhibitor of matrix metalloproteinases-1 and -2, and also down-regulation of the gene expression of hyaluronidase 2 were observed in repeatedly UV-irradiated NHDF after EA extract treatment. Interestingly, EA extract diminished the down-regulation of sirtuin 1 dampened by UV-irradiation. The application of EA extract using a sub-irritating dose protected skin against UV-induced erythema formation in vivo. In summary, EA extract diminished stress-induced effects on NHDF, particularly on connective tissue growth factor, fibronectin and matrix metalloproteinases. These results collectively suggest that EA extract may possess anti-aging properties and that the EA polyphenols might account for these benefits.


Newly Identified Members of FGFR1 Splice Variants Engage in Cross-talk with AXL/AKT Axis in Salivary Adenoid Cystic Carcinoma.
Adenoid cystic carcinoma (ACC) is the second most common malignancy of the salivary gland. Although characterized as an indolent tumor, ACC often leads to incurable metastatic disease. Patients with ACC respond poorly to currently available therapeutic drugs and factors contributing to the limited response remain unknown. Determining the role of molecular alterations frequently occurring in ACC may clarify ACC tumorigenesis and advance the development of effective treatment strategies. Applying Splice Expression Variant Analysis and outlier statistics on RNA sequencing of primary ACC tumors and matched normal salivary gland tissues, we identified multiple alternative splicing events (ASE) of genes specific to ACC. In ACC cells and patient-derived xenografts, FGFR1 was a uniquely expressed ASE. Detailed PCR analysis identified three novel, truncated, intracellular domain-lacking FGFR1 variants (FGFR1v). Cloning and expression analysis suggest that the three FGFR1v are cell surface proteins, that expression of FGFR1v augmented pAKT activity, and that cells became more resistant to pharmacologic FGFR1 inhibitor. FGFR1v-induced AKT activation was associated with AXL function, and inhibition of AXL activity in FGFR1v knockdown cells led to enhanced cytotoxicity in ACC. Moreover, cell killing effect was increased by dual inhibition of AXL and FGFR1 in ACC cells. This study demonstrates that these previously undescribed FGFR1v cooperate with AXL and desensitize cells to FGFR1 inhibitor, which supports further investigation into combined FGFR1 and AXL inhibition as an effective ACC therapy.This study identifies several FGFR1 variants that function through the AXL/AKT signaling pathway independent of FGF/FGFR1, desensitizing cells to FGFR1 inhibitor suggestive of a potential resistance mechanism in ACC. SIGNIFICANCE: This study identifies several FGFR1 variants that function through the AXL/AKT signaling pathway independent of FGF/FGFR1, desensitizing cells to FGFR1 inhibitor, suggestive of a potential resistance mechanism in ACC.


The influence of carbohydrate-electrolyte ingestion on soccer skill performance.
To investigate the effect of ingesting a carbohydrate-electrolyte solution (CHO-E), in subjects with reduced carbohydrate stores, during an intermittent shuttle running test (LIST) on soccer passing (LSPT) and shooting (LSST) performance. Sixteen healthy male university soccer players ingested either a 6.4% CHO-E or placebo (PLA) solution during 90 min of the LIST (5 mL x kg(-1) BM before and 2 mL x kg(-1) BM every 15 min of exercise), in a double-blind, randomized, crossover design, with each trial separated by at least 7 d. On the evening before the main trial (17:00 h), subjects performed the glycogen-reducing cycling exercise (approximately 80 min at 70% VO2max). They were then fed a low-carbohydrate evening meal and reported to the laboratory the following morning after a 10-h fast. Blood was collected at rest and after every 30 min of exercise; skill tests were performed before and after the LIST. The change in mean LSST performance from pre- to post-LIST was better in the CHO-E trial (11 +/- 45 vs -16 +/- 42%; P < 0.01) but not significantly different for the LSPT performance (-1 +/- 10% (CHO-E) vs -6 +/- 13% (PLA), P = 0.13). Sprint performance during the LIST was quicker in the CHO-E trial (2.50 +/- 0.13 vs 2.53 +/- 0.13 s, P < 0.01). Plasma glucose was higher in the CHO-E trial after 90 min of exercise (5.2 +/- 0.3 vs 3.9 +/- 0.4 mM, P < 0.01). Ingestion of a carbohydrate-electrolyte solution during exercise enabled subjects with compromised glycogen stores to better maintain skill and sprint performance than when ingesting fluid alone.


Multisensory processing in spatial orientation: an inverse probabilistic approach.
Most evidence that the brain uses Bayesian inference to integrate noisy sensory signals optimally has been obtained by showing that the noise levels in each modality separately can predict performance in combined conditions. Such a forward approach is difficult to implement when the various signals cannot be measured in isolation, as in spatial orientation, which involves the processing of visual, somatosensory, and vestibular cues. Instead, we applied an inverse probabilistic approach, based on optimal observer theory. Our goal was to investigate whether the perceptual differences found when probing two different states--body-in-space and head-in-space orientation--can be reconciled by a shared scheme using all available sensory signals. Using a psychometric approach, seven human subjects were tested on two orientation estimates at tilts < 120°: perception of body tilt [subjective body tilt (SBT)] and perception of visual vertical [subjective visual vertical (SVV)]. In all subjects, the SBT was more accurate than the SVV, which showed substantial systematic errors for tilt angles beyond 60°. Variability increased with tilt angle in both tasks, but was consistently lower in the SVV. The sensory integration model fitted both datasets very nicely. A further experiment, in which supine subjects judged their head orientation relative to the body, independently confirmed the predicted head-on-body noise by the model. Model predictions based on the derived noise properties from the various modalities were also consistent with previously published deficits in vestibular and somatosensory patients. We conclude that Bayesian computations can account for the typical differences in spatial orientation judgments associated with different task requirements.


Tuberculosis and HIV control in sub-Saharan African prisons: "thinking outside the prison cell".
Tuberculosis is one of the fastest-growing epidemics in prison populations in sub-Saharan Africa (SSA), constituting a threat to both inmates and the wider community. Various factors have contributed to the breakdown of tuberculosis control in prison facilities in SSA, including slow and insensitive diagnostics, failing prison infrastructure, inadequate funding, and weak prevention and treatment interventions for human immunodeficiency virus (HIV). In this article, we describe the challenges inherent in current approaches to tuberculosis control in prisons and consider the alternatives. We argue that although improved implementation of conventional tuberculosis control activities is necessary, considerable investment in a broader range of public health interventions, including infrastructure and staffing upgrades, cutting-edge tuberculosis diagnostics, and combination prevention for HIV, will be equally critical. This combination response to tuberculosis in prisons will be essential for tackling existing and nascent prison tuberculosis epidemics and will require high-level political support and financing.


Effect of poly(amido)amine (PAMAM) G4 dendrimer on heart and liver mitochondria in an animal model of diabetes.
Diabetes-induced injury related to hyperglycaemia is associated with impaired function of mitochondria. Regardless of their cytotoxicity, PAMAM [poly(amido)amine] G4 dendrimers lower plasma glucose and suppress long-term markers of diabetic hyperglycaemia in experimental diabetes. In the present study, we aimed at verifying whether such modulatory effects of PAMAM G4 (0.5 micromol/kg of body weight daily for 60 days) may contribute to improved respiration in heart and liver mitochondria from streptozotocin-diabetic rats. PAMAM G4 alleviated long-term markers of hyperglycaemia and reduced blood and tissue lipophilic antioxidants in diabetic animals, but did not restore mitochondrial function. In hearts, but not livers, dendrimers further reduced respiratory function and oxidative phosphorylation. Thus ameliorating effects of PAMAM G4 on glycation and glycoxidation in experimental diabetes are not sufficient to restore the impaired mitochondrial function in diabetes.


Assessment of Residual Cancer Burden and Event-Free Survival in Neoadjuvant Treatment for High-risk Breast Cancer: An Analysis of Data From the I-SPY2 Randomized Clinical Trial.
Residual cancer burden (RCB) distributions may improve the interpretation of efficacy in neoadjuvant breast cancer trials. To compare RCB distributions between randomized control and investigational treatments within subtypes of breast cancer and explore the relationship with survival. The I-SPY2 is a multicenter, platform adaptive, randomized clinical trial in the US that compares, by subtype, investigational agents in combination with chemotherapy vs chemotherapy alone in adult women with stage 2/3 breast cancer at high risk of early recurrence. Investigational treatments graduated in a prespecified subtype if there was 85% or greater predicted probability of higher rate of pathologic complete response (pCR) in a confirmatory, 300-patient, 1:1 randomized, neoadjuvant trial in that subtype. Evaluation of a secondary end point was reported from the 10 investigational agents tested in the I-SPY2 trial from March 200 through 2016, and analyzed as of September 9, 2020. The analysis plan included modeling of RCB within subtypes defined by hormone receptor (HR) and ERBB2 status and compared control treatments with investigational treatments that graduated and those that did not graduate. Neoadjuvant paclitaxel plus/minus 1 of several investigational agents for 12 weeks, then 12 weeks of cyclophosphamide/doxorubicin chemotherapy followed by surgery. Residual cancer burden (pathological measure of residual disease) and event-free survival (EFS). A total of 938 women (mean [SD] age, 49 [11] years; 66 [7%] Asian, 103 [11%] Black, and 750 [80%] White individuals) from the first 10 investigational agents were included, with a median follow-up of 52 months (IQR, 29 months). Event-free survival worsened significantly per unit of RCB in every subtype of breast cancer (HR-positive/ERBB2-negative: hazard ratio [HZR], 1.75; 95% CI, 1.45-2.16; HR-positive/ERBB2-positive: HZR, 1.55; 95% CI, 1.18-2.05; HR-negative/ERBB2-positive: HZR, 2.39; 95% CI, 1.64-3.49; HR-negative/ERBB2-negative: HZR, 1.99; 95% CI, 1.71-2.31). Prognostic information from RCB was similar from treatments that graduated (HZR, 2.00; 95% CI, 1.57-2.55; 254 [27%]), did not graduate (HZR, 1.87; 95% CI, 1.61-2.17; 486 [52%]), or were control (HZR, 1.79; 95% CI, 1.42-2.26; 198 [21%]). Investigational treatments significantly lowered RCB in HR-negative/ERBB2-negative (graduated and nongraduated treatments) and ERBB2-positive subtypes (graduated treatments), with improved EFS (HZR, 0.61; 95% CI, 0.41-0.93) in the exploratory analysis. In this randomized clinical trial, the prognostic significance of RCB was consistent regardless of subtype and treatment. Effective neoadjuvant treatments shifted the distribution of RCB in addition to increasing pCR rate and appeared to improve EFS. Using a standardized quantitative method to measure response advances the interpretation of efficacy. ClinicalTrials.gov Identifier: NCT01042379.


Variability in the response of the cyanobacterium Nodularia spumigena to nitrogen supplementation.
Nodularia spumigena is a filamentous cyanobacterium that is commonly found in brackish water bodies. The species is capable of fixing atmospheric nitrogen in specialized cells termed heterocysts. N. spumigena dominates the annual toxic summer blooms in the Baltic Sea causing environmental and economical problems. We have previously demonstrated that N. spumigena strain AV1 exhibits a different response to the presence of combined nitrogen as compared to model cyanobacteria such as Nostoc PCC7120 and Nostoc punctiforme by uncoupling between nitrogen fixation and heterocyst differentiation.In order to assess whether or not the behaviour of N. spumigena strain AV1 is characteristic of N. spumigena populations, especially in the Baltic Sea, we have investigated the effect of nitrate and ammonium ions on growth, nitrogen fixation activity and presence of heterocysts in eight different Nodularia strains. Our results show that most of the strains retain heterocysts in the absence of nitrogenase activity indicating that uncoupling between nitrogen fixation and heterocyst differentiation is most likely a common behaviour among N. spumigena strains, especially in the Baltic Sea. In addition, there are variations between strains in regard to nitrate uptake.


[Formation of plastic connections of the visual system during ontogenesis of the rabbit].
Analysis of evoked potentials and unit activity in the visual cortical projection area of rabbits revealed a definite succession of forming of interneuronal connections in ontogeny. In early postnatal period, the neuronal reactions were characterized by stable responses with one excitatory phase corresponding to initially negative surface evoked potential. Similarity of reactions of neurones situated in the same vertical column was observed and explained by functioning of a system of rigid connections of the thalamic relay nuclei afferents with cortical pyramidal neurones. Beginning from the third week of postnatal life of rabbits the neuronal reactions assumed a distinctly expressed phasic character, and variability of responses was seen along the vertical line. The changes revealed correlated with formation of a system of interneurones providing a possibility of plastic neuronal interaction. A study of the influence of preliminary cortical stimulation of the associative areas showed that intercentral cooperation mediated by cortical interneurones providing a systemic analysis of visual information began to form from the third week of postnatal life and reached the definitive level at later stages of development.


Apocrine hidrocystomas of the lids, hypodontia, palmar-plantar hyperkeratosis, and onychodystrophy. A new variant of ectodermal dysplasia.
A fourth case of a newly recognized variant of ectodermal dysplasia is reported. This syndrome is characterized by bilateral apocrine hidrocystomas of the eyelid margins, hypodontia, palmar-plantar hyperkeratosis, and onychodystrophy. To our knowledge, this syndrome, with its striking ocular manifestations, has not been previously documented in the ophthalmic literature.


[Hyperlipidemia and decrease of lipid tolerance as factors of atherogenesis].
In experiments performed on rabbits the influence of acute and chronic alimentary lipid loading on the pathogenic factors of atherosclerosis was examined. A single moderate lipid loading even in normal rabbits was established to induce a transitory development of the proatherogenic changes, which includes appearance of dislipidemia of diabetic type, activation of systemic inflammation and free radical reactions, blood lipoprotein modification. Prolonged keeping of rabbits on the diet enriched with lipids induced the development of complex changes which are typical for insulin resistance with specific lipid and lipoprotein metabolism disturbances, increased plasma atherogenic potential, activation of systemic inflammation and oxidative stress. We also noticed a sharp decrease of lipid tolerance and acute lipid loading was accompanied by more pronounced changes than in normal rabbits. The data show that the excessive consumption of alimentary lipids can induce the development and sharp activation of atherosclerosis pathogenic factors.


A Practical, One-Clinic Visit Protocol for Pharmacokinetic Profile Generation with the ADVATE myPKFiT Dosing Tool in Severe Hemophilia A Subjects.
Standard pharmacokinetic (PK) assessments are demanding for persons with hemophilia A, requiring a 72-hour washout and 5 to 11 timed blood samples. A no-washout, single-clinic visit, sparse sampling population PK (PPK) protocol is an attractive alternative. Here, we compared PK parameters obtained with a traditional washout, 6-sampling time point PPK protocol with a no-washout, single-clinic visit, reverse 2-sampling time point PPK protocol in persons with severe hemophilia A (SHA) receiving ADVATE. A total of 39 inhibitor-negative males with SHA (factor VIII activity [FVIII:C] < 2%) were enrolled in a prospective sequential design PK study. Participants completed a washout, 6-sampling time point PPK protocol as well as a no-washout, reverse 2-sampling time point protocol, with samples taken during a single 3-hour clinic visit 24 hours post home infusion of FVIII and then 3 hours post infusion in clinic. FVIII:C levels were analyzed by one-stage and chromogenic assays; blood group and von Willebrand factor antigen (VWF:Ag) were determined; and PK parameters were analyzed using the ADVATE myPKFiT dosing tool. There was moderate to almost perfect agreement for the PK parameters obtained with the 2- and the 6- point PPK protocols using a one-stage FVIII:C assay and a substantial to almost perfect agreement using a chromogenic FVIII:C assay. Significant associations between specific PK parameters and blood group and VWF:Ag were observed. The no-washout, single-clinic visit, reverse 2-sampling time point PPK protocol can be used in the routine clinical setting since it demonstrates sufficient accuracy compared with the more demanding and less practical washout, 6-sampling time point PPK protocol in persons with SHA receiving ADVATE.


Comparison of medication errors in an American and a British hospital.
Medication errors in a hospital in the United States and a hospital in the United Kingdom were compared. The study was conducted in wards with a high oral-drug-related workload in two large university hospitals. The U.S. hospital was studied in August 1993 and the U.K. hospital in May and June 1993. The U.S. hospital had a typical unit dose drug distribution system, and the U.K. hospital had the ward-based system commonly used in that country, in which a pharmacist visits each ward several times daily and reviews each patient's medication chart. The medication chart is used by the physician to order drugs and obviates the need for transcription of orders. A disguised-observation technique was used to determine frequencies and types of medication errors. Medication errors were identified retrospectively in the U.S. hospital by comparing the observer's notes with the original drug orders made in the patient's chart by the physician. In the U.K. hospital, identification of errors took place concurrently; as doses were administered, they were compared with the orders on the medication chart. In the U.S. and U.K. hospitals, 919 and 2756 opportunities for error were observed, respectively. The medication error rate in the U.S. hospital was 6.9% (95% confidence interval [CI], 5.2% to 8.5%), significantly higher than the 3.0% rate observed in the U.K. hospital (95% CI, 2.4% to 3.7%) (95% CI for the difference, 2.1% to 5.7%). Omitted doses and incorrect doses were the most common types of errors in the U.K. hospital; incorrect doses and unordered doses were the most common types in the U.S. hospital. An American hospital with a unit dose distribution system had a significantly higher medication error rate than a British hospital with a ward-based supply system.