Maternal androgen excess and obesity induce sexually dimorphic anxiety-like behavior in the offspring.
Maternal polycystic ovary syndrome (PCOS), a condition associated with hyperandrogenism, is suggested to increase anxiety-like behavior in the offspring. Because PCOS is closely linked to obesity, we investigated the impact of an adverse hormonal or metabolic maternal environment and offspring obesity on anxiety in the offspring. The obese PCOS phenotype was induced by chronic high-fat-high-sucrose (HFHS) consumption together with prenatal dihydrotestosterone exposure in mouse dams. Anxiety-like behavior was assessed in adult offspring with the elevated-plus maze and open-field tests. The influence of maternal androgens and maternal and offspring diet on genes implicated in anxiety were analyzed in the amygdala and hypothalamus with real-time PCR ( n = 47). Independent of diet, female offspring exposed to maternal androgens were more anxious and displayed up-regulation of adrenoceptor α 1B in the amygdala and up-regulation of hypothalamic corticotropin-releasing hormone ( Crh). By contrast, male offspring exposed to a HFHS maternal diet had increased anxiety-like behavior and showed up-regulation of epigenetic markers in the amygdala and up-regulation of hypothalamic Crh. Overall, there were substantial sex differences in gene expression in the brain. These findings provide novel insight into how maternal androgens and obesity exert sex-specific effects on behavior and gene expression in the offspring of a PCOS mouse model.-Manti, M., Fornes, R., Qi, X., Folmerz, E., Lindén Hirschberg, A., de Castro Barbosa, T., Maliqueo, M., Benrick, A., Stener-Victorin, E. Maternal androgen excess and obesity induce sexually dimorphic anxiety-like behavior in the offspring.


Lack of short-wavelength light during the school day delays dim light melatonin onset (DLMO) in middle school students.
Circadian timing affects sleep onset. Delayed sleep onset can reduce sleep duration in adolescents required to awake early for a fixed school schedule. The absence of short-wavelength ("blue") morning light, which helps entrain the circadian system, can hypothetically delay sleep onset and decrease sleep duration in adolescents. The goal of this study was to investigate whether removal of short-wavelength light during the morning hours delayed the onset of melatonin in young adults. Dim light melatonin onset (DLMO) was measured in eleven 8th-grade students before and after wearing orange glasses, which removed short-wavelength light, for a five-day school week. DLMO was significantly delayed (30 minutes) after the five-day intervention, demonstrating that short-wavelength light exposure during the day can be important for advancing circadian rhythms in students. Lack of short-wavelength light in the morning has been shown to delay the circadian clock in controlled laboratory conditions. The results presented here are the first to show, outside laboratory conditions, that removal of short-wavelength light in the morning hours can delay DLMO in 8th-grade students. These field data, consistent with results from controlled laboratory studies, are directly relevant to lighting practice in schools.


Forest structure in low-diversity tropical forests: a study of Hawaiian wet and dry forests.
The potential influence of diversity on ecosystem structure and function remains a topic of significant debate, especially for tropical forests where diversity can range widely. We used Center for Tropical Forest Science (CTFS) methodology to establish forest dynamics plots in montane wet forest and lowland dry forest on Hawai'i Island. We compared the species diversity, tree density, basal area, biomass, and size class distributions between the two forest types. We then examined these variables across tropical forests within the CTFS network. Consistent with other island forests, the Hawai'i forests were characterized by low species richness and very high relative dominance. The two Hawai'i forests were floristically distinct, yet similar in species richness (15 vs. 21 species) and stem density (3078 vs. 3486/ha). While these forests were selected for their low invasive species cover relative to surrounding forests, both forests averaged 5->50% invasive species cover; ongoing removal will be necessary to reduce or prevent competitive impacts, especially from woody species. The montane wet forest had much larger trees, resulting in eightfold higher basal area and above-ground biomass. Across the CTFS network, the Hawaiian montane wet forest was similar to other tropical forests with respect to diameter distributions, density, and aboveground biomass, while the Hawai'i lowland dry forest was similar in density to tropical forests with much higher diversity. These findings suggest that forest structural variables can be similar across tropical forests independently of species richness. The inclusion of low-diversity Pacific Island forests in the CTFS network provides an ∼80-fold range in species richness (15-1182 species), six-fold variation in mean annual rainfall (835-5272 mm yr(-1)) and 1.8-fold variation in mean annual temperature (16.0-28.4°C). Thus, the Hawaiian forest plots expand the global forest plot network to enable testing of ecological theory for links among species diversity, environmental variation and ecosystem function.


Benefits of physical training in patients with idiopathic or end-stage sarcoidosis-related pulmonary fibrosis: a pilot study.
The natural history of disease in patients with stage IV (fibrotic) sarcoidosis may mirror that of patients with idiopathic pulmonary fibrosis (IPF). Both are bothered by progressive dyspnea, exercise limitation and fatigue. To establish whether patients suffering from pulmonary fibrosis might benefit from a physical training program. Twenty-four eligible patients referred to the out-patient clinic of the ild care expertise team of Hospital Gelderse Vallei, Ede, The Netherlands between November 2012 and November 2013 were included in this observational pilot study of a 12-week physical training program. Outcomes, including exercise capacity, skeletal muscle strength, lung function and fatigue were assessed at two time points: 1) baseline; and 2) after completion of a 12-week physical training program. At baseline, the percentage predicted DLCO, FVC, FEV1 and exercise capacity (assessed by six-minute walking distance (6MWD) or maximal oxygen uptake) was reduced in both groups. After program completion, exercise capacity improved (>10% improvement 6MWD) in 13 subjects (54.2%): 7 with IPF and 6 with sarcoidosis subjects. Other secondary endpoints, including pulmonary function tests and patient-reported outcome measures improved in some subjects. A 12-week physical training program improved or maintained exercise capacity in patients with IPF (despite disease progression) or fibrotic sarcoidosis. The results from this pilot study could be used to design prospective studies aimed at answering lingering questions about exercise training in patients with these progressive, incurable conditions.


A quantitative evaluation of mucosal eosinophils in the pediatric gastrointestinal tract.
Evaluation of the mucosal eosinophil content is important in the interpretation of endoscopic biopsies, as high eosinophil densities might reflect allergic gastrointestinal disease. Reference ranges gleaned from the literature have an upper limit of normal varying from 6 to 20 eosinophils per 400 x high power field. Preliminary data suggest a geographic variation with higher eosinophil counts in the southern United States. We examined intestinal tract mucosa from 44 infants and children who died suddenly and unexpectedly in northeastern Texas. Subjects ranged in age from 3 wks to 17 yrs and were without known gastrointestinal disease. Using formalin-fixed, hematoxylin and eosin-stained 4-microns sections, intramucosal eosinophils were counted in ten consecutive high power fields and the mean eosinophil count determined. Twenty-three subjects (52%) had counts of > 20 eosinophils/high power field from at least one site. There was no correlation with age, sex, season, or cause of death. In a subset of 11 subjects, more extensive sampling showed the cecum and appendix to have the highest concentration of eosinophils and relatively low counts in the stomach and distal large intestine. These observations correlate with our impression that increased numbers of eosinophils, particularly in the proximal colon, are a common feature of otherwise unremarkable pediatric endoscopic biopsies. Efforts to distinguish the proportion of activated eosinophils in B5-fixed mucosal biopsies using the EG2 monoclonal antibody were unsuccessful, as this antibody does not appear specific for activation in B5-fixed tissue. Mucosal eosinophil counts should be interpreted with caution in geographic areas where a high background count is endemic.


The transcriptional repressor NKAP is required for the development of iNKT cells.
Invariant natural killer T cells have a distinct developmental pathway from conventional αβ T cells. Here we demonstrate that the transcriptional repressor NKAP is required for invariant natural killer T cell but not conventional T cell development. In CD4-cre NKAP conditional knockout mice, invariant natural killer T cell development is blocked at the double-positive stage. This cell-intrinsic block is not due to decreased survival or failure to rearrange the invariant Vα14-Jα18 T cell receptor-α chain, but is rescued by overexpression of a rec-Vα14-Jα18 transgene at the double-positive stage, thus defining a role for NKAP in selection into the invariant natural killer T cell lineage. Importantly, deletion of the NKAP-associated protein histone deacetylase 3 causes a similar block in the invariant natural killer T cell development, indicating that NKAP and histone deacetylase 3 functionally interact to control invariant natural killer T cell development.


A small amount of mini-charged dark matter could cool the baryons in the early Universe.
The dynamics of our Universe is strongly influenced by pervasive-albeit elusive-dark matter, with a total mass about five times the mass of all the baryons1,2. Despite this, its origin and composition remain a mystery. All evidence for dark matter relies on its gravitational pull on baryons, and thus such evidence does not require any non-gravitational coupling between baryons and dark matter. Nonetheless, some small coupling would explain the comparable cosmic abundances of dark matter and baryons 3 , as well as solving structure-formation puzzles in the pure cold-dark-matter models 4 . A vast array of observations has been unable to find conclusive evidence for any non-gravitational interactions of baryons with dark matter5-9. Recent observations by the EDGES collaboration, however, suggest that during the cosmic dawn, roughly 200 million years after the Big Bang, the baryonic temperature was half of its expected value 10 . This observation is difficult to reconcile with the standard cosmological model but could be explained if baryons are cooled down by interactions with dark matter, as expected if their interaction rate grows steeply at low velocities 11 . Here we report that if a small fraction-less than one per cent-of the dark matter has a mini-charge, a million times smaller than the charge on the electron, and a mass in the range of 1-100 times the electron mass, then the data 10 from the EDGES experiment can be explained while remaining consistent with all other observations. We also show that the entirety of the dark matter cannot have a mini-charge.


Effects of inhaled nitric oxide compared with aspirin on platelet function in vivo in healthy subjects.
1. Nitric oxide has platelet-stabilizing effects. Inhaled nitric oxide is used to treat pulmonary disorders, and may prolong bleeding times, suggesting that it has effects on haemostasis. We therefore examined if inhaled nitric oxide influences platelet function in vivo in healthy subjects. 2. Platelet aggregability (filtragometry ex vivo, which reflects aggregability in vivo), bleeding time and platelet secretion products and cGMP in plasma were studied during inhalation of two different doses of nitric oxide (30 and 80 p.p.m.; 15 min at each dose level; n = 19) and during prolonged (55 min; n = 18) inhalation of 30 p.p.m. nitric oxide. For comparison, studies were also performed before and after ingestion of 640 mg aspirin in 13 of the healthy subjects. 3. Plasma cGMP increased dose dependently during nitric oxide inhalation, suggesting guanylate cyclase activation in vivo. Platelet aggregability was, however, little affected and platelet secretion was not attenuated by nitric oxide inhalation. Bleeding time tended to increase (by 16-33%), but was significantly increased only after prolonged inhalation of nitric oxide at 30 p.p.m. 4. Aspirin (640 mg orally) caused pronounced and significant prolongations of filtragometry readings and bleeding time. Thus, the methods used were able to reveal platelet stabilization. 5. We conclude that nitric oxide inhalation causes only mild, if any, attenuation of platelet function in healthy subjects with a normal endogenous nitric oxide production. The effects may be different in disease states.


Aerobic biodegradation of N-nitrosodimethylamine (NDMA) by axenic bacterial strains.
The water contaminant N-nitrosodimethylamine (NDMA) is a probable human carcinogen whose appearance in the environment is related to the release of rocket fuel and to chlorine-based disinfection of water and wastewater. Although this compound has been shown to be biodegradable, there is minimal information about the organisms capable of this degradation, and little is understood of the mechanisms or biochemistry involved. This study shows that bacteria expressing monooxygenase enzymes functionally similar to those demonstrated to degrade NDMA in eukaryotes have the capability to degrade NDMA. Specifically, induction of the soluble methane monooxygenase (sMMO) expressed by Methylosinus trichosporium OB3b, the propane monooxygenase (PMO) enzyme of Mycobacterium vaccae JOB-5, and the toluene 4-monooxygenases found in Ralstonia pickettii PKO1 and Pseudomonas mendocina KR1 resulted in NDMA degradation by these strains. In each of these cases, brief exposure to acetylene gas, a suicide substrate for certain monooxygenases, inhibited the degradation of NDMA. Further, Escherichia coli TG1/pBS(Kan) containing recombinant plasmids derived from the toluene monooxygenases found in strains PKO1 and KR1 mimicked the behavior of the parent strains. In contrast, M. trichosporium OB3b expressing the particulate form of MMO, Burkholderia cepacia G4 expressing the toluene 2-monooxygenase, and Pseudomonas putida mt-2 expressing the toluene sidechain monooxygenase were not capable of NDMA degradation. In addition, bacteria expressing aromatic dioxygenases were not capable of NDMA degradation. Finally, Rhodococcus sp. RR1 exhibited the ability to degrade NDMA by an unidentified, constitutively expressed enzyme that, unlike the confirmed monooxygenases, was not inhibited by acetylene exposure.


Stereotactic Ablative Radiotherapy for the Management of Spinal Metastases: A Review.
Rising cancer incidence combined with improvements in systemic and local therapies extending life expectancy are translating into more patients with spinal metastases. This makes the multidisciplinary management of spinal metastases and development of new therapies increasingly important. Spinal metastases may cause significant pain and reduced quality of life and lead to permanent neurological disability if compression of the spinal cord and/or nerve root occurs. Until recently, treatments for spinal metastases were not optimal and provided temporary local control and pain relief. Spinal stereotactic ablative radiotherapy (SABR) is an effective approach associated with an improved therapeutic ratio, with evolving clinical application. To review the literature of spinal SABR for spinal metastases, discuss a multidisciplinary approach to appropriate patient selection and technical considerations, and summarize current efforts to combine spinal SABR with systemic therapies. The MEDLINE database was searched to identify articles reporting on spinal SABR to September 30, 2018. Articles including clinical trials, prospective and retrospective studies, systematic reviews, and consensus recommendations were selected for relevance to multidisciplinary management of spinal metastases. Fifty-nine unique publications with 5655 patients who underwent SABR for spinal metastases were included. Four comprehensive frameworks for patient selection were discussed. Spinal SABR was associated with 1-year local control rates of approximately 80% to 90% in the de novo setting, greater than 80% in the postoperative setting, and greater than 65% in the reirradiation setting. The most commonly discussed adverse effect was development of a vertebral compression fracture with variable rates, most commonly reported as approximately 10% to 15%. High-level data on the combination of SABR with modern therapies are still lacking. At present, 19 clinical trials are ongoing, mainly focusing on combined modality therapies, radiotherapy prescription dose, and oligometastic disease. These findings suggest that spinal SABR may be an effective treatment option for well-selected patients with spinal metastases, achieving high rates of local tumor control with moderate rates of adverse effects. Optimal management should include review by a multidisciplinary care team.


Extracellular DNA traps are associated with the pathogenesis of TRALI in humans and mice.
Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related death. The biologic processes contributing to TRALI are poorly understood. All blood products can cause TRALI, and no specific treatment is available. A "2-event model" has been proposed as the trigger. The first event may include surgery, trauma, or infection; the second involves the transfusion of antileukocyte antibodies or bioactive lipids within the blood product. Together, these events induce neutrophil activation in the lungs, causing endothelial damage and capillary leakage. Neutrophils, in response to pathogens or under stress, can release their chromatin coated with granule contents, thus forming neutrophil extracellular traps (NETs). Although protective against infection, these NETs are injurious to tissue. Here we show that NET biomarkers are present in TRALI patients' blood and that NETs are produced in vitro by primed human neutrophils when challenged with anti-HNA-3a antibodies previously implicated in TRALI. NETs are found in alveoli of mice experiencing antibody-mediated TRALI. DNase 1 inhalation prevents their alveolar accumulation and improves arterial oxygen saturation even when administered 90 minutes after TRALI onset. We suggest that NETs form in the lungs during TRALI, contribute to the disease process, and thus could be targeted to prevent or treat TRALI.


Impact of HLA-DPB1 haplotypes on outcome of 10/10 matched unrelated hematopoietic stem cell donor transplants depends on MHC-linked microsatellite polymorphisms.
Hematopoietic stem cell transplantation (HSCT) with HLA-A, -B, -C, -DRB1, -DQB1 allele matched (10 of 10) unrelated donors is still associated with a significant rate of posttransplantation complications. In order to disclose additional immunogenetic factors, we analyzed the impact of HLA-DPB1 disparities and major histocompatibility complex (MHC)-resident microsatellite polymorphisms in 246 HLA 10 of 10 matched HSCT patients. First we showed that patients with more frequent/conserved HLA haplotypes had a higher 5-year survival (55% ± 18% versus 39% ± 18%, P = .021). In addition, DPB1 incompatibilities and 3 microsatellite alleles were associated with outcome. In a Cox regression model adjusting for European Blood and Marrow Transplant (EBMT) risk score, T cell depletion, and year of treatment, HSCT with a tumor necrosis factor d (TNFd) 4/d5-positive donor was associated with increased mortality (hazard ratio [HR] = 2.03; confidence interval [CI] 1.25-3.31; P = .004), whereas the D6S510-184 allele was protective (HR = 0.44; CI 0.22-0.87; P = .018). The 2 MHC-linked genetic donor factors, DPB1 mismatch (MM), and TNFd4/d5-positivity, acted in synergy with the EBMT risk score with an always lower survival (HR = 2.97; CI 1.27-6.92; P = .012). These data show that multiple MHC-linked genetic donor factors impact on outcome after unrelated donor HSCT. Their additive and potentially divergent effects could explain previous discrepant results, particularly with respect to the role of HLA-DPB1 disparities. We conclude that HLA-DPB1 typing combined with a simple TNFd microsatellite genotyping assay may significantly help in pretransplantation risk assessment for graft-versus-host disease and mortality, particularly for patients with several potential 10 of 10 matched donors.


MicroRNAs: crucial regulators of placental development.
MicroRNAs (miRNAs) are small non-coding single-stranded RNAs that are integral to a wide range of cellular processes mainly through the regulation of translation and mRNA stability of their target genes. The placenta is a transient organ that exists throughout gestation in mammals, facilitating nutrient and gas exchange and waste removal between the mother and the fetus. miRNAs are expressed in the placenta, and many studies have shown that miRNAs play an important role in regulating trophoblast differentiation, migration, invasion, proliferation, apoptosis, vasculogenesis/angiogenesis and cellular metabolism. In this review, we provide a brief overview of canonical and non-canonical pathways of miRNA biogenesis and mechanisms of miRNA actions. We highlight the current knowledge of the role of miRNAs in placental development. Finally, we point out several limitations of the current research and suggest future directions.


A multi-country outbreak of Salmonella Newport gastroenteritis in Europe associated with watermelon from Brazil, confirmed by whole genome sequencing: October 2011 to January 2012.
In November 2011, the presence of Salmonella Newport in a ready-to-eat watermelon slice was confirmed as part of a local food survey in England. In late December 2011, cases of S. Newport were reported in England, Wales, Northern Ireland, Scotland, Ireland and Germany. During the outbreak, 63 confirmed cases of S. Newport were reported across all six countries with isolates indistinguishable by pulsed-field gel electrophoresis from the watermelon isolate.A subset of outbreak isolates were whole-genome sequenced and were identical to, or one single nucleotide polymorphism different from the watermelon isolate.In total, 46 confirmed cases were interviewed of which 27 reported watermelon consumption. Further investigations confirmed the outbreak was linked to the consumption of watermelon imported from Brazil.Although numerous Salmonella outbreaks associated with melons have been reported in the United States and elsewhere, this is the first of its kind in Europe.Expansion of the melon import market from Brazil represents a potential threat for future outbreaks. Whole genome sequencing is rapidly becoming more accessible and can provide a compelling level of evidence of linkage between human cases and sources of infection,to support public health interventions in global food markets.


Diabetes-induced alteration in brain monoamine metabolism in rats.
Concentrations of monoamines and their metabolites as well as the activities of tyrosine hydroxylase (TH) and of choline acetyltransferase were investigated in various brain regions of control and streptozotocin-treated Sprague-Dawley rats. The animals were rendered diabetic by a single i.v. injection of streptozotocin (65 mg/kg) and killed 10, 30 and 90 days after the treatment. During the course of diabetes, progressive decreases in the activity of TH and a marked increase in the concentration of norepinephrine were observed in several brain regions, including thalamus, hypothalamus, medulla and midbrain. This inverse relationship between TH activity and norepinephrine content was also seen in the pons but only in the 90-day diabetic animals. Lower TH activity in the hypothalamus of 30-day diabetic rats reflected a decreased Vmax, but no difference in the Km. The number of alpha adrenergic receptors was increased significantly in the hypothalamus, medulla and midbrain of the 30-day diabetic rats. The concentrations of dopamine and serotonin in various brain regions of the 10- and 30-day diabetic rats were generally not significantly different from controls. Concentrations of the acidic metabolites of these neurotransmitters, dihydroxyphenylacetic acid and 5-hydroxyindolacetic acid, were, however, greatly reduced. The activity of choline acetyltransferase, a marker of presynaptic cholinergic neuron activity, remained unaltered during the course of diabetes. These data suggest that uncontrolled diabetes is associated with a significant disturbance of brain monoamine metabolism.


Rapid single-breath-hold 3D late gadolinium enhancement cardiac MRI using a stack-of-spirals acquisition.
To develop a rapid single-breath-hold 3D late gadolinium enhancement (LGE) magnetic resonance imaging (MRI) method, and demonstrate its feasibility in cardiac patients. An inversion recovery dual-density 3D stack-of-spirals imaging sequence was developed. The spiral acquisition was 2-fold accelerated by self-consistent parallel imaging reconstruction (SPIRiT), which resulted in a total scan time of 12 heartbeats. Field map-based linear off-resonance correction was incorporated to the SPIRiT reconstruction. The 3D spiral LGE scans were performed in 15 patients who were referred for clinically ordered cardiac MR examinations that included the standard 2D multislice LGE imaging. Image sharpness and overall quality were qualitatively assessed based on 5-point scales. Scar-induced hyper-LGE was identified in 4 out of the 15 patients by both 3D spiral and 2D multislice LGE tests. On average over all datasets (n = 15), the image sharpness scores were 3.9 (3D spiral) and 4.0 (2D multislice), and the image quality scores were 4.1 (3D spiral) and 4.0 (2D multislice) with no significant difference in both metrics (paired t-test; P > 0.1). The average scar contrast enhancement ratios were 0.72 and 0.75 in 3D and 2D images, respectively (n = 4). The average difference of fractional scar volumes measured in 3D and 2D images was 4.3% (n = 3). Stack-of-spiral acquisition combined with non-Cartesian SPIRiT parallel imaging enables rapid 3D LGE MRI in a 12 heartbeat-long breath-hold.J.


Improvement of UV electroluminescence of n-ZnO/p-GaN heterojunction LED by ZnS interlayer.
n-ZnO/p-GaN heterojunction light emitting diodes with different interfacial layers were fabricated by pulsed laser deposition. The electroluminescence (EL) spectra of the n-ZnO/p-GaN diodes display a broad blue-violet emission centered at 430 nm, whereas the n-ZnO/ZnS/p-GaN and n-ZnO/AlN/p-GaN devices exhibit ultraviolet (UV) emission. Compared with the AlN interlayer, which is blocking both electron and hole at hetero-interface, the utilization of ZnS as intermediate layer can lower the barrier height for holes and keep an effective blocking for electron. Thus, an improved UV EL intensity and a low turn-on voltage (~5V) were obtained. The results were studied by peak-deconvolution with Gaussian functions and were discussed using the band diagram of heterojunctions.


Polymorphisms of human N-acetyltransferases and cancer risk.
Human arylamine N-acetyltransferases (CoASAc; NAT, EC 22.214.171.124) NAT1 and NAT2 play a key role in the metabolism of drugs and environmental chemicals and in the metabolic activation and detoxification of procarcinogens. Phenotyping analyses have revealed an association between NAT enzyme activities and the risk of developing several forms of cancer. As genotyping procedures have become available for NAT1 and NAT2 gene variations, hundreds of association studies on NAT polymorphisms and cancer risk have been conducted. Here we review the findings obtained from these studies. Evidence for a putative association of NAT1 polymorphism and myeloma, lung and bladder cancer, as well as association of NAT2 polymorphisms with non-Hodgkin lymphoma, liver, colorectal and bladder cancer have been reported. In contrast, no consistent evidence for a relevant association of NAT polymorphisms with brain, head & neck, breast, gastric, pancreatic or prostate cancer have been described. Although preliminary data are available, further well-powered studies are required to fully elucidate the role of NAT1 in most human cancers, and that of NAT2 in astrocytoma, meningioma, esophageal, renal, cervical and testicular cancers, as well as in leukaemia and myeloma. This review discusses controversial findings on cancer risk and putative causes of heterogeneity in the proposed associations, and it identifies topics that require further investigation, particularly mechanisms underlying association of NAT polymorphisms and risk for subsets of cancer patients with specific exposures, putative epistatic contribution of polymorphism for other xenobiotic-metabolising enzymes such as glutathione S-transferases of Cytochrome P450 enzymes, and genetic plus environmental interaction.


Effects of monosodium-L-glutamate administration on serum levels of reproductive hormones and cholesterol, epididymal sperm reserves and testicular histomorphology of male albino rats.
This study investigated the effects of administration of monosodium L-glutamate (MSG) on serum gonadotrophin-releasing hormone (GnRH), luteinising hormone (LH), testosterone and total cholesterol (TC), cauda epididymal sperm reserves (CESR) and testicular histomorphology of adult male albino rats. Eighty-four rats, randomly assigned to 7 groups of 12 rats each, were used for the study. Varying low doses (0.25, 0.50 or 1.00 g/kg body weight) of MSG were administered orally or subcutaneously at 48-h intervals for six weeks. Serum GnRH, LH, testosterone and TC, and CESR were evaluated on days 14, 28 and 42 of MSG administration. Testicular histomorphology was evaluated on day 42. The results showed that the mean serum GnRH, LH and testosterone levels, and the CESR of all the treated groups were significantly (P < 0.05) lower than those of the untreated control on days 14, 28 and 42 of MSG administration. The mean serum TC levels of all the treated groups were also significantly (P < 0.05) lower than those of the control group on days 14 and 28. No lesions were observed on sections of the testes. It was concluded that MSG administration for 14, 28 and 42 days led to significantly lower serum levels of GnRH, LH, testosterone and TC, and significantly lower CESR.


Cell-Type-Specific Transcriptome Analysis in the Drosophila Mushroom Body Reveals Memory-Related Changes in Gene Expression.
Learning and memory formation in Drosophila rely on a network of neurons in the mushroom bodies (MBs). Whereas numerous studies have delineated roles for individual cell types within this network in aspects of learning or memory, whether or not these cells can also be distinguished by the genes they express remains unresolved. In addition, the changes in gene expression that accompany long-term memory formation within the MBs have not yet been studied by neuron type. Here, we address both issues by performing RNA sequencing on single cell types (harvested via patch pipets) within the MB. We discover that the expression of genes that encode cell surface receptors is sufficient to identify cell types and that a subset of these genes, required for sensory transduction in peripheral sensory neurons, is not only expressed within individual neurons of the MB in the central brain, but is also critical for memory formation.


Targeted Sequencing of Lung Function Loci in Chronic Obstructive Pulmonary Disease Cases and Controls.
Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide; smoking is the main risk factor for COPD, but genetic factors are also relevant contributors. Genome-wide association studies (GWAS) of the lung function measures used in the diagnosis of COPD have identified a number of loci, however association signals are often broad and collectively these loci only explain a small proportion of the heritability. In order to examine the association with COPD risk of genetic variants down to low allele frequencies, to aid fine-mapping of association signals and to explain more of the missing heritability, we undertook a targeted sequencing study in 300 COPD cases and 300 smoking controls for 26 loci previously reported to be associated with lung function. We used a pooled sequencing approach, with 12 pools of 25 individuals each, enabling high depth (30x) coverage per sample to be achieved. This pooled design maximised sample size and therefore power, but led to challenges during variant-calling since sequencing error rates and minor allele frequencies for rare variants can be very similar. For this reason we employed a rigorous quality control pipeline for variant detection which included the use of 3 independent calling algorithms. In order to avoid false positive associations we also developed tests to detect variants with potential batch effects and removed them before undertaking association testing. We tested for the effects of single variants and the combined effect of rare variants within a locus. We followed up the top signals with data available (only 67% of collapsing methods signals) in 4,249 COPD cases and 11,916 smoking controls from UK Biobank. We provide suggestive evidence for the combined effect of rare variants on COPD risk in TNXB and in sliding windows within MECOM and upstream of HHIP. These findings can lead to an improved understanding of the molecular pathways involved in the development of COPD.


Mycolactone: a polyketide toxin from Mycobacterium ulcerans required for virulence.
Mycobacterium ulcerans is the causative agent of Buruli ulcer, a severe human skin disease that occurs primarily in Africa and Australia. Infection with M. ulcerans results in persistent severe necrosis without an acute inflammatory response. The presence of histopathological changes distant from the site of infection suggested that pathogenesis might be toxin mediated. A polyketide-derived macrolide designated mycolactone was isolated that causes cytopathicity and cell cycle arrest in cultured L929 murine fibroblasts. Intradermal inoculation of purified toxin into guinea pigs produced a lesion similar to that of Buruli ulcer in humans. This toxin may represent one of a family of virulence factors associated with pathology in mycobacterial diseases such as leprosy and tuberculosis.


Cloning and expression of Aca f 1: a new allergen of Acacia farnesiana pollen.
Acacia farnesiana is the main source of allergenic pollen and one of the most important causes of respiratory allergic disease in tropical and subtropical regions of the world. The purpose of this study was to produce a recombinant variety of allergenic Ole e 1-like protein from the pollen of this tree. To predict its allergenic cross-reactivity with other members of the Ole e 1-like protein family of common allergenic plants, the nucleotide sequence homology of the Acacia Ole e 1-like protein was evaluated. Amplification of cDNA strands encoding Acacia Ole e 1-like protein was performed by polymerase chain reaction (PCR) and sequenced. Following expression in Escherichia coli using the pET-21b(+) vector, the recombinant protein was purified using metal-affinity chromatography. IgE-binding competence of purified recombinant Ole e 1- like protein (rAca f 1) was analysed by immunoassay using 25 sera collected from Acacia pollen-sensitised patients. Nucleotide sequencing revealed an open reading frame of 453 bp encoding 150 amino acid residues that belonged to the Ole e 1-like protein family, and 11 patients (44%) had considerable specific IgE levels for the rAca f 1. Immunodetection and inhibition assays indicated that the purified rAca f 1 may be the same as that in the crude extract. Aca f 1, the second allergen from Acacia pollen, was identified as a member of the family of Ole e 1-like protein. A high degree of homology was found among amino acid sequences of Aca f 1 and several allergenic members of Ole e 1-like protein family.


Imaging the square of the correlated two-electron wave function of a hydrogen molecule.
The toolbox for imaging molecules is well-equipped today. Some techniques visualize the geometrical structure, others the electron density or electron orbitals. Molecules are many-body systems for which the correlation between the constituents is decisive and the spatial and the momentum distribution of one electron depends on those of the other electrons and the nuclei. Such correlations have escaped direct observation by imaging techniques so far. Here, we implement an imaging scheme which visualizes correlations between electrons by coincident detection of the reaction fragments after high energy photofragmentation. With this technique, we examine the H2 two-electron wave function in which electron-electron correlation beyond the mean-field level is prominent. We visualize the dependence of the wave function on the internuclear distance. High energy photoelectrons are shown to be a powerful tool for molecular imaging. Our study paves the way for future time resolved correlation imaging at FELs and laser based X-ray sources.


Borrelia burgdorferi protein interactions critical for microbial persistence in mammals.
Borrelia burgdorferi is the causative agent of Lyme disease that persists in a complex enzootic life cycle, involving Ixodes ticks and vertebrate hosts. The microbe invades ticks and vertebrate hosts in spite of active immune surveillance and potent microbicidal responses, and establishes long-term infection utilising mechanisms that are yet to be unravelled. The pathogen can cause multi-system disorders when transmitted to susceptible mammalian hosts, including in humans. In the past decades, several studies identified a limited number of B. burgdorferi gene-products critical for pathogen persistence, transmission between the vectors and the host, and host-pathogen interactions. This review will focus on the interactions between B. burgdorferi proteins, as well as between microbial proteins and host components, protein and non-protein components, highlighting their roles in pathogen persistence in the mammalian host. A better understanding of the contributions of protein interactions in the microbial virulence and persistence of B. burgdorferi would support development of novel therapeutics against the infection.


Heterozygous nonsense SCN5A mutation W822X explains a simultaneous sudden infant death syndrome.
The sudden, unexpected, and unexplained death of both members of a set of healthy twins (simultaneous sudden infant death syndrome (SSIDS)) is defined as a case in which both infants meet the definition of sudden infant death syndrome individually. A search of the world medical literature resulted in only 42 reported cases of SSIDS. We report the case of a pair of identical, male, monozygotic twins, 138 days old, who suddenly died, meeting the full criteria of SSIDS and where a genetic screen was performed, resulting in a heterozygous nonsense SCN5A mutation (W822X) in both twins. Immunohistochemistry was performed on cardiac tissue samples utilizing polyclonal antibodies anti-Na+ CP type Valpha (C-20) and a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling assay. The cellular localization of the Na+ CP type Valpha (C-20) demonstrated by confocal microscopy on staining pattern of myocytes was concentrated in the intercalated disks of ventricular myocytes. These findings suggest that defective ion channels represent viable candidates for the pathogenesis of SIDS and, obviously, of SSIDS, supporting a link between sudden infant death syndrome and cardiac channelopathies.


Development of peripheral chemoreceptor function in infants with chronic lung disease and initially lacking hyperoxic response.
Ten preterm infants with chronic lung disease (CLD) and undeveloped peripheral chemoreceptor function, described as ventilatory response to hyperoxia, were investigated, according to an individual protocol. Each infant was followed up until the response to hyperoxic inhalation had been observed on two occasions. Each examination consisted of overnight recording of saturation, testing of lung compliance and airway resistance, and the hyperoxic test. The hyperoxic response appeared at a mean postnatal age of 14 weeks (range 9-33 weeks). This response, which was independent of the infant's lung mechanics, appeared much later in infants with the severe form of CLD. As undeveloped peripheral chemoreceptor function has been suggested to be a key factor in sudden infant death syndrome (SIDS), the delayed development of their chemosensitivity leaves some infants with CLD unprotected against hypoxia at the age at which the risk for SIDS is highest.


Dietary factors and the risk of incident kidney stones in men: new insights after 14 years of follow-up.
Diet plays an important role in the pathogenesis of kidney stones. Because the metabolism of many dietary factors, such as calcium, may change with age, the relation between diet and kidney stones may be different in older adults. Uncertainty also remains about the association between many dietary factors, such as vitamin C, magnesium, and animal protein, and the risk of kidney stone formation. To examine the association between dietary factors and the risk of incident, symptomatic kidney stones in men and to determine whether these associations vary with age, a prospective cohort study was conducted of 45,619 men without a history of nephrolithiasis. Self-administered food frequency questionnaires were used to assess diet every 4 yr. A total of 1473 incident symptomatic kidney stones were documented during 477,700 person-years of follow-up. For men aged <60 yr, the multivariate relative risk (RR) for stone formation in the highest quintile of dietary calcium as compared with the lowest quintile was 0.69 (95% confidence interval [CI], 0.56 to 0.87; P = 0.01 for trend). By contrast, there was no association between dietary calcium and stone formation in men aged 60 yr or older. The multivariate RR for men who consumed 1000 mg or greater of vitamin C per day compared with those who consumed less than the recommended dietary allowance of 90 mg/d was 1.41 (95% CI, 1.11 to 1.80; P = 0.01 for trend). Other dietary factors showed the following multivariate RR among men in the highest quintile of intake compared with those in the lowest: magnesium, 0.71 (95% CI, 0.56 to 0.89; P = 0.01 for trend); potassium, 0.54 (95% CI, 0.42 to 0.68; P < 0.001 for trend); and fluid, 0.71 (95% CI, 0.59 to 0.85; P < 0.001 for trend). Animal protein was associated with risk only in men with a body mass index <25 kg/m(2) (RR, 1.38; 95% CI, 1.05 to 1.81; P = 0.03 for trend). Sodium, phosphorus, sucrose, phytate, vitamin B(6), vitamin D, and supplemental calcium were not independently associated with risk. In conclusion, the association between calcium intake and kidney stone formation varies with age. Magnesium intake decreases and total vitamin C intake seems to increase the risk of symptomatic nephrolithiasis. Because age and body size affect the relation between diet and kidney stones, dietary recommendations for stone prevention should be tailored to the individual patient.


Little bluestem litter dynamics in Minnesota old fields.
We studied the decay and nitrogen dynamics of little bluestem (Schizachyrium scoparium) litter in fertilized and unfertilized Minnesota old fields, using the litterbag technique. Annual decay rates and nitrogen leaching losses during the first month of decay were highly correlated with N content of litter and not with fertilizer additions. After the first month of decay, nitrogen was immobilized for at least 18 months. In contrast to decay rates and early N leaching losses, these immobilization rates were correlated with the amount of ammonium nitrate added in fertilizer rather than with initial %N. Therefore, litter quality and exogenous nitrogen supply appear to have different and independent effects on decay rates and N dynamics of little bluestem litter.


Selective differential human blood bilayer media for isolation of Gardnerella (Haemophilus) vaginalis.
New selective and differential human blood bilayer agar media with Tween 80 (HBT medium) or without Tween 80 (HB medium), developed for the isolation of Gardnerella (Haemophilus) vaginalis, permitted significantly higher G. vaginalis isolation rates than have been obtained for other media used for this purpose. HB medium consists of a basal layer of Columbia agar base containing colistin and naladixic acid with added amphotericin B and an overlayer of the same composition plus 5% human blood. HBT agar also contains Proteose Peptone No. 3 (Difco Laboratories) and Tween 80 in the basal layer and the overlayer. Both Tween 80 and the bilayer composition enhanced G. vaginalis production of human blood hemolysis, permitting detection of this organism even in the presence of heavy growth of other vaginal flora. The use of HB or HBT medium thus permitted the demonstration that G. vaginalis was present in vaginal fluid from a large percentage (up to 68%) of normal women. However, the concentration of G. vaginalis was found by semiquantitative analysis to be significantly higher in vaginal fluid from women with nonspecific vaginitis than in fluid from normal women.


Compositional epistasis: an epidemiologic perspective.
Under Bateson's original conception, the term "epistasis" is used to describe the situation in which the effect of a genetic factor at one locus is masked by a variant at another locus. Epistasis in the sense of masking has been termed "compositional epistasis." In general, statistical tests for interaction are of limited use in detecting compositional epistasis. Using recently developed epidemiological methods, however, it has been shown that there are relations between empirical data patterns and compositional epistasis. These relations can sometimes be exploited to empirically test for certain forms of compositional epistasis, by using alternative nonstandard tests for interaction.Using the counterfactual framework, we show conditions that can be empirically tested to determine whether there are individuals whose phenotype response patterns manifest epistasis in the sense of masking. Only under some very strong assumptions would tests for standard statistical interactions correspond to compositional epistasis. Even without such strong assumptions, however, one can still test whether there are individuals of phenotype response type representing compositional epistasis. The empirical conditions are quite strong, but the conclusions which tests of these conditions allow may be of interest in a wide range of studies. This chapter highlights that epidemiologic perspectives can be used to shed light on underlying mechanisms at the genetic, molecular, and cellular levels.


A biopsychosocial model of interferon-alpha-induced depression in patients with chronic hepatitis C infection.
The aim of this prospective study was to gain a more comprehensive picture of the biopsychosocial effects of interferon-α (IFN-α) treatment of patients with chronic hepatitis C (HCV). The predictors of depressive development and changes in health-related quality of life, life satisfaction and cognitive ability were measured with the inclusion of the social context. Furthermore, the effects of IFN-α treatment on indoleamine 2,3-dioxygenase, the level of tryptophan supply in the brain, the development of neurotoxic kynurenine metabolites and the thyroid glands were investigated. Therefore, for the first time the conditions for the development of depressive episodes in HCV patients treated with IFN-α were examined over the entire period of treatment as well as 3 months later, applying a holistic biopsychosocial model. Psychiatric and biological assessments were carried out at 6 different times: before, during (at 1, 3, 6 and 9 months) and after the end of IFN-α treatment. During IFN-α treatment 22 (53.7%) of 41 patients fulfilled the criteria for a treatment-related depressive disorder at least once during treatment. Contributing factors are tryptophan depletion (tryptophan to competing amino acids quotient), increased neurotoxic challenge (kynurenine to kynurenic acid quotient), less social support, female gender, preexisting psychiatric vulnerability, means of transmission, low financial security, impaired sexual satisfaction, small circle of friends, impaired physical role, strong body pain, low general health and vitality, reduced social functioning, impaired mental health and impaired emotional role. The awareness of relevant risk factors of IFN-α treatment-induced depression is essential to develop preventative treatment strategies.


Whole genome sequencing for investigations of meningococcal outbreaks in the United States: a retrospective analysis.
Although rare in the U.S., outbreaks due to Neisseria meningitidis do occur. Rapid, early outbreak detection is important for timely public health response. In this study, we characterized U.S. meningococcal isolates (N = 201) from 15 epidemiologically defined outbreaks (2009-2015) along with temporally and geographically matched sporadic isolates using multilocus sequence typing, pulsed-field gel electrophoresis (PFGE), and six whole genome sequencing (WGS) based methods. Recombination-corrected maximum likelihood (ML) and Bayesian phylogenies were reconstructed to identify genetically related outbreak isolates. All WGS analysis methods showed high degree of agreement and distinguished isolates with similar or indistinguishable PFGE patterns, or the same strain genotype. Ten outbreaks were caused by a single strain; 5 were due to multiple strains. Five sporadic isolates were phylogenetically related to 2 outbreaks. Analysis of 9 outbreaks using timed phylogenies identified the possible origin and estimated the approximate time that the most recent common ancestor emerged for outbreaks analyzed. U.S. meningococcal outbreaks were caused by single- or multiple-strain introduction, with organizational outbreaks mainly caused by a clonal strain and community outbreaks by divergent strains. WGS can infer linkage of meningococcal cases when epidemiological links are uncertain. Accurate identification of outbreak-associated cases requires both WGS typing and epidemiological data.


Reconstitution of Ca2+-regulated membrane fusion by synaptotagmin and SNAREs.
We investigated the effect of synaptotagmin I on membrane fusion mediated by neuronal SNARE proteins, SNAP-25, syntaxin, and synaptobrevin, which were reconstituted into vesicles. In the presence of Ca2+, the cytoplasmic domain of synaptotagmin I (syt) strongly stimulated membrane fusion when synaptobrevin densities were similar to those found in native synaptic vesicles. The Ca2+ dependence of syt-stimulated fusion was modulated by changes in lipid composition of the vesicles and by a truncation that mimics cleavage of SNAP-25 by botulinum neurotoxin A. Stimulation of fusion was abolished by disrupting the Ca2+-binding activity, or by severing the tandem C2 domains, of syt. Thus, syt and SNAREs are likely to represent the minimal protein complement for Ca2+-triggered exocytosis.


Elevated levels of urinary prostaglandin e metabolite indicate a poor prognosis in ever smoker head and neck squamous cell carcinoma patients.
Cyclooxygenase (COX)-derived prostaglandin E(2) (PGE(2)) plays a role in the development and progression of several tumor types including head and neck squamous cell carcinoma (HNSCC). Measurements of urinary PGE metabolite (PGE-M) can be used as an index of systemic PGE(2) production. In ever smokers, increased levels of urinary PGE-M reflect increased COX-2 activity. In this study, we determined whether baseline levels of urinary PGE-M were prognostic for ever smoker HNSCC patients. A retrospective chart review of ever smoker HNSCC patients treated with curative intent was done. Fifteen of 31 evaluable patients developed progressive disease (recurrence or a second primary tumor) after a median follow-up of 38 months. There were no statistically significant differences between patients with (n = 15) or without disease progression (n = 16) with regard to stage, site, treatment received, smoking status, and aspirin use during follow-up. Median urinary PGE-M levels were significantly higher in HNSCC patients with disease progression (21.7 ng/mg creatinine) compared with patients without (13.35 ng/mg creatinine; P = 0.03). Importantly, patients with high baseline levels of urinary PGE-M had a significantly greater risk of disease progression (hazard ratio, 4.76, 95% CI, 1.31-17.30; P < 0.01) and death (hazard ratio, 9.54; 95% CI, 1.17-77.7; P = 0.01) than patients with low baseline levels of urinary PGE-M. These differences were most evident among patients with early-stage disease. Taken together, our findings suggest that high baseline levels of urinary PGE-M indicate a poor prognosis in HNSCC patients. Possibly, HNSCC patients with high COX-2 activity manifested by elevated urinary PGE-M will benefit from treatment with a COX-2 inhibitor.


Causes of Alzheimer's disease.
It is now understood that genetic factors play a crucial role in the risk of developing Alzheimer's disease (AD). Rare mutations in at least 3 genes are responsible for early-onset familial AD. A common polymorphism in the apolipoprotein E gene is the major determinant of risk in families with late-onset AD, as well as in the general population. Advanced age, however, remains the major established risk factor for AD, although environmental variables may also have some role in disease expression. Some pathogenic factors directly associated with aging include oxidative damage and mutations in messenger RNA. Other factors unrelated to the aging process may, in the future, be amenable to therapeutic intervention by way of estrogen replacement therapy for postmenopausal women, anti-inflammatory drug therapy and reducing vascular risk factors. Older theories, such as aluminum playing a role in the pathogenesis of AD, have been mostly discarded as our understanding of pathogenic mechanisms of AD has advanced.


Helping consumers achieve recommended intakes of whole grain foods.
According to research data from both the USDA and private companies, Americans consume approximately one serving of whole grain foods per day. Current guidelines recommend three or more servings a day. The health benefits of whole grains are well documented. However, convincing Americans to eat them, especially in the amount recommended, has been unsuccessful. Adults and children give a variety of reasons for not choosing whole grain foods, including these: a lack of understanding of the health benefits of whole grain foods, an inability to identify whole grain foods at the point of purchase, their taste, their cost, the time it takes to prepare them and the availability of whole grain foods in stores. This paper addresses some of the misconceptions surrounding whole grain foods and offers strategies for helping consumers choose more whole grain products. In addition, we present historical information on the consumption trends of whole grains as well as educational and promotional opportunities that industry and health professionals can use to encourage the public to eat more whole grains.


On rare-variant analysis in population-based designs: decomposing the likelihood to two informative components.
Various analytical approaches have been suggested for the characterization of rare variants. One main approach is to collapse the genetic information of rare variants in a region and to construct an overall test statistic. Here, we proposed a new approach based on collapsed genotype scores. By utilizing the information of the association signal that is ignored in collapsing methods, i.e. the configuration of rare alleles, we constructed a more powerful test and compared it with existing rare-variant approaches. With extensive simulation studies, we showed that our method performs better than existing approaches, and we applied our method to a sequencing study of nonsyndromic cleft lip illustrating the practical advantages of the proposed method.


Comparison of the performance of normal individuals and survivors of traumatic brain injury on repeat administrations of the Wisconsin Card Sorting Test.
Repeat administrations of the Wisconsin Card Sorting Test (WCST) are a common clinical practice to assess changes in cognitive functions of individuals recovering from traumatic brain injury (TBI). Yet, little is even known about the performance of healthy individuals on repeat administrations of the test. The present study compared the effects of repeat administration of the WCST in normal individuals and survivors of traumatic brain injury. Normal participants were administered the WCST twice, with a 5-month interval between testings and the findings were compared to the data from patients with TBI tested twice. Normal controls only modestly improved their performance on repeat testing. In contrast, using a reliability of change (RC) index, TBI survivors demonstrated clinically meaningful gains in performance.


An update on primary sclerosing cholangitis.
Primary sclerosing cholangitis is a chronic cholestatic liver disease characterized by strictures of the biliary tree complicated by cirrhosis and cholangiocarcinoma. It is immune mediated, although the precise aetiology remains unknown. The research into aetiology, genetic associations, pathogenesis, epidemiology, diagnosis of cholangiocarcinoma and medical treatments are discussed. Multiple gene polymorphisms and human leucocyte antigen haplotype associations with primary sclerosing cholangitis have been investigated. Common inflammatory bowel disease associated polymorphisms and ulcerative colitis associated human leucocyte antigen haplotypes are not associated with primary sclerosing cholangitis. Biliary epithelial cells may mediate their own destruction by exaggerating innate and adaptive immune responses to bacterial products in the liver. The natural history of large and small duct primary sclerosing cholangitis has been reviewed. Positron emission tomography may be a useful adjunct to current imaging modalities in the pretransplant assessment of patients to exclude cholangiocarcinoma. Ursodeoxycholic acid remains the most studied medical treatment for primary sclerosing cholangitis; pilot studies suggest a possible role for tacrolimus and silymarin, however further studies are required.


Effect of avocado/soybean unsaponifiables on ligature-induced bone loss and bone repair after ligature removal in rats.
The aim of this study was to evaluate the effects of administration of avocado/soybean unsaponifiable (ASU), a drug that is commonly used in the treatment of rheumatoid arthritis, on ligature-induced bone loss and bone repair after ligature removal in rats. Eighty-four rats were randomly assigned to four groups of equal size and received a daily gavage of either sterile saline [control (CTR)] or ASU (0.6 mg/kg), starting 7 d before (ASU/-7), on the day of (ASU/0) or 7 d after (ASU/+7) periodontitis induction. Periodontitis was induced by placing silk ligatures into the gingival sulcus of the second maxillary molars for 7 d; after 7 d, the ligatures were removed. Seven rats from each group were sacrificed, 7, 15 or 30 d after ligature removal. Bone resorption was evaluated by histomorphometry and micro-computed tomography (micro-CT). Immunohistochemistry was used to evaluate the expression of TRAP, RANKL and alkaline phosphatase (ALP), and quantitative PCR (qPCR) was used to evaluate the levels of interleukin-1beta (Il1β), tumor necrosis factor alpha (Tnfα), interleukin-6 (Il-6), Rankl and Alp. Statistical analysis was performed using the Shapiro-Wilk test, ANOVA and Tukey's test for normal data, and using the Kruskall-Wallis and Dunnet's tests for non-normal data (p < 0.05). Histomorphometry and micro-CT analysis showed greater bone resorption in the CTR group than in the ASU/0 (15 d) and ASU/+7 (7 and 15 d) groups. The CTR group also presented with a higher expression of TRAP (15 and 30 d) and RANKL (7 and 15 d) compared with ASU/0 and ASU/+7 groups. Similarly, qPCR analysis showed higher levels of Rankl and Il1β mRNAs, and lower levels of Alp mRNA, in the CTR group compared with all other groups (for all periods). ASU exhibited a positive effect on bone repair following ligature-induced periodontitis in rats.


Long-term survival and function of intrahepatic islet allografts in baboons treated with humanized anti-CD154.
Clinical islet cell transplantation has resulted in insulin independence in a limited number of cases. Rejection, recurrence of autoimmunity, and impairment of normal islet function by conventional immunosuppressive drugs, e.g., steroids, tacrolimus, and cyclosporin A, may all contribute to islet allograft loss. Furthermore, intraportal infusion of allogeneic islets results in the activation of intrahepatic macrophages and endothelial cells, followed by production of proinflammatory mediators that can contribute to islet primary nonfunction. We reasoned that the beneficial effects of anti-CD154 treatment on autoimmunity, alloreactivity, and proinflammatory events mediated by macrophages and endothelial cells made it an ideal agent for the prevention of islet allograft failure. In this study, a nonhuman primate model (Papio hamadryas) was used to assess the effect of humanized anti-CD154 (hu5c8) on allogeneic islet engraftment and function. Nonimmunosuppressed and tacrolimus-treated recipients were insulin independent posttransplant, but rejected their islet allografts in 8 days. Engraftment and insulin independence were achieved in seven of seven baboon recipients of anti-CD154 induction therapy administered on days -1, 3, and 10 relative to the islet transplant. Three of three baboons treated with 20 mg/kg anti-CD154 induction therapy experienced delayed rejection episodes, first detected by elevations in postprandial blood glucose levels, on postoperative day (POD) 31 for one and on POD 58 for the other two. Re-treatment with three doses of anti-CD154 resulted in reversal of rejection in all three animals and in a return to normoglycemia and insulin independence in two of three baboons. It was possible to reverse multiple episodes of rejection with this approach. A loss of functional islet mass, as detected by reduced first-phase insulin release in response to intravenous glucose tolerance testing, was observed after each episode of rejection. One of two baboons treated with 10 mg/kg induction therapy became insulin independent post-transplant but rejected the islet graft on POD 10; the other animal experienced a reversible rejection episode on POD 58 and remained insulin independent and normoglycemic until POD 264. Two additional baboon recipients of allogeneic islets and donor bone marrow (infused on PODs 5 and 11) were treated with induction therapy (PODs -1, 3, 10), followed by initiation of monthly maintenance therapy (for a period of 6 months) on POD 28. Rejection-free graft survival and insulin independence was maintained for 114 and 238 days, with preservation of functional islet mass observed in the absence of rejection. Prevention and reversal of rejection, in the absence of the deleterious effects associated with the use of conventional immunosuppressive drugs, make anti-CD154 a unique agent for further study in islet cell transplantation.


Inhibition of Na+/H+ exchange stimulates CCK secretion in STC-1 cells.
It has been demonstrated that K+ channel regulation of membrane potential is critical for control of CCK secretion. Because certain K+ channels are pH sensitive, it was postulated that pH affects K+ channel activity in the CCK-secreting cell line STC-1 and may participate in regulating CCK secretion. The present study examines the role of electroneutral Na+/H+ exchange on extracellular acidification and hormone secretion. Treatment of STC-1 cells with the amiloride analog ethylisopropyl amiloride (EIPA) to inhibit Na+/H+ exchange inhibited Na+-dependent H+ efflux and increased basal CCK secretion. Substituting choline for NaCl in the extracellular medium elevated basal intracellular Ca2+ concentration and stimulated CCK release. Stimulatory effects on hormone secretion were blocked by the L-type Ca2+ channel blocker diltiazem, indicating that secretion was dependent on the influx of extracellular Ca2+. To determine whether the effects of EIPA and Na+ depletion were due to membrane depolarization, we tested graded KCl concentrations. The ability of EIPA to increase CCK secretion was inhibited by depolarization induced by 10-50 mM KCl in the bath. Maneuvers to lower intracellular pH (pHi), including reducing extracellular pH (pHo) to 7.0 or treatment with sodium butyrate, significantly increased CCK secretion. To examine whether pH directly affects membrane K+ permeability, we measured outward currents carried by K+, using whole cell patch techniques. K+ current was significantly inhibited by lowering pHo to 7.0. These effects appear to be mediated through changes in pHi, because intracellular dialysis with acidic solutions nearly eliminated current activity. These results suggest that Na+/H+ exchange and membrane potential may be functionally linked, where inhibition of Na+/H+ exchange lowers pHi and depolarizes the membrane, perhaps through inhibition of pH-sensitive K+ channels. In turn, K+ channel closure and membrane depolarization open voltage-dependent Ca2+ channels, leading to an increase in cytosolic Ca2+ and CCK release. The effects of pHi on K+ channels may serve as a potent stimulus for hormone secretion, linking cell metabolism and secretory functions.


Generic model abstraction from examples.
The recognition community has typically avoided bridging the representational gap between traditional, low-level image features and generic models. Instead, the gap has been artificially eliminated by either bringing the image closer to the models using simple scenes containing idealized, textureless objects or by bringing the models closer to the images using 3D CAD model templates or 2D appearance model templates. In this paper, we attempt to bridge the representational gap for the domain of model acquisition. Specifically, we address the problem of automatically acquiring a generic 2D view-based class model from a set of images, each containing an exemplar object belonging to that class. We introduce a novel graph-theoretical formulation of the problem in which we search for the lowest common abstraction among a set of lattices, each representing the space of all possible region groupings in a region adjacency graph representation of an input image. The problem is intractable and we present a shortest path-based approximation algorithm to yield an efficient solution. We demonstrate the approach on real imagery.


Impact of an intensive lifestyle intervention on use and cost of medical services among overweight and obese adults with type 2 diabetes: the action for health in diabetes.
To assess the relative impact of an intensive lifestyle intervention (ILI) on use and costs of health care within the Look AHEAD trial. A total of 5,121 overweight or obese adults with type 2 diabetes were randomly assigned to an ILI that promoted weight loss or to a comparison condition of diabetes support and education (DSE). Use and costs of health-care services were recorded across an average of 10 years. ILI led to reductions in annual hospitalizations (11%, P = 0.004), hospital days (15%, P = 0.01), and number of medications (6%, P < 0.001), resulting in cost savings for hospitalization (10%, P = 0.04) and medication (7%, P < 0.001). ILI produced a mean relative per-person 10-year cost savings of $5,280 (95% CI 3,385-7,175); however, these were not evident among individuals with a history of cardiovascular disease. Compared with DSE over 10 years, ILI participants had fewer hospitalizations, fewer medications, and lower health-care costs.


Cell-free circulating plasma hTERT mRNA is a useful marker for prostate cancer diagnosis and is associated with poor prognosis tumor characteristics.
Serum prostate-specific antigen (PSA) is the most widely used marker for diagnosing prostate cancer (PCa). It lacks specificity and predictive value, resulting in inaccurate diagnoses and overtreatment of the disease. The aim of this study was to assess the usefulness of plasma telomerase reverse transcriptase (hTERT) mRNA as a diagnostic and prognostic tool for PCa and its association with clinicopathological parameters of tumors. Plasma hTERT mRNA levels were determined by qRT-PCR in 105 consecutive patients with elevated PSA levels and in 68 healthy volunteers. The diagnostic accuracy, the efficacy as a prognostic factor of biochemical recurrence and the association with tumor clinicopathological parameters of plasma hTERT mRNA and serum PSA tests were determined using univariate and multivariate analyses. The results show that plasma hTERT mRNA is a non-invasive biomarker for PCa diagnosis that shows higher sensitivity (85% vs. 83%), specificity (90% vs. 47%), positive predictive value (83% vs. 56%), and negative predictive value (92% vs. 77%) than serum PSA. Plasma hTERT mRNA is significantly associated with poor prognosis tumor clinicopathological parameters and is a significant independent predictor of PCa (p<0.0001). Univariate analysis identified plasma hTERT mRNA (but not serum PSA) as a significant prognostic factor of biochemical recurrence. Plasma hTERT mRNA Kaplan-Meier curves confirmed the significant differences between groups and patients with higher levels than the cut-off value showed diminished recurrence-free survival (p=0.004), whereas no differences were observed with serum PSA (p=0.38). Multivariate analysis indicated that plasma hTERT mRNA (but not serum PSA) and stage were significantly associated with biochemical recurrence. Overall, these findings indicate that hTERT mRNA is a useful non-invasive tumor marker for the molecular diagnosis of PCa, affording a greater diagnostic and prognostic accuracy than the PSA assay and may be of relevance in the follow-up of the disease.


An optimized kit-free method for making strand-specific deep sequencing libraries from RNA fragments.
Deep sequencing of strand-specific cDNA libraries is now a ubiquitous tool for identifying and quantifying RNAs in diverse sample types. The accuracy of conclusions drawn from these analyses depends on precise and quantitative conversion of the RNA sample into a DNA library suitable for sequencing. Here, we describe an optimized method of preparing strand-specific RNA deep sequencing libraries from small RNAs and variably sized RNA fragments obtained from ribonucleoprotein particle footprinting experiments or fragmentation of long RNAs. Our approach works across a wide range of input amounts (400 pg to 200 ng), is easy to follow and produces a library in 2-3 days at relatively low reagent cost, all while giving the user complete control over every step. Because all enzymatic reactions were optimized and driven to apparent completion, sequence diversity and species abundance in the input sample are well preserved.


Prevalence of hypovitaminosis D and the different-dose cholecalciferol supplementation effects on renal transplant recipients.
High prevalence of hypovitaminosis D has been observed in patients with chronic kidney disease. However, there is not much data about its prevalence in kidney transplant recipients (KTRs). The study included 83 adult KTRs at a single center to calculate the prevalence of hypovitaminosis D. Among the 83 patients, those with incomplete data were excluded leaving 22 patients available for study. The demographic and biochemical data were analyzed retrospectively. Serum concentrations of 25-hydroxyvitamin D [25(OH)D], parathyroid hormone (PTH), phosphorus, calcium, and creatinine were evaluated. The 22 selected patients were divided into two groups: (1) those who received 10,000 IU of cholecalciferol orally per week, and (2) those who received 10,000-20,000 IU/week. The Vitamin D level rate was calculated to evaluate the time necessary to reach serum values ≥30 ng/mL. Hypovitaminosis D was present in 80.7% (67/83) of the patients. Eleven patients received 10,000 IU/week of cholecalciferol, and the other 11 patients received 10,000-20,000 IU/week (approximately 64,000 IU/month). The calcium, phosphorus, and PTH values did not show any differences between the two groups. We estimate that a dose of approximately 64,000 IU/month of cholecalciferol was sufficient to reach values of ≥30 ng/mL of 25(OH)D in approximately 2.1 months in the insufficient and 4.3 months in Vitamin D-deficient patients. The prevalence of hypovitaminosis D was high among Brazilian KTR, and low-level doses of cholecalciferol (approximately 64,000 IU/month) were sufficient to control hypovitaminosis D.


Genome-wide association study of obsessive-compulsive disorder.
Obsessive-compulsive disorder (OCD) is a common, debilitating neuropsychiatric illness with complex genetic etiology. The International OCD Foundation Genetics Collaborative (IOCDF-GC) is a multi-national collaboration established to discover the genetic variation predisposing to OCD. A set of individuals affected with DSM-IV OCD, a subset of their parents, and unselected controls, were genotyped with several different Illumina SNP microarrays. After extensive data cleaning, 1465 cases, 5557 ancestry-matched controls and 400 complete trios remained, with a common set of 469,410 autosomal and 9657 X-chromosome single nucleotide polymorphisms (SNPs). Ancestry-stratified case-control association analyses were conducted for three genetically-defined subpopulations and combined in two meta-analyses, with and without the trio-based analysis. In the case-control analysis, the lowest two P-values were located within DLGAP1 (P=2.49 × 10(-6) and P=3.44 × 10(-6)), a member of the neuronal postsynaptic density complex. In the trio analysis, rs6131295, near BTBD3, exceeded the genome-wide significance threshold with a P-value=3.84 × 10(-8). However, when trios were meta-analyzed with the case-control samples, the P-value for this variant was 3.62 × 10(-5), losing genome-wide significance. Although no SNPs were identified to be associated with OCD at a genome-wide significant level in the combined trio-case-control sample, a significant enrichment of methylation QTLs (P<0.001) and frontal lobe expression quantitative trait loci (eQTLs) (P=0.001) was observed within the top-ranked SNPs (P<0.01) from the trio-case-control analysis, suggesting these top signals may have a broad role in gene expression in the brain, and possibly in the etiology of OCD.


High-content analysis of tumour cell invasion in three-dimensional spheroid assays.
Targeting infiltrating tumour cells is an attractive way of combating cancer invasion and metastasis. Here we describe a novel and reproducible method for high content analysis of invading cells using multicellular tumour spheroid assays in a high grade glioma model. Live cell imaging of spheroids generated from glioma cell lines, U87 and U251, gave insight into migration dynamics and cell morphology in response to anti-migratory drugs. Immunofluorescence imaging confirmed cytoskeletal rearrangements in the treated cells indicating a direct effect on cell morphology. Effect on migration was determined by a Migration Index (MI) from brightfield images which confirmed anti-migratory activity of the drugs. A marked effect on the core with treatment suggestive of disordered proliferation was also observed. A newly developed technique to prepare the spheroids and migratory cells for immunohistochemistry allowed an assessment of response to drug treatment with a selection of markers. A difference in protein expression was noted between cells maintained within the core and migratory cells indicative of the presence of cell subpopulations within the spheroid core. We conclude that this high content analysis allows researchers to perform screening of anti-tumour invasion compounds and study their effects on cellular dynamics, particularly in relation to protein expression, for the first time.


Prevalence of CYP1B1 mutations in Australian patients with primary congenital glaucoma.
Analysis of CYP1B1 in primary congenital glaucoma (PCG) patients from various ethnic populations indicates that allelic heterogeneity is high, and some mutations are population specific. No study has previously reported the rate or spectrum of CYP1B1 mutations in Australian PCG patients. The aim of this study is to determine the frequency of CYP1B1 mutations in our predominately Caucasian, Australian cohort of PCG cases. Thirty-seven probands were recruited from South-Eastern Australia, along with 100 normal control subjects. Genomic DNA was extracted and the coding regions of CYP1B1 analysed by direct sequencing. Sequence analysis identified 10 different CYP1B1 disease-causing variants in eight probands (21.6%). Five subjects were compound heterozygotes, two subjects heterozygous and one homozygous for CYP1B1 mutations. Three missense mutations are novel (D192Y, G329D, and P400S). None of the novel mutations identified were found in normal controls. One normal control subject was heterozygous for the previously reported CYP1B1 R368H mutation. Six previously described probable polymorphisms were also identified. Mutations in CYP1B1 account for approximately one in five PCG cases from Australia. Our data also supported the high degree of allelic heterogeneity seen in similar studies from other ethnic populations, thereby underscoring the fact that other PCG-related genes remain to be identified.


Effects of triclosan on the cytotoxicity and fungal growth on a soft denture liner.
Contamination of removable prostheses with microorganisms, particularly Candida albicans, is a common clinical problem. Microban, a broad-spectrum antimicrobial containing triclosan, recently has been proposed to inhibit microbial growth. This study aimed to determine whether the addition of Microban to PermaSoft denture liner prevents the growth of C albicans and affects the cytotoxicity of the PermaSoft material. Experimental specimen disks (5 x 1 mm each) with and without incorporated Microban were fabricated aseptically (n = 6) against polyester film to produce a smooth surface. To assess the cytotoxic effect of Microban, the MTT assay was used. To determine the effect of Microban on the growth of C albicans, disks were placed in Transwell dishes, covered with Sabouraud's broth containing an ATCC strain of C albicans, and incubated at 37 degrees C for 24 hours. Wells containing fluorocarbon resin disks or broth alone served as controls. The disks were rinsed to remove unattached C albicans and then sonicated in sterile water to remove surface organisms. Serial dilutions of the water extracts were plated on Sabouraud's agar and returned to the incubator for 24 hours. Colonies were counted with a Brunswick Colony Counter. Growth of C albicans in the internal aspects of the specimens was determined in a manner as previously described, with the exception that the specimens were sonicated to remove surface organisms, minced, and sonicated once more before making serial dilutions. The results were compared with ANOVA and Tukey intervals (alpha=.05). The number of colonies formed ranged from 17 to 31 x 10(5) (mean = 23 +/- 4 x 10(5)) and 14 to 69 x 10(5) (mean = 32 +/- 20 x 10(5)) for the PermaSoft with and without Microban groups, respectively. There was no statistically significant difference between PermaSoft with and without Microban. The addition of Microban did not significantly alter the cytotoxicity of the PermaSoft denture lining material or reduce the adherence of viable C albicans to the surface of PermaSoft material after 24 hours.


Effects of pink bollworm resistance to Bacillus thuringiensis on phenoloxidase activity and susceptibility to entomopathogenic nematodes.
Widespread planting of crops genetically engineered to produce insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) imposes selection on many key agricultural pests to evolve resistance to Bt. Fitness costs can slow the evolution of Bt resistance. We examined effects of entomopathogenic nematodes on fitness costs of Bt resistance in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), a major pest of cotton, Gossypium hirsutum L., in the southwestern United States that is currently controlled by transgenic cotton that produces Bt toxin Cry1Ac. We tested whether the entomopathogenic nematodes Steinernema riobrave Cabanillas, Poinar, and Raulston (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) affected fitness costs of resistance to Cry1Ac in two laboratory-selected hybrid strains of pink bollworm reared on non-Bt cotton bolls. The nematode S. riobrave imposed a recessive fitness cost for one strain, and H. bacteriophora imposed a fitness cost affecting heterozygous resistant individuals for the other strain. Activity of phenoloxidase, an important component of insects' immune response, did not differ between Bt-resistant and Bt-susceptible families. This suggests phenoloxidase does not affect susceptibility to entomopathogenic nematodes in Bt-resistant pink bollworm. Additionally, phenoloxidase activity does not contribute to Bt resistance, as has been found in some species. We conclude that other mechanisms cause higher nematode-imposed mortality for pink bollworm with Bt resistance genes. Incorporation of nematode-imposed fitness costs into a spatially explicit simulation model suggests that entomopathogenic nematodes in non-Bt refuges could delay resistance by pink bollworm to Bt cotton.


The necessity of continuous international cooperation for establishing the coronavirus disease 2019 diagnostic capacity despite the challenges of fighting the outbreak in home countries.
The importance of laboratory diagnostic capacity for effective infectious disease control has been widely recognized in recent years, but many of the countries still struggled to establish it when the newly discovered diseases was happened, such as coronavirus disease 2019 (COVID-19). Even in the country that the laboratory system was highly evaluated by Global Health Security Index like Myanmar, support from external partners is essential to establish the diagnostic capacity for COVID-19. WHO and other contributors, including Japan, have been supporting the establishment of a diagnostic system for SARS-CoV-2 in response to the disease outbreak. The testing laboratory was established in Myanmar on February 20, 2020. The first confirmed diagnosis was reported on March 23, and 15 positive cases as of March 31. Since it is difficult to control the outbreak in a given country without controlling it in the neighboring countries, continuous international cooperation for establishing the coronavirus disease 2019 diagnostic capacity was crucial despite the challenges of fighting the outbreak in home countries.


Challenges, Opportunities, and Methods for Large-Scale Evaluations.
The logic, theory, and practice of large-scale evaluation were once limited to large federal initiatives. However, with the advent of regularly collected performance measures and the often multisite implementation of quality improvement efforts, there is an opportunity to adopt large-scale evaluation methods in local and regional evaluation efforts. While ineffective programs show little variation in their ineffectiveness, effective programs generally show a range of effects. A central task of large-scale evaluation is to describe and ascribe why the same program, implemented in multiple settings, produces different effects. By its attention to variation attributable to setting, activities, outputs, and by participants and documenting the conditions in which programs achieve greater and lesser success, large-scale evaluation supports the needs of decision-makers when choosing to implement an evidence-based program. In addition to knowing a program is effective, decision-makers want to know whether it is appropriate for their situation and what facilitates or impedes effective implementation and bears on the program's ultimate effectiveness. This article presents the different methods and approaches appropriate for effectively and efficiently constructing and executing a large-scale evaluation that will provide decision-makers the evidence they need for evidence-informed adoption of effective programs.


The validity and scalability of the Theory of Mind Scale with toddlers and preschoolers.
Despite the importance of theory of mind (ToM) for typical development, there remain 2 key issues affecting our ability to draw robust conclusions. One is the continued focus on false belief as the sole measure of ToM. The second is the lack of empirically validated measures of ToM as a broad construct. Our key aim was to examine the validity and reliability of the 5-item ToM scale (Peterson, Wellman, & Liu, 2005). In particular, we extended on previous research of this scale by assessing its scalability and validity for use with children from 2 years of age. Sixty-eight typically developing children (aged 24 to 61 months) were assessed on the scale's 5 tasks, along with a sixth Sally-Anne false-belief task. Our data replicated the scalability of the 5 tasks for a Rasch-but not Guttman-scale. Guttman analysis showed that a 4-item scale may be more suitable for this age range. Further, the tasks showed good internal consistency and validity for use with children as young as 2 years of age. Overall, the measure provides a valid and reliable tool for the assessment of ToM, and in particular, the longitudinal assessment of this ability as a construct.


Management of hydrocephalus in the patient with myelomeningocele: an argument against third ventriculostomy.
The majority of children with myelomeningocele will have associated hydrocephalus. The management of hydrocephalus can be one of the most trying problems in this patient population. Cerebrospinal fluid (CSF) diversion will be required in these children for the remainder of their lives. Blockage of the outlets of the fourth ventricle and communication of the fourth ventricle with the central canal provides a mechanism for compensation. The signs and symptoms of CSF diversion malfunction, either shunt or third ventriculostomy, can be quite subtle. The objective indications of these malfunctions are less available after third ventriculostomy than when using mechanical shunting. The ease with which the diagnosis of malfunction can be made becomes the major advantage of mechanical shunting over third ventriculostomy.


Adjunctive remifentanil infusion in deeply sedated and paralyzed ICU patients during fiberoptic bronchoscopy procedure: a prospective, randomized, controlled study.
Even with an adequate pain assessment, critically ill patients under sedation experience pain during procedures in the intensive care unit (ICU). We evaluated the effects of adjunctive administration of Remifentanil, a short-acting drug, in deeply sedated patient on variation of Bispectral Index (BIS) during a fiberoptic bronchoscopy. A prospective, randomized, blinded, placebo-controlled study was conducted in 18-bed ICU. Patients needing a tracheal fibroscopy under deep sedation (midazolam (0.1 mg/kg per hour) fentanyl (4 μg/kg per hour)) and neuromuscular blocking (atracurium 0.5 mg/kg) were included in the study. A continuous monitoring of BIS, arterial pressure, and heart rate were realized before, during, and after the fiberoptic exam. An adjunctive continuous placebo or Remifentanil infusion was started just before the fiberoptic exam with a target effect-site concentration of 4 ng/ml using a Base Primea pump. Mean arterial pressure and heart rates were comparable between the placebo and Remifentanil groups at all times of the procedure. We did not observe differences in the variation of BIS values between the two groups during procedure. We described no change in BIS values relative to the placebo group in this population. In deeply sedated and paralyzed patients, receiving analgesic support based on a scale score an additional administration of short-acting analgesic drug, such as Remifentanil, seems not to be necessary for acute pain control. NCT00162591.


Furosemide pharmacodynamics: effect of respiratory and acid-base disturbances.
The aims of this study were to investigate the effect of changes in arterial blood gases and pH on furosemide pharmacodynamics and kinetics. Five groups of conscious rabbits were used: a control group breathing air with normoxia and normocarbia; a second group with hypercapnia and respiratory acidosis; a third with hypoxemia; a fourth with hypercapnia and respiratory acidosis combined with hypoxemia (HCHO); and the fifth group with metabolic acidosis. All experimental conditions, except hypoxemia, increased sodium tubular reabsorption and therefore, decreased urinary excretion of sodium. Renal blood flow was decreased by HCHO and metabolic acidosis. In response to 5 mg/kv i.v. of furosemide, natriuresis and diuresis were decreased by an average of 44% in animals with HCHO (P less than .05). The kinetics of furosemide were not affected by any of the experimental conditions except HCHO, in which the renal clearance of furosemide was reduced from 7.5 +/- 1.4 ml/min/kg (controls) to 2.7 +/- 0.7 ml/min/kg (P less than .05). The reduction in renal clearance of furosemide was associated with a decrease in urinary excretion of sodium (P less than .05). The reduction in renal clearance of furosemide was probably secondary to the decrease in renal blood flow and an increase in furosemide tubular reabsorption. Finally, HCHO did not decrease plasma volume, suggesting that the reduction in renal blood flow was secondary to blood flow distribution. In conclusion, only hypercapnia and respiratory acidosis combined with hypoxemia decreases the natriuretic and diuretic effect of furosemide.


Three Giants in the Cradle of Reproductive Medicine; Reproduction Theories of the Seventeenth Century as Discerned by Pregnancy Portraiture in the Oeuvre of Jan Vermeer.
Portraits of pregnant women are rare in Catholic Renaissance art. In seventeenth-century Holland, the Catholic rule of Spain had been thrown off and a Protestant Calvinistic republic emerged, freeing Dutch artists to choose an unorthodox subject matter for their paintings. The Golden Age of Holland was characterized by extreme wealth, originating from overseas trade, which fostered a marked interest in philosophy, science, medicine, as well as art. Despite this, portraiture of pregnancy remained uncommon. An exception to this rule was Jan Vermeer of Delft, who lived during the zenith of this era. Jan Vermeer painted fewer than 40 pictures, fathered 15 children, and died bankrupt and little appreciated at the age of 43. Vermeer confined himself almost entirely to images of women in various domestic situations, including three figures of pregnant women. In this framework, pregnancy could be viewed as an icon for fidelity and conformism to social expectations. In this paper we investigate the roots of this unusual icon in Vermeer's oeuvre, and suggest that the use of pregnancy in his paintings could have been inspired by his Delft-resident contemporaries Antony van Leeuwenhoek and Reinier de Graaf, fathers of well-known and opposing theories of reproduction. These eminent scientists and Vermeer's pregnant wife, who frequently served as his model, might have made pregnancy less mysterious and more realistic to the painter.


Prickly Pear Cacti (Opuntia spp.) Cladodes as a Functional Ingredient for Hyperglycemia Management: A Brief Narrative Review.
The worldwide prevalence of obesity is increasing along with its comorbidities, including type 2 diabetes mellitus (T2DM). From a pathophysiological perspective, T2DM arises as a consequence of insulin resistance and pancreatic β-cell dysfunction, which together induce chronic hyperglycemia. The pharmacological treatment of T2DM specifically focuses on its management, rather than remission, with a lack of pharmacological agents to prevent the onset of the disease. Considering the role of unhealthy dietary patterns on the development of T2DM, identifying novel food ingredients and bioactive substances may provide new avenues by which to address the T2DM epidemic. In this brief review, we have summarized the latest findings on the consumption of the prickly pear (PP; Opuntia spp.) cladode as a potential nutritional tool for the management of hyperglycemia. The consumption of prickly pear cladodes was reported to exert hypoglycemic effects, making it a potential cost-effective nutritional intervention for the management of T2DM. Several studies have demonstrated that the consumption of prickly pear cladodes and the related products reduced post-prandial glucose levels. The cladodes' high fiber content may be implicated in improving glycemic control, by affecting glucose absorption and effectively slowing its release into the blood circulation. Given these potential hypoglycemic effects, prickly pear cladodes may represent a potential functional food ingredient to improve glycemic control and counter the negative metabolic effects of the modern Western diet. Nonetheless, in consideration of the lack of evidence on the chronic effects of the prickly pear cladode, future research aimed at evaluating its long-term effects on glycemic control is warranted.


Prolonged life-threatening hypoglycaemia following dose escalation of octreotide LAR in a patient with malignant polysecreting pancreatic neuroendocrine tumour.
This paper details the case of a 77-year-old male with refractory hypoglycaemia due to inoperable metastatic pancreatic neuroendocrine tumour (pNET) co-secreting insulin and gastrin. Multiple medical therapies were trialled with limited success, and we describe the complications experienced by our patient. Somatostatin analogues can ameliorate hypoglycaemia and may have tumour-stabilising effects; however, in our case resulted in paradoxical worsening of hypoglycaemia. This rendered our patient hospital dependent for glycaemic support including continuous dextrose infusion. Although this is a reported adverse effect with initiation of therapy, we describe successful initiation of short-acting octreotide as an inpatient followed by commencement of long-acting octreotide. Hypoglycaemic collapse occurred only after dose titration of long-acting octreotide. We outline the pitfalls of somatostatin analogue therapy and the mechanisms that may contribute to worsening hypoglycaemia. This rare side effect cannot be reliably predicted, necessitating close supervision and glucose monitoring during therapy. Our patient achieved disease stabilisation and gradual resolution of hypoglycaemia with peptide receptor radionuclide therapy (PRRT), an emerging therapeutic option for metastatic neuroendocrine tumours with high efficacy and low toxicity. We present a brief but comprehensive discussion of currently available and novel therapies for insulin secreting pNETs. Hypoglycaemia due to malignant insulin secreting pNET is frequently severe and may be life-threatening despite supportive therapies.Octreotide can ameliorate hypoglycaemia, and may have anti-proliferative and tumour-stabilising effects in malignant pNETs that are surgically unresectable.Paradoxical worsening of hypoglycaemia may occur with octreotide initiation and dose titration, necessitating close supervision and glucose monitoring.PRRT is emerging as a therapeutic option with high efficacy and low toxicity.


Internal- vs External-Connection Single Implants: A Retrospective Study in an Italian Population Treated by Certified Prosthodontists.
The design of an implant connection that allows prosthetic suprastructures to be attached to implants has long been debated in the dental literature. The goal of this retrospective study was to evaluate the 5-year clinical results for a large number of single implants restored by certified prosthodontists in an attempt to establish whether different clinical outcomes could be detected for external- or internal-connection implants. All single implants with internal or external connections inserted in 27 private dental practices from January 1, 2003 to December 31, 2007 were evaluated. An initial statistical analysis was performed to describe the sample population at baseline and then to compare the two types of implant-abutment connection configurations and their clinical outcomes. All data were statistically analyzed with STATA12 (StataCorp). Twenty-eight of the 85 active members of the Italian Academy of Prosthetic Dentistry (AIOP) participated in this study. The sample included 1,159 patients and 2,010 implants. Of the implants, 75 were dropped because there was no information about follow-up. Of the remaining implants, 1,431 (74.0%) were followed for at least 5 years, and 332 implants (17.2%) were followed for more than 8 years. Nearly 99% (98.9%) of the implants survived. The difference between the survival frequencies of the two types of implant-abutment connection configurations was not significant for each negative event (log-rank test, P > .05). There was no difference between the two types of implants regarding restoration fracture, implant screw loosening, and peri-implant disease. Within the limitations of this study, it can be suggested that there is no difference in clinical outcomes of single restorations joined to internal- or external-connection implants.