Post-transplantation hepatocellular carcinoma recurrence: Patterns and relation between vascularity and differentiation degree.
To evaluate the relationship between hepatocellular carcinoma (HCC) vascularity and grade; to describe patterns and vascular/histopathological variations of post-transplantation recurrence. This retrospective study included 165 patients (143 men, 22 women; median age 56.8 years, range 28-70.4 years) transplanted for HCC who had a follow-up period longer than 2 mo. Pre-transplantation dynamic computed tomography or magnetic resonance examinations were retrospectively reviewed, classifying HCC imaging enhancement pattern into hypervascular and hypovascular based on presence of wash-in during arterial phase. All pathologic reports of the explanted livers were reviewed, collecting data about HCC differentiation degree. The association between imaging vascular pattern and pathological grade was estimated using the Fisher exact test. All follow-up clinical and imaging data were reviewed for evidence of recurrence. Recurrence rate was calculated and imaging features of recurrent tumor were collected, classifying early and late recurrences based on timing (< or ≥ 2 years after transplantation) and intrahepatic, extrahepatic and both intrahepatic and extrahepatic recurrences based on location. All intrahepatic recurrences were classified as hypervascular or hypovascular and the differentiation degree was collected where available. The presence of variations in imaging enhancement pattern and pathological grade between the primary tumor and the intrahepatic recurrence was evaluated and the association between imaging and histopatholgical variations was estimated by using the χ(2) test. Of the 163 patients with imaging evidence of viable tumor, 156 (95.7%) had hypervascular and 7 (4.3%) hypovascular HCC. Among the 125 patients with evidence of viable tumor in the explanted liver, 19 (15.2%) had grade 1, 56 (44.8%) grade 2, 40 (32%) grade 3 and 4 (3.2%) grade 4 HCC, while the differentiation degree was not assessable for 6 patients (4.8%). A significant association was found between imaging vascularity and pathological grade (P = 0.035). Post-transplantation recurrence rate was 14.55% (24/165). All recurrences occurred in patients who had a hypervascular primary tumor. Three patients (12.5%) experienced late recurrence; the location of the first recurrence was extrahepatic in 14 patients (58.3%), intrahepatic in 7 patients (29.2%) and both intrahepatic and extrahepatic in 3 patients (12.5%). Two patients had a variation in imaging characteristics between the primary HCC (hypervascular) and the intrahepatic recurrent HCC (hypovascular), while 1 patient had a variation of histopathological characteristics (from moderate to poor differentiation), however no association was found between imaging and histopathological variations. A correlation was found between HCC grade and vascularity; some degree of variability may exist between the primary and the recurrence imaging/histopathological characteristics, apparently not correlated.


Psychiatry, compensation and rehabilitation.
In December 1984, the Victorian Government published its proposals for the reform of workers' compensation legislation. Many groups have expressed concern about the proposed changes. The literature contains studies of heterogeneous groups of patients from different countries with very different medical and legal systems. Assessment of the psychological outcome of patients with compensable injuries has not usually included the assessment of confounding variables such as other injuries, inter- and intrapersonal problems, and social circumstances. No firm conclusions can be reached about the relative benefits and disadvantages of lump sum and periodic payments from a psychological perspective.


[Splenic lymphoid structures reorganization in gerbils after space flight].
To assess the effects of weightlessness on the organism of mammals, splenic lymphoid tissue was studied in 10 intact Mongolian gerbils and in 10 animals after a 12-days-long spaceflight. In experimental animals the body mass and splenic, thymic and adrenal mass indexes were sharply reduced. In the spleen, the red pulp area was significantly increased, there appeared the areas of fibrous tissue proliferation, and extensive hemorrhage. Lymphoid nodules with germinal centers disappeared, while periarterial lymphoid sheaths were depleted. Quantitative analysis of cellular content of the lymphoid structures of the spleen showed dramatic activation of cell destruction, lymphocytopoiesis suppression with the disappearance of mitotically dividing cells and a decrease in the number of blast cells. The absence of plasma cells indicates the suppression of the processes of the immunocytopoiesis. The complex of changes noted suggests the decrease of immunological barrier of lymphoid tissue in the spleen and in the whole organism, in gerbils under the conditions of weightlessness in the spaceflight.


Thresholds of Toxicological Concern - Setting a threshold for testing below which there is little concern.
Low dose, low risk; very low dose, no real risk. Setting a pragmatic threshold below which concerns become negligible is the purpose of thresholds of toxicological concern (TTC). The idea is that such threshold values do not need to be established for each and every chemical based on experimental data, but that by analyzing the distribution of lowest or no-effect doses of many chemicals, a TTC can be defined - typically using the 5th percentile of this distribution and lowering it by an uncertainty factor of, e.g., 100. In doing so, TTC aims to compare exposure information (dose) with a threshold below which any hazard manifestation is very unlikely to occur. The history and current developments of this concept are reviewed and the application of TTC for different regulated products and their hazards is discussed. TTC lends itself as a pragmatic filter to deprioritize testing needs whenever real-life exposures are much lower than levels where hazard manifestation would be expected, a situation that is called "negligible exposure" in the REACH legislation, though the TTC concept has not been fully incorporated in its implementation (yet). Other areas and regulations - especially in the food sector and for pharmaceutical impurities - are more proactive. Large, curated databases on toxic effects of chemicals provide us with the opportunity to set TTC for many hazards and substance classes and thus offer a precautionary second tier for risk assessments if hazard cannot be excluded. This allows focusing testing efforts better on relevant exposures to chemicals.


The efficacy of Antipyretic Analgesics administration intravenously for Preventing Rocuronium-Associated Pain/Withdrawal Response: a systematic review and meta-analysis.
Rocuronium-associated injection pain/withdrawal response (RAIPWR) was non-ideal but occurred frequently when injection intravenously during anesthesia induction. Many studies had reported that pretreating with antipyretic analgesics (AAs) could reduce the occurrence of RAIPWR, but there was no consensus yet. Therefore, this meta-analysis was designed to systematically evaluate the benefits of AAs on RAIPWR in patients. PubMed, Cochrane Library, Ovid, EMbase, Chinese National Knowledge Infrastructure (CNKI), Wan Fang Data were searched by January 1st 2019 for randomized controlled trials (RCTs) applying AAs to alleviate RAIPWR in patients who underwent elective surgery under general anesthesia. Two investigators assessed quality of RCTs and extracted data respectively and the meta-analysis was carried on Revman 5.3 software. Moreover, we compared AAs in pros and cons directly with lidocaine, the most reported medicine to prevent RAIPWR. Data were analyzed from 9 RCTs totaling 819 patients. The results of Meta-analysis showed that compared to the control group, pretreating with AAs could prevent the total occurrence of RAIPWR [Risk ratio (RR), 0.52; 95% confidence interval (CI), 0.42 to 0.66; P < 0.0001], and took effect on moderate (RR, 0.56; 95%CI, 0.43 to 0.73; P < 0.0001) and severe RAIPWR (RR = 0.14; 95%CI, 0.08 to 0.24; P < 0.00001). When compared to lidocaine, the preventive effect was not so excellent as the latter but injection pain induced by prophylactic occurred less. The currently available evidence suggested that pretreating with AAs intravenously could alleviate RAIPWR. PROSPERO CRD42019129776.


Serum glypican 4 level in obese children and its relation to degree of obesity.
Previous adult studies have demonstrated associations of serum glypican 4 (Gpc4) and obesity parameters and insulin sensitivity. However, an association of serum Gpc4 and glucose metabolism remains contradictory. Study of serum Gpc4 in obese children has not been conducted. We aimed to determine serum Gpc4 levels in obese children with various degrees of obesity. Up to 370 overweight and obese children, aged 6-18 years were enrolled in this cross-sectional study. Oral glucose tolerance test (OGTT) was performed with fasting serum Gpc4, lipid profiles, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) measured. Insulin sensitivity and β-cell function indices were calculated from plasma glucose and serum insulin levels derived from the OGTT. Bioelectrical impedance analysis was performed for body fat determination. Comparisons of serum Gpc4 levels among the groups of children with various degrees of obesity were performed. Serum Gpc4 levels progressively increased in children with increasing body mass index standard deviation score (BMI SDS) tertiles [median (interquartile range, IQR): 2.3 (1.8, 3.2), 2.6 (1.9, 3.4) and 3.2 (2.4, 3.8) μg/L, P<.001]. There were no differences in serum Gpc4 levels among children in the different glucose metabolism categories. Log serum Gpc4 levels were positively correlated with SDSs of weight and BMI, cholesterol, AST and ALT. No associations of log serum Gpc4 and insulin sensitivity and β-cell function indices were demonstrated. Serum Gpc4 levels were increased with increasing degrees of obesity. There were no differences in serum Gpc4 levels among glucose metabolism categories.


The brain microenvironment mediates resistance in luminal breast cancer to PI3K inhibition through HER3 activation.
Although targeted therapies are often effective systemically, they fail to adequately control brain metastases. In preclinical models of breast cancer that faithfully recapitulate the disparate clinical responses in these microenvironments, we observed that brain metastases evade phosphatidylinositide 3-kinase (PI3K) inhibition despite drug accumulation in the brain lesions. In comparison to extracranial disease, we observed increased HER3 expression and phosphorylation in brain lesions. HER3 blockade overcame the resistance of HER2-amplified and/or PIK3CA-mutant breast cancer brain metastases to PI3K inhibitors, resulting in marked tumor growth delay and improvement in mouse survival. These data provide a mechanistic basis for therapeutic resistance in the brain microenvironment and identify translatable treatment strategies for HER2-amplified and/or PIK3CA-mutant breast cancer brain metastases.


An emerging occupational rehabilitation system in the People's Republic of China.
China has become a major economic influence in Asia and globally. The country is in the position to further develop its workers' insurance and compensation system. This paper aims to introduce the existing workers' compensation policies, explain how these policies guide the operation of the occupational rehabilitation system for injured workers, and suggest ways to further develop an effective and sustainable system for the country. Major government policies and initiative documents and existing literature on occupational rehabilitation were critically reviewed. Shortfalls in our current system were identified and potential further development regimes were propose. Since 2004, China has implemented its national policy on providing timely and comprehensive rehabilitation and return-to-work interventions for workers who are injured at work. The three-tier medical and occupational rehabilitation system appears effective for enabling injured workers to access these services. Such a system is regarded as the most optimal for the country in spearheading the development of quality occupational rehabilitation services, and at the same time incorporating the existing expertise in acute medical care and rehabilitation within the public medical and health system. Problems encountered in the system can be classified under the culture, system and competence building aspects. The future workers' insurance and compensation system can probably put more emphasis on using bio-psychosocial and work disability prevention models in guiding its service development and delivery. Efforts need to be placed on building the competence of professionals in the system who provide services for injured workers. The empowerment of important stakeholders in the workers' insurance and compensation system and their inclusion in the planning of service delivery are crucial for developing a sustainable and effective system for China.


[Phospholipids in blood serum and the differential diagnosis of chronic hepatitis B and the cirrhotic stage of the disease].
The study was carried out for the purpose of determining the diagnostic criteria applicable to the cirrhotic stage of chronic viral B-hepatitis (CVBH) by using the data of the phosphor-lipid spectrum of the blood serum. The phosphor-lipid spectrum of the blood serum was examined in 70 patients with CVBH and in 20 patients with small-nodular hepatic cirrhosis. Reliably lower contents of lysophosphatidyl serine (LPS), of lysophosphatidyl ethanolamine (LFE) and of phosphatidyl ethanolamine (PE) were found in patients with CVBH at the cirrhotic stage as compared to patients with CVBH, who did and did not abuse the alcohol. However, the latters had a higher level of the relative content of phosphatidyl choline (PC) as compared to the controls.


[Bonds and practices of care: correlations between health policies and forms of subjectivation].
This paper analyzes the bond as a strategy for practices of care in public health policies. The analysis was structured around the Foucauldian tools of truth and forms of government that constitute policies of subjectivation. The discussion surrounds the correlation between the National Policy of Primary Health Care and other documents of public policies as part of this approach that makes it possible to consider the articulation of the bond with practices of care. This reflection has considered the bond as a focus of experience constituted by the correlation between formation of knowledge, normativity of behaviors and forms of subjectivation with regard to a pragmatic of the subjects in their different modalities of relationship with themselves.


Assembly of Ruminococcus flavefaciens cellulosome revealed by structures of two cohesin-dockerin complexes.
Cellulosomes are sophisticated multi-enzymatic nanomachines produced by anaerobes to effectively deconstruct plant structural carbohydrates. Cellulosome assembly involves the binding of enzyme-borne dockerins (Doc) to repeated cohesin (Coh) modules located in a non-catalytic scaffoldin. Docs appended to cellulosomal enzymes generally present two similar Coh-binding interfaces supporting a dual-binding mode, which may confer increased positional adjustment of the different complex components. Ruminococcus flavefaciens' cellulosome is assembled from a repertoire of 223 Doc-containing proteins classified into 6 groups. Recent studies revealed that Docs of groups 3 and 6 are recruited to the cellulosome via a single-binding mode mechanism with an adaptor scaffoldin. To investigate the extent to which the single-binding mode contributes to the assembly of R. flavefaciens cellulosome, the structures of two group 1 Docs bound to Cohs of primary (ScaA) and adaptor (ScaB) scaffoldins were solved. The data revealed that group 1 Docs display a conserved mechanism of Coh recognition involving a single-binding mode. Therefore, in contrast to all cellulosomes described to date, the assembly of R. flavefaciens cellulosome involves single but not dual-binding mode Docs. Thus, this work reveals a novel mechanism of cellulosome assembly and challenges the ubiquitous implication of the dual-binding mode in the acquisition of cellulosome flexibility.


Altered thyroid status modulates portal pressure in normal rats.
Disturbances in thyroid function in humans and experimental animal models have been associated with alterations in liver function and portal circulation. We have previously shown that hypothyroidism can significantly reduce portal pressure in portal vein ligated rats as well as inhibit the development of cirrhosis and fulminant hepatic failure following toxic liver injury. The aim of this study was to determine the effects of increased and decreased thyroid function on portal pressure in rats with normal liver histology and portal circulation. Three groups of 12 Wistar rats each were studied over a 30 day period: euthyroid (Group 1), hyperthyroid (Group 2) and hypothyroid (Group 3). Hyperthyroidism was induced by subcutaneous injection of triiodothyronine (400 microg/100g body weight) every ten days during the study period. Hypothyroidism was induced by methimazole (0.04% in drinking water) from 2 weeks prior to and throughout the 30 day study. Serum triiodothyronine (T3) and thyroid stimulating hormone (TSH) levels were determined to confirm the induction of hyper- and hypothyroidism. Portal pressure was assessed by direct catheterization of the portal vein prior to sacrifice. Indirect confirmation of changes in portal circulation was obtained by determining splenic weight at the time of sacrificing the animals. Animals were sacrificed at 10 day intervals throughout the 30 day study. Triiodothyronine treated rats were hyperthyroid compared to controls, with an elevation in serum T3 levels (3.8+/-0.9 mmol/L vs 1.3+/-0.4 mmol/L, p<0.05). In rats treated with methimazole, hypothyroidism was confirmed by a 7-fold increase in serum TSH compared to controls (1.8+/-0.4 vs 0.24+/-0.04 mmol/L, p<0.01). Portal pressure was significantly higher in the triiodothyronine treated rats compared to controls (12.8+/-1.7 and 9.6+/-0.75 cm H2O, p<0.001). Splenic weights in hyperthyroid rats were significantly higher than in controls (579+/-44 vs 478+/-46 mg, p<0.01). Portal pressure was significantly lower in the methimazole treated group compared to the control group (8.13+/-0.68 vs 9.6+/-0.75 cm H2O, p<0.01) as were splenic weights (400+/-33 vs 478+/-46 mg, p<0.01). These studies demonstrate that disturbed thyroid function exerts significant hemodynamic effects on the portal circulation in normal rats and complements results from previous similar studies in cirrhotic animals.


Recognition of DNA modified by antitumor cisplatin by "latent" and "active" protein p53.
Tumor suppressor protein p53 possesses two DNA-binding sites. One that is located within its core domain is responsible for sequence-specific DNA binding of the protein, non-specific binding to internal segments of single- or double-stranded DNA, and to certain kinds of non-B DNA structures. The other that is contained in the C-terminus of the protein binds to damaged DNA. Binding of active, latent, and in vitro-activated p53 protein to DNA fragments modified by antitumor cisplatin was studied using electrophoretic mobility shift assay in agarose gels and immunoblotting analysis. We found that both latent and active p53 forms bound to random sequences of DNA globally modified by cisplatin with a higher affinity than to unmodified DNA. Interestingly, the latent form exhibited a more pronounced selectivity for platinated DNA than the active p53. Consistently with this observation, the preference of the latent form for platinated DNA decreased as a consequence of the activation of latent p53 by phosphorylation at the protein kinase C site within its C-terminus or by binding of the monoclonal antibody Bp53-10.1. Competition experiments involving a 20-bp consensus sequence of p53 suggested that the p53 core domain was a primary binding site of the active p53 when it bound to DNA fragments lacking consensus sequence, but modified by cisplatin. In addition, the latent protein was found to selectively interact with DNA modified by cisplatin probably via its C-terminus.


Preferential adherence of cable-piliated burkholderia cepacia to respiratory epithelia of CF knockout mice and human cystic fibrosis lung explants.
The Burkholderia cepacia complex consists of at least five well-documented bacterial genomovars, each of which has been isolated from the sputum of different patients with cystic fibrosis (CF). Although the world-wide prevalence of this opportunist pathogen in CF patients is low (1-3%), 'epidemic' clusters occur in geographically isolated regions. Prevalence in some of these clusters is as high as 30-40%. The majority of CF B. cepacia isolates belong to genomovar III, but the relationship between genomovar and virulence has not yet been defined. Because the initial stage of infection involves bacterial binding to host tissues, the present study investigated differences in the binding of representative isolates of all five genomovars to fixed nasal sections of UNC cftr (-/-) and (+/+) mice and to human lung explants, biopsy and autopsy tissue of CF and non-CF patients. Binding was highest for isolates of genomovar III, subgroup RAPD type 2, but only if the isolates expressed the cable pili phenotype. Antibodies to the 22-kDa adhesin of cable pili virtually abolished binding. Binding occurred only to cftr (-/-) nasal sections or to CF lung sections and was negligible in cftr (+/+) or human non-CF, histologically normal lung sections. Unlike normal epithelia, the hyperplastic epithelia of CF bronchioles were enriched in cytokeratin 13, a 55-kDa protein that has previously been shown to act as a receptor in vitro for cable-piliated B. cepacia. These findings may help to explain the high transmissibility ofCbl-positive, genomovar III strains of B. cepacia among CF patients.


Human Mycotoxin Biomonitoring: Conclusive Remarks on Direct or Indirect Assessment of Urinary Deoxynivalenol.
Deoxynivalenol is one of the most ubiquitous mycotoxins in the Western diet through its presence in cereals and cereal products. A vast amount of studies indicate the worrying level of exposure to this toxin, while even high percentages of the population exceed the tolerable daily intake. To evaluate and assess dietary exposure, analysis of urinary levels of deoxynivalenol and its glucuronides has been proposed as a reliable methodology. An indirect preliminary method was used based on the cleavage of deoxynivalenol glucuronides through the use of enzymes (β-glucuronidase) and subsequent determination of "total deoxynivalenol" (sum of free and released mycotoxins by hydrolysis). Next, a direct procedure for quantification of deoxynivalenol-3-glucuronide and deoxynivalenol-15-glucuronide was developed. As deoxynivalenol glucuronides reference standards are not commercially available, the indirect method is widely applied. However, to not underestimate the total deoxynivalenol exposure in urine, the direct and indirect methodologies need to be compared. Urinary samples (n = 96) with a confirmed presence of deoxynivalenol and/or deoxynivalenol glucuronides were analysed using both approaches. The indirect method clarified that not all deoxynivalenol glucuronides were transformed to free deoxynivalenol during enzymatic treatment, causing an underestimation of total deoxynivalenol. This short communication concludes on the application of direct or indirect assessment of urinary deoxynivalenol.


Determination of Tear Lipid Film Thickness Based on a Reflected Placido Disk Tear Film Analyzer.
This study aims at determining the thickness of the tear lipid layer (LL) observed from a placido-disc-based tear film analyzer. We prospectively collected reflections of placido-disk LL images using a tear film analyzer (Keratograph® 5M, Oculus) from subjects with dry eye symptoms. The LL thickness (LLT) over the inferior half of the cornea was estimated with the use of interference color analysis and the preprocessing of images with and without ring segmentation were obtained and analyzed. Moreover, LLTs before and after 1 h of applying topical ointment (Duratears, Alcon) were compared to validate the estimation of LLT. Our results suggested that the tear LLT can be assessed using a placido-disk-based tear film analyzer and interference color analysis. We verified a high correlation between non-segmented and segmented LL images and estimated LLT increase after applying ointment. In addition, we concluded that LLT can be evaluated by direct interference analysis without segmentation preprocessing.


The plant is crucial: specific composition and function of the phyllosphere microbiome of indoor ornamentals.
The plant microbiome is a key determinant of plant health. Less is known about the phyllosphere microbiota and its driving factors in built environments. To study the variability of the microbiome in relation to plant genotype and climate under different controlled conditions, we investigated 14 phylogenetically diverse plant species grown in the greenhouses of the Botanical Garden in Graz (Austria). All investigated plants showed specific bacterial abundances of up to 10(6) CFU cm(-2) on their leaves. Bacterial diversity (H('): 2.4-7.9) and number of putative OTUs (461-2013) were strongly plant species dependent. Statistical analysis showed a significantly higher correlation of community composition to plant genotype in comparison to the ambient climatic variables. In addition to the microbiome structure, we studied the antagonistic potential towards the foliar pathogen Botrytis cinerea as functional indicator. A high proportion of isolates (up to 58%) were able to inhibit pathogen growth by production of volatile organic compounds (VOCs). Data of structure and function were linked: frequently isolated VOCs producers (e.g. Bacillus and Stenotrophomonas) were highly present in phyllosphere communities, which were dominated by members of Firmicutes This study indicates that indoor ornamentals feature a distinct, stable microbiota on leaves irrespective of the indoor climate.


Molecular genetic analysis of a cattle population to reconstitute the extinct Algarvia breed.
Decisions to initiate conservation programmes need to account for extant variability, diversity loss and cultural and economic aspects. Molecular markers were used to investigate if putative Algarvia animals could be identified for use as progenitors in a breeding programme to recover this nearly extinct breed. 46 individuals phenotypically representative of Algarvia cattle were genotyped for 27 microsatellite loci and compared with 11 Portuguese autochthonous and three imported breeds. Genetic distances and factorial correspondence analyses (FCA) were performed to investigate the relationship among Algarvia and related breeds. Assignment tests were done to identify representative individuals of the breed. Y chromosome and mtDNA analyses were used to further characterize Algarvia animals. Gene- and allelic-based conservation analyses were used to determine breed contributions to overall genetic diversity. Genetic distance and FCA results confirmed the close relationship between Algarvia and southern Portuguese breeds. Assignment tests without breed information classified 17 Algarvia animals in this cluster with a high probability (q > 0.95). With breed information, 30 cows and three bulls were identified (q > 0.95) that could be used to reconstitute the Algarvia breed. Molecular and morphological results were concordant. These animals showed intermediate levels of genetic diversity (MNA = 6.0 +/- 1.6, Rt = 5.7 +/- 1.4, Ho = 0.63 +/- 0.19 and He = 0.69 +/- 0.10) relative to other Portuguese breeds. Evidence of inbreeding was also detected (Fis = 0.083, P < 0.001). The four Algarvia bulls had Y-haplotypes H6Y2 and H11Y2, common in Portuguese cattle. The mtDNA composition showed prevalence of T3 matrilines and presence of the African-derived T1a haplogroup. This analysis confirmed the genetic proximity of Algarvia and Garvonesa breeds (Fst = 0.028, P > 0.05). Algarvia cattle provide an intermediate contribution (CB = 6.18, CW = -0.06 and D1 = 0.50) to the overall gene diversity of Portuguese cattle. Algarvia and seven other autochthonous breeds made no contribution to the overall allelic diversity. Molecular analyses complemented previous morphological findings to identify 33 animals that can be considered remnants of the Algarvia breed. Results of genetic diversity and conservation analyses provide objective information to establish a management program to reconstitute the Algarvia breed.


Pharmacological Investigation of CC-LAAO, an L-Amino Acid Oxidase from Cerastes cerastes Snake Venom.
Snake venom proteins, which are responsible for deadly snakebite envenomation, induce severe injuries including neurotoxicity, myotoxicity, cardiotoxicity, hemorrhage, and the disruption of blood homeostasis. Yet, many snake-venom proteins have been developed as potential drugs for treating human diseases due to their pharmacological effects. In this study, we evaluated the use of, an L-amino acid oxidase isolated from Cerastes cerastes snake venom CC-LAAO, as a potential anti-glioblastoma drug, by investigating its in vivo and in vitro pharmacological effects. Our results showed that acute exposure to CC-LAAO at 1 and 2.5 µg/mL does not induce significant toxicity on vital organs, as indicated by the murine blood parameters including aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH) activities, and creatinine levels. The histopathological examination demonstrated that only at high concentrations did CC-LAAO induce inflammation and necrosis in several organs of the test subjects. Interestingly, when tested on human glioblastoma U87 cells, CC-LAAO induced a dose-dependent apoptotic effect through the H2O2 generated during the enzymatic reaction. Taken altogether, our data indicated that low concentration of CC-LAAO may be safe and may have potential in the development of anti-glioblastoma agents.


5-Hydroxytryptamine(1F) receptors do not participate in vasoconstriction: lack of vasoconstriction to LY344864, a selective serotonin(1F) receptor agonist in rabbit saphenous vein.
Recently, several novel approaches to the treatment of migraine have been advanced, including selective 5-hydroxytryptamine (or serotonin) 1B/1D (5-HT(1B/1D)) receptor agonists such as sumatriptan and 5-HT(1F) receptor agonists such as LY344864. Many 5-HT(1B/1D) receptor agonists have been identified based on their ability to produce cerebral vascular contraction, whereas LY344864 was identified as an inhibitor of trigeminal nerve-mediated dural extravasation. In our study, several triptan derivatives were compared with LY344864 for their ability to contract the rabbit saphenous vein, a tissue used in the preclinical identification of sumatriptan-related agonists. Sumatriptan, zolmitriptan, rizatriptan, and naratriptan all contracted the rabbit saphenous vein from baseline tone, whereas LY344864 in concentrations up to 10(-4) M did not contract the rabbit saphenous vein. Furthermore, vascular contractions to sumatriptan were markedly augmented in the presence of prostaglandin F(2alpha) (PGF(2alpha)). However, even in the presence of PGF(2alpha) (3 x 10(-7) M), LY344864 did not contract the rabbit saphenous vein in concentrations well in excess of its 5-HT(1F) receptor affinity (pK(i) = 8.2). Only when concentrations exceeded those likely to activate 5-HT(1B) and 5-HT(1D) receptors (>10(-5) M) did modest contractile responses occur in the presence of PGF(2alpha). Use of these serotonergic agonists revealed a significant correlation between the contractile potency in the rabbit saphenous vein and the affinities of these agonists at 5-HT(1B) and 5-HT(1D) receptors, although contractile agonist potencies were not quantitatively similar to 5-HT(1B) or 5-HT(1D) receptor affinities. In contrast, no significant correlation existed between the contractile potencies of these serotonergic agonists in the rabbit saphenous vein and their affinity at 5-HT(1F) receptors. These data support the contention that activation of 5-HT(1F) receptors will not result in vascular contractile effects.


Clinical diagnosis and chemical confirmation of ciguatera fish poisoning in New South Wales, Australia.
Ciguatera fish poisoning is common in tropical and sub-tropical areas and larger fish (> 10 kg) are more susceptible to toxin accumulation with age. Although the coastal climate of northern New South Wales is considered sub-tropical, prior to 2014 there has only been 1 documented outbreak of ciguatera fish poisoning from fish caught in the region. During February and March 2014, 2 outbreaks of ciguatera fish poisoning involved 4 and 9 individuals, respectively, both following consumption of Spanish mackerel from northern New South Wales coastal waters (Evans Head and Scotts Head). Affected individuals suffered a combination of gastrointestinal and neurological symptoms requiring hospital treatment. At least 1 individual was symptomatic up to 7 months later. Liquid chromatography-tandem mass spectrometry detected the compound Pacific ciguatoxin-1B at levels up to 1.0 µg kg(-1) in fish tissue from both outbreaks. During April 2015, another outbreak of ciguatera fish poisoning was reported in 4 individuals. The fish implicated in the outbreak was caught further south than the 2014 outbreaks (South West Rocks). Fish tissue was unavailable for analysis; however, symptoms were consistent with ciguatera fish poisoning. To our knowledge, these cases are the southernmost confirmed sources of ciguatera fish poisoning in Australia. Educational outreach to the fishing community, in particular recreational fishers was undertaken after the Evans Head outbreak. This highlighted the outbreak, species of fish involved and the range of symptoms associated with ciguatera fish poisoning. Further assessment of the potential for ciguatoxins to occur in previously unaffected locations need to be considered in terms of food safety.


Unraveling the Role of Cytochrome P450 as a Key Regulator Lantipeptide Production in Streptomyces globisporus.
The aim of this study was to investigate the regulation of lantipeptide production in Streptomyces globisporus SP6C4, which produces the novel antifungal lantipeptides conprimycin and grisin, and to identify the role of cytochrome P450 (P450) in tis regulation. To investigate the regulation of lantipeptide production, we created gene deletion mutants, including ΔP450, ΔtsrD, ΔlanM, ΔP450ΔtsrD, and ΔP450ΔlanM. These mutants were characterized in terms of their morphology, sporulation, attachment, and antifungal activity against Fusarium oxysporum. The gene deletion mutants showed distinct characteristics compared to the wild-type strain. Among them, the ΔP450ΔlanM double mutant exhibited a recovery of antifungal activity against F. oxysporum, indicating that P450 plays a significant role in regulating lantipeptide production in S. globisporus SP6C4. Our findings highlight the significant role of P450 in the regulation of lantipeptide production and morphological processes in S. globisporus. The results suggest a potential link between P450-mediated metabolic pathways and the regulation of growth and secondary metabolism in SP6C4, thereby highlighting P450 as a putative target for the development of new antifungal agents.


Linezolid and Vancomycin Resistant Enterococci: A Therapeutic Problem.
Enterococci are recognized as opportunistic pathogens, as well as commensals in both humans and animals. They are an important cause of nosocomial infections, difficult to treat, as the organism have intrinsic and acquired resistance to many antibiotics. To isolate and identify clinically relevant Enterococcus up to species level from all the clinical samples processed in the microbiology laboratory and also to study their resistance pattern. A prospective study was carried out for a period of one year from May 2014 to April 2015 at the Department of Microbiology, Subharti Medical College, Meerut, Uttar Pradesh, India. A total of 200 isolates of Enterococcus species from 15342 clinical samples obtained from IPD/OPD patients irrespective of age, having suspicion of bacterial infection were processed in the microbiology laboratory. Identification was done with standard biochemical methods. Antibiotic susceptibility testing was done on Muller Hilton agar plate by Kirby Bauer disc diffusion method. High-Level Gentamycin Resistance (HLGR) and High-Level Streptomycin Resistance (HLSR) were further confirmed by Agar dilution method and Broth microdilution method. Vancomycin and linezolid resistance was further confirmed by Agar dilution method and MIC was calculated by using VITEK 2, Biomerieux. All methodology was followed as per Clinical and Laboratory Standards Institute (CLSI) M100-S 24, 2014 guidelines. E. faecalis (n=169, 84.5%) was the predominant species isolated, followed by E. faecium (n=27, 13.5%) and E. casseliflavus (n=4, 2%). A total of 25 (12.5%) isolates were HLGR, 13 (6.5%) isolates were HLSR and 62 (31%) isolates were HLGR+HLSR. Vancomycin resistance was found in 14 (7%) isolates of which 11 (78.5%) were Van A and 3 (21.4%) were Van B, detected phenotypically as per relative MIC of vancomycin and teicoplanin. Linezolid resistance was seen in 4 (2%) of isolates which were vancomycin as well as high-level gentamycin and high-level streptomycin resistant. Isolation of Enterococcus species resistant to most of the higher antibiotics like vancomycin and linezolid, with high prevalence of High-Level Aminoglycoside Resistance (HLAR), from hospitalized patients is a major concern as such isolates have limited or no therapeutic option.


Methyl methanesulfonate mutagenesis in bacteriophage T4.
MMS induces diverse rII mutations from a wild-type background in bacteriophage T4. About 56% are base pair substitutions, about 30% are frameshift mutations, and the remainder is a miscellaneous set of rapidly reverting or leaky mutants of unknown composition; but deletions were not detected. MMS-induced forward mutation is sharply reduced by the mutations px and y, which also reduce ultraviolet, photodynamic and gamma-ray mutagenesis and increase killing by all of these agents. Thus, many of the mutations arise via the T4 WXY system. The induction of G:C leads to A:T transitions was detected even in a px or y background using sensitive reversion tests, and the few forward rII mutations that were induced from this background also behaved like transition mutations. Thus, some MMS-induced mutations arise independently of the WXY system, perhaps as a result of the (rather weak) ability of MMS to alkylate the O6 position of guanine.


Approaches to interventional fluoroscopic dose curves.
Modern fluoroscopes used for image-based guidance in interventional procedures are complex X-ray machines, with advanced image acquisition and processing systems capable of automatically controlling numerous parameters based on defined protocol settings. This study evaluated and compared approaches to technique factor modulation and air kerma rates in response to simulated patient thickness variations for four state-of-the-art and one previous-generation interventional fluoroscopes. A polymethyl methacrylate (PMMA) phantom was used as a tissue surrogate for the purposes of determining fluoroscopic reference plane air kerma rates, kVp, mA, and variable copper filter thickness over a wide range of simulated tissue thicknesses. Data were acquired for each fluoroscopic and acquisition dose curve within each vendor's default abdomen or body imaging protocol. The data obtained indicated vendor- and model-specific variations in the approach to technique factor modulation and reference plane air kerma rates across a range of tissue thicknesses. However, in the imaging protocol evaluated, all of the state-of-the-art systems had relatively low air kerma rates in the fluoroscopic low-dose imaging mode as compared to the previous-generation unit. Each of the newest-generation systems also employ Cu filtration within the selected protocol in the acquisition mode of imaging; this is a substantial benefit, reducing the skin entrance dose to the patient in the highest dose-rate mode of fluoroscope operation. Some vendors have also enhanced the radiation output capabilities of their fluoroscopes which, under specific conditions, may be beneficial; however, these increased output capabilities also have the potential to lead to unnecessarily high dose rates. Understanding how fluoroscopic technique factors are modulated provides insight into the vendor-specific image acquisition approach and may provide opportunities to optimize the imaging protocols for clinical practice.


Towards the biodegradation pathway of fosfomycin.
Three functionalised propylphosphonic acids were synthesised to study C-P bond cleavage in R. huakuii PMY1. (R)-1-Hydroxy-2-oxopropylphosphonic acid [(R)-5] was prepared by chiral resolution of (±)-dimethyl 1-hydroxy-2-methylallyllphosphonate [(±)-12], followed by ozonolysis and deprotection. The N-(l-alanyl)-substituted (1R,2R)-2-amino-1-hydroxypropylphosphonic acid 10, a potential precursor for 2-oxopropylphosphonic acid (5) in cells, was obtained by coupling the aminophosphonic acid with benzotriazole-activated Z-l-alanine and hydrogenolytic deprotection. (1R*,2R*)-1,2-Dihydroxy-3,3,3-trifluoropropylphosphonic acid, a potential inhibitor of C-P bond cleavage after conversion into its 2-oxo derivative in the cell, was accessed from trifluoroacetaldehyde hydrate via hydroxypropanenitrile 21, which was silylated and reduced to the aldehyde (±)-23. Diastereoselective addition of diethyl trimethylsilyl phosphite furnished diastereomeric α-siloxyphosphonates. The less polar one was converted to the desired racemic phosphonic acid (±)-(1R*,2R*)-9 as its ammonium salt.


On the adjustment for covariates in genetic association analysis: a novel, simple principle to infer direct causal effects.
In genetic association studies, different complex phenotypes are often associated with the same marker. Such associations can be indicative of pleiotropy (i.e. common genetic causes), of indirect genetic effects via one of these phenotypes, or can be solely attributable to non-genetic/environmental links between the traits. To identify the phenotypes with the inducing genetic association, statistical methodology is needed that is able to distinguish between the different causes of the genetic associations. Here, we propose a simple, general adjustment principle that can be incorporated into many standard genetic association tests which are then able to infer whether an SNP has a direct biological influence on a given trait other than through the SNP's influence on another correlated phenotype. Using simulation studies, we show that, in the presence of a non-marker related link between phenotypes, standard association tests without the proposed adjustment can be biased. In contrast to that, the proposed methodology remains unbiased. Its achieved power levels are identical to those of standard adjustment methods, making the adjustment principle universally applicable in genetic association studies. The principle is illustrated by an application to three genome-wide association analyses.


Tissue-engineered bone from BMP-2-transduced stem cells derived from human fat.
Progenitor cells capable of induction into multiple mesenchymal lineages have been isolated from human liposuction aspirates. These cells, named processed lipoaspirate cells, have previously shown in vitro osteogenic capacity. The purpose of this study was to examine the in vivo bone induction capacity of bone morphogenetic protein-2 (BMP-2)-transduced processed lipoaspirate cells using adipose tissue from multiple harvest sites. Processed lipoaspirate cells extracted from human abdominal and buttock liposuction aspirates (n = 5) and from infrapatellar fat pads (n = 5) were placed in osteogenic media containing Dulbecco's Modified Eagle Medium with 10% fetal bovine serum supplemented with 50 muM ascorbic acid-2-phosphate and 10 mM beta-glycerol phosphate. Half of these cells were transfected with an adenovirus carrying the cDNA for bone morphogenetic protein-2 (adBMP-2). These transfected cells were then seeded onto collagen I matrices at a concentration of 2 x 10 cells/matrix and were placed into the hind limbs of severe combined immunodeficient mice (n = 10). Nontransfected processed lipoaspirate cells were placed in the contralateral limb as a control. After 6 weeks, specimens were analyzed by radiographs, densitometry, and hematoxylin and eosin and von Kossa staining. The average number of cells extracted from the abdominal/buttock lipoaspirates was 3.4 x 10 cells/100 ml fat aspirate and 5.5 x 10 cells per infrapatellar fat pad (average volume, 20.6 cc). All 10 BMP-2 transfected processed lipoaspirate constructs produced abundant radiographic and histologic bone. The bone was adequately mineralized and was beginning to establish a marrow cavity. There was no quantitative difference in bone production between harvest sites [mean, 2.0 +/- 0.1 aluminum units (knee) versus 2.1 +/- 0.1 aluminum units (abdomen/buttock); p = 0.14]. No bone was produced in the negative controls. Multipotential processed lipoaspirate cells can be extracted from adipose tissue harvested from liposuction aspirates or from the infrapatellar fat pad of the knee. Processed lipoaspirate cells can be transduced with the BMP-2 gene to produce abundant in vivo bone. These cells appear to be clinically useful for bone tissue engineering applications either as osteoprogenitor cells or as delivery vehicles for BMP-2.


Hydroxyl-substituted ladder polyethers via selective tandem epoxidation/cyclization sequence.
A new and highly selective method for the synthesis of hydroxyl-substituted tetrahydropyrans is described. This method utilizes titanium(IV) isopropoxide and diethyl tartrate to perform a diastereoselective epoxidation followed by in situ epoxide activation and highly selective endo-cyclization to form the desired tetrahydropyran ring. The HIJ ring fragment of the marine ladder polyether yessotoxin was synthesized using this two-stage tactic that proceeds with high efficiency and excellent regioselectivity.


Sleep bruxism and oral health-related quality of life in children: A systematic review.
Sleep bruxism (SB) is a masticatory muscle activity during sleep that can cause several consequences to the stomatognathic system. This systematic review investigated the impact of SB on oral health-related quality of life (OHRQoL) of 0- to 6-year-old children. Literature search was undertaken through PubMed/MEDLINE, LILACS, Scopus, TRIP, Livivo databases, and grey literature. The search was conducted with no publication year or language limits. Two reviewers independently selected the studies, extracted the data and assessed the risk of bias. The quality of evidence was assessed using GRADE. From 185 potentially eligible studies, three were included in the review. All studies were conducted in Brazil, published between 2015 and 2017, and used the B-ECOHIS instrument to evaluate OHRQoL. Two studies found no association between SB and OHRQoL, whereas one showed a significant negative impact of SB on the OHRQoL of children. SB was associated with respiratory problems, presence of tooth wear, dental caries, malocclusion as well as income and pacifier use. Risk of bias ranged from moderate to high, and the quality of evidence was judged as very low. The evidence is currently insufficient for definitive conclusions about the impact of SB on OHRQoL of children.


Coxiella burnetii replicates in Galleria mellonella hemocytes and transcriptome mapping reveals in vivo regulated genes.
Larvae of the greater wax moth (Galleria mellonella) are susceptible to infection with C. burnetii, an obligate intracellular bacterial pathogen. We show that bacteria are found in hemocytes after infection, and occupy vacuoles which are morphologically similar to Coxiella-containing vacuoles seen in infected mammalian phagocytes. We characterized the infection by transcriptome profiling of bacteria isolated from the hemocytes of infected larvae and identified 46 highly upregulated genes. The encoded proteins are predicted to be involved in translation, LPS biosynthesis, biotin synthesis, scavenging of reactive oxygen species, and included a T4SS effector and 30 hypothetical proteins. Some of these genes had previously been shown to be upregulated in buffalo green monkey (BGM) cells or in mice, whilst others appear to be regulated in a host-specific manner. Altogether, our results demonstrate the value of the G. mellonella model to study intracellular growth and identify potential virulence factors of C. burnetii.


Simple PCR assays improve the sensitivity of HIV-1 subtype B drug resistance testing and allow linking of resistance mutations.
The success of antiretroviral therapy is known to be compromised by drug-resistant HIV-1 at frequencies detectable by conventional bulk sequencing. Currently, there is a need to assess the clinical consequences of low-frequency drug resistant variants occurring below the detection limit of conventional genotyping. Sensitive detection of drug-resistant subpopulations, however, requires simple and practical methods for routine testing. We developed highly-sensitive and simple real-time PCR assays for nine key drug resistance mutations and show that these tests overcome substantial sequence heterogeneity in HIV-1 clinical specimens. We specifically used early wildtype virus samples from the pre-antiretroviral drug era to measure background reactivity and were able to define highly-specific screening cut-offs that are up to 67-fold more sensitive than conventional genotyping. We also demonstrate that sequencing the mutation-specific PCR products provided a direct and novel strategy to further detect and link associated resistance mutations, allowing easy identification of multi-drug-resistant variants. Resistance mutation associations revealed in mutation-specific amplicon sequences were verified by clonal sequencing. Combined, sensitive real-time PCR testing and mutation-specific amplicon sequencing provides a powerful and simple approach that allows for improved detection and evaluation of HIV-1 drug resistance mutations.


The Role of Culture and Religion on Sexual and Reproductive Health Indicators and Help-Seeking Attitudes amongst 1.5 Generation Migrants in Australia: A Quantitative Pilot Study.
In Australia, 1.5 generation migrants (those who migrated as children) often enter a new cultural and religious environment, with its own set of constructs of sexual and reproductive health (SRH), at a crucial time in their psychosexual development-puberty/adolescence. Therefore, 1.5 generation migrants may thus have to contend with constructions of SRH from at least two cultures which may be at conflict on the matter. This study was designed to investigate the role of culture and religion on sexual and reproductive health indicators and help-seeking amongst 1.5 generation migrants. An online survey was completed by 111 participants who answered questions about their cultural connectedness, religion, sexual and reproductive health and help-seeking. Kruskall-Wallis tests were used to analyse the data. There was no significant difference between ethnocultural groups or levels of cultural connectedness in relation to sexual and reproductive health help-seeking attitudes. The results do suggest differences between religious groups in regard to seeking help specifically from participants' parents. Notably, participants who reported having 'no religion' were more likely to seek help with sexual and reproductive health matters from their parent(s). Managing cross-cultural experiences is often noted in the extant literature as a barrier to sexual and reproductive health help-seeking. However, while cultural norms of migrants' country of origin can remain strong, it is religion that seems to have more of an impact on how 1.5 generation migrants seek help for SRH issues. The findings suggest that 1.5 generation migrants may not need to adapt their religious beliefs or practices, despite entering a new ethnocultural environment. Given that religion can play a role in the participants' sexual and reproductive health, religious organizations are well-placed to encourage young migrants to adopt help-seeking attitudes.


Intramuscular, intravenous and oral levetiracetam in dogs: safety and pharmacokinetics.
Intravenous (IV) levetiracetam (LEV) is available for humans for bridge therapy when the oral route is unavailable. We investigated the safety and pharmacokinetics of LEV administered intramuscularly (IM), IV, and orally to dogs. Six Hound dogs received 19.5-22.6 mg/kg of LEV IM, IV and orally with a wash-out period in between. All dogs received 500 mg LEV orally and 5 mL of 100 mg/mL LEV IM. Three dogs received 500 mg of LEV IV and three dogs received 250 mg LEV IV with 250 mg given perivascularly to approximate extravasation. Safety was assessed using a pain scale at time of IM administration and histopathological examination 24 h to 5 days after injection. Intravenous LEV half-life was 180 +/- 18 min. Bioavailability of IM LEV was 100%. Mean time to T(max) after IM was 40 +/- 16 min. The mean C(max) IM was 30.3 +/- 3 mug/mL compared to the C(0) of 37 +/- 5 mug/mL for IV. Mean inflammation score (0-4 scale) for IM LEV was 0.28 and for saline 0.62. Extravasation did not cause tissue damage. Parenteral LEV is well tolerated and appears safe following IM and IV injections in dogs. Parenteral LEV should be evaluated for use in dogs with epilepsy.


Tissue variant effects of heme inhibitors on the mouse cytochrome c oxidase gene expression and catalytic activity of the enzyme complex.
The in vivo effects of heme biosynthesis inhibitors, succinylacetone and CoCl2 on the cytochrome c oxidase (COX) gene expression and enzyme activity in different mouse tissues were investigated. Succinylacetone and CoCl2 showed tissue-specific differences in their ability to modulate heme aa3 content. A single dose of succinylacetone treatment for 8 h reduced the heme aa3 content of kidney mitochondria with no effect on the liver. CoCl2 treatment for 8 h, however, selectively affected the heme aa3 level in the liver. Reduced mitochondrial heme aa3 with both treatments was accompanied by approximately 50% reduced, mitochondrial genome-encoded COX I and II mRNAs and nuclear genome-encoded COX Vb mRNAs, but no change in COX IV mRNA level. Use of isolated mouse liver and brain mitochondrial systems showed a 50-80% reduction in mitochondrial transcription and translation rates in heme-depleted tissues. Blue native gel electrophoresis followed by immunoblot analysis showed that the complex from heme-depleted tissues contained a 30-50% reduction in levels of subunits I, IV, Vb and near normal levels of subunit VIc, indicating altered subunit content. Treatment of submitochondrial particles with protein kinase A and ATP resulted in partial dissociation of COX, suggesting a mechanistic basis for the reduced subunit content of the complex from heme-depleted tissues. Surprisingly, the enzyme from heme-depleted tissues showed twofold to fourfold higher turnover rates for cytochrome c oxidation, suggesting alterations in the kinetic characteristics of the enzyme following heme reduction. This is probably the first evidence that the tissue heme level regulates not only the mammalian COX gene expression, but also the catalytic activity of the enzyme, probably by affecting its stability.


Does the historical literature on encephalitis lethargica support a simple (direct) relationship with postencephalitic Parkinsonism?
This article and the subsequent one suggest that the currently accepted view of a simplistic (direct) relationship between encephalitis lethargica (EL) and postencephalitic Parkinsonism (PEP) is based on a incomplete evaluation of the epidemic period literature. In this article we provide a detailed analysis of the literature from the period that demonstrates that Parkinsonism was not initially part of acute EL symptomatology, that PEP was not typically the prevailing type of chronic EL and that oculogyric crises were never part of acute EL symptomatology and not initially associated with PEP. The second paper uses these finding, and also examines the clinical justifications for concluding that all patients with PEP had prior acute episodes of EL, to reevaluate the presumed direct etiologic relationship between EL and PEP.


Association of wake time and the onset of myocardial infarction. Triggers and mechanisms of myocardial infarction (TRIMM) pilot study. TRIMM Study Group.
A morning increase in onset of acute myocardial infarction (MI) has been documented, but its association with wake time and possible triggering events is unclear. The community-based, ongoing Triggers and Mechanisms of Myocardial Infarction (TRIMM) study was designed to investigate the factors associated with the transition from chronic coronary artery disease to acute MI. During the pilot phase in 1989, 224 consecutive hospitalized MI patients (176 men and 48 women, aged 60.3 +/- 9.2 years) of the prospectively defined Monitoring of Trends and Determinates of Cardiovascular Disease (MONICA) Augsburg MI register were interviewed 16.8 +/- 6.5 days after the event. The frequency of onset of MI was significantly higher (p less than 0.01) during the morning from 6 to 9 AM compared with other times of day. After adjustment for individual wake times, the peak of onset of MI was markedly sharper; the relative risk of MI during the 3-hour peak interval (the 3 hours after awakening) compared with other times of day increased from 1.8 (95% confidence interval, 1.3-2.4) to 2.4 (95% confidence interval, 1.8-3.1). Sixty-seven percent of the patients reported possible acute triggers of MI and/or unusual life events; among those, 52% reported stress or emotional upset. The TRIMM pilot study demonstrated the feasibility of a community-based study of possible triggering events of MI. The circadian variation of MI appears to result primarily from increased onset after awakening. This narrowing of the time frame of increased risk of MI should facilitate study of pathogenic mechanisms and aid in the design of more effective preventive regimens.


[Elderly women dealing with institutionalization].
This study focused on institutionalization of the elderly from the perspective of gender studies. The aim was to understand the effects of institutionalization on the lives of elderly women and their strategies to deal with this situation. The study used a qualitative methodology and was conducted in a long-term care facility for the elderly, with 110 individuals from Alto Uruguai, Rio Grande do Sul State, Brazil. Data collection used discussion groups with ten elderly women participating. Content analysis was used and identified three main categories: the nursing home as a total institution, gender, and strategies to deal with institutionalization. Half of the women had chosen to live in the nursing home, while the others complained about their confinement to what they considered "a dump for old people". The gender category permeated the women's lives, which included domestic activities in the "home" in order to pass the time. Strategies to deal with institutionalization included religious rituals, handicrafts, and walks.


An enzyme-responsive controlled release system based on a dual-functional peptide.
A new controlled release system was developed by loading a dual-functional peptide (DFP) on a mesoporous silica material. One-pot synthesis produced a DFP that was stimuli responsive, releasing a therapeutic peptide by protease cleavage. The design provides new steps towards smart biomaterials.


Noise stress changes mRNA expressions of corticotropin-releasing hormone, its receptors in amygdala, and anxiety-related behaviors.
Noise is a psychological, environmental stressor that activates limbic sites in the brain. Limbic sites such as the amygdala and the amygdaloid corticotropin-releasing hormone (CRH) system play an important role in integrating stress response. We investigated the association between noise exposures, CRH-related molecules in the amygdala, and behavioral alterations. In total 54 Sprague-Dawley rats were divided into the following three groups: Control (CON), acute noise exposure (ANE), and chronic noise exposure (CNE). The ANE group was exposed to 100 dB white noise only once in 4 h and the CNE group was exposed to the same for 4 h per day for 30 days. Expression profiles of CRH and its receptors CRH-R1 and CRH-R2 were analyzed by quantitative real-time polymerase chain reaction (qPCR). The same stress procedure was applied to the ANE and CNE groups for behavior testing. The anxiety responses of the animals after acute and chronic stress exposure were measured in the defensive withdrawal test. CNE upregulated CRH and CRH-R1 mRNA levels but downregulated CRH-R2 mRNA levels. ANE led to a decrease in both CRH-R1 and CRH-R2 expression. In the defensive withdrawal test, while the ANE increased, CNE reduced anxiety-like behaviors. The present study shows that the exposure of rats to white noise (100 dB) leads to behavioral alterations and molecule-specific changes in the CRH system. Behavioral alterations can be related to these molecular changes in the amygdala.


Extraocular rectus muscles of the rabbit contain alpha 2- and beta 2-adrenergic receptors.
Selective and non-selective adrenergic drugs were given to isolated extraocular rectus muscles (EOM) of rabbits to differentiate its postjunctional adrenergic receptors. Without precontraction none of the adrenergic agents had significant effects, but after cholinergic precontraction of the muscles a further contraction could be obtained with the non-selective agonists epinephrine, norepinephrine and isoproterenol, with the alpha 2-agonist B-HT 920, and with the beta 2-agonist terbutaline. No effect was seen from alpha 1- and beta 1-agonists. The presence of alpha 2- and beta 2-receptors was confirmed in experiments with selective and non-selective adrenergic antagonists. The lack of alpha 1-receptors in EOM and the absence of a hypertensive peak after topical alpha 2- and beta 2-agonists in vivo does not support the assumption that the initial rise in intraocular pressure (IOP) after topical epinephrine is produced by contraction of EOM.


Incidence of Echinococcus granulosus in Domestic Dogs in Palestine as Revealed by Copro-PCR.
Hydatidosis or echinococcosisis considered a neglected zoonotic disease despite its high burden in the livestock industry and the high risk of infection by humans in endemic areas. In a cross-sectional study we estimated the copro-Incidence and also genotyped Echinococcus granulosus isolates from domestic dogs using polymerase chain reaction (PCR). Medical archives in nine major hospitals in Palestine were reviewed to determine incidence of E. granulosus infection detected in humans during surgery. Faecal samples were collected from 93 domestic dogs in three districts with the highest number of human cases: Al-Khalil (Hebron), Tubas and Jenin. Genomic DNA was extracted from dog faecal samples and amplified by PCR targeting the repeat DNA sequence (EgG1 Hae III) followed by sequencing of five positive samples. Genotyping was determined by sequencing and BLAST searching of mitochondrial cytochrome c oxidase subunit (CO1). The incidence of E. granulosus infection detected in humans at surgery was 1.2 per 100,000 in the West Bank and 1.0 per 100,000 in Gaza Strip. Seventeen of 93 domestic dogs (18%) were positive, based upon comparison with the Echinococcus DNA control. The five sequenced samples were confirmed to be E. granulosus. Successfully genotyped sample belonged to E.granulosus sensu stricto (formerly G1-G3 complex, sheep strain). For domestic dogs, age group (13-24 months) and sex were identified as two risk factors for contracting E. granulosus. The study identified the high incidence of E. granulosus sensu stricto in dogs in Palestine.


Does cognitive performance map to categorical diagnoses of schizophrenia, schizoaffective disorder and bipolar disorder? A discriminant functions analysis.
Despite known overlaps in the pattern of cognitive impairments in individuals with bipolar disorder (BD), schizophrenia (SZ) and schizoaffective disorder (SZA), few studies have examined the extent to which cognitive performance validates traditional diagnostic boundaries in these groups. Individuals with SZ (n=49), schizoaffective disorder (n=33) and BD (n=35) completed a battery of cognitive tests measuring the domains of processing speed, immediate memory, semantic memory, learning, working memory, executive function and sustained attention. A discriminant functions analysis revealed a significant function comprising semantic memory, immediate memory and processing speed that maximally separated patients with SZ from those with BD. Initial classification scores on the basis of this function showed modest diagnostic accuracy, owing in part to the misclassification of SZA patients as having SZ. When SZA patients were removed from the model, a second cross-validated classifier yielded slightly improved diagnostic accuracy and a single function solution, of which semantic memory loaded most heavily. A cluster of non-executive cognitive processes appears to have some validity in mapping onto traditional nosological boundaries. However, since semantic memory performance was the primary driver of the discrimination between BD and SZ, it is possible that performance differences between the disorders in this cognitive domain in particular, index separate underlying aetiologies.


Androgen induces adaptation to oxidative stress in prostate cancer: implications for treatment with radiation therapy.
Radiation therapy is a standard treatment for prostate cancer (PC). The postulated mechanism of action for radiation therapy is the generation of reactive oxygen species (ROS). Adjuvant androgen deprivation (AD) therapy has been shown to confer a survival advantage over radiation alone in high-risk localized PC. However, the mechanism of this interaction is unclear. We hypothesize that androgens modify the radioresponsiveness of PC through the regulation of cellular oxidative homeostasis. Using androgen receptor (AR)(+) 22rv1 and AR(-) PC3 human PC cell lines, we demonstrated that testosterone increased basal reactive oxygen species (bROS) levels, resulting in dose-dependent activation of phospho-p38 and pAKT, and increased expression of clusterin, catalase, and manganese superoxide dismutase. Similar data were obtained in three human PC xenografts; WISH-PC14, WISH-PC23, and CWR22, growing in testosterone-supplemented or castrated SCID mice. These effects were reversible through AD or through incubation with a reducing agent. Moreover, testosterone increased the activity of catalase, superoxide dismutases, and glutathione reductase. Consequently, AD significantly facilitated the response of AR(+) cells to oxidative stress challenge. Thus, testosterone induces a preset cellular adaptation to radiation through the generation of elevated bROS, which is modified by AD. These findings provide a rational for combined hormonal and radiation therapy for localized PC.


Modeling of temperature-induced near-infrared and low-field time-domain nuclear magnetic resonance spectral variation: chemometric prediction of limonene and water content in spray-dried delivery systems.
The influence of temperature on near-infrared (NIR) and nuclear magnetic resonance (NMR) spectroscopy complicates the industrial applications of both spectroscopic methods. The focus of this study is to analyze and model the effect of temperature variation on NIR spectra and NMR relaxation data. Different multivariate methods were tested for constructing robust prediction models based on NIR and NMR data acquired at various temperatures. Data were acquired on model spray-dried limonene systems at five temperatures in the range from 20 degrees C to 60 degrees C and partial least squares (PLS) regression models were computed for limonene and water predictions. The predictive ability of the models computed on the NIR spectra (acquired at various temperatures) improved significantly when data were preprocessed using extended inverted signal correction (EISC). The average PLS regression prediction error was reduced to 0.2%, corresponding to 1.9% and 3.4% of the full range of limonene and water reference values, respectively. The removal of variation induced by temperature prior to calibration, by direct orthogonalization (DO), slightly enhanced the predictive ability of the models based on NMR data. Bilinear PLS models, with implicit inclusion of the temperature, enabled limonene and water predictions by NMR with an error of 0.3% (corresponding to 2.8% and 7.0% of the full range of limonene and water). For NMR, and in contrast to the NIR results, modeling the data using multi-way N-PLS improved the models' performance. N-PLS models, in which temperature was included as an extra variable, enabled more accurate prediction, especially for limonene (prediction error was reduced to 0.2%). Overall, this study proved that it is possible to develop models for limonene and water content prediction based on NIR and NMR data, independent of the measurement temperature.


2,3,7,8-Tetrachlorodibenzo-p-dioxin-induced activation of a protein tyrosine kinase, pp60src, in murine hepatic cytosol using a cell-free system.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) was found to activate protein kinases under cell- and nucleus-free conditions in isolated C57 mouse liver cytosol (100,000 x g supernatant). This action of TCDD was found to be aryl hydrocarbon receptor (AHR) dependent, concentration dependent, and inhibited by genistein, a tyrosine kinase inhibitor. The lowest concentration of TCDD to produce a statistically significant increase in protein phosphorylation was 10 pM. We also investigated the possibility that a protein kinase is physically associated with the cytosolic AHR complex. Kinase renaturation tests designed to detect reactivated protein kinases after electrophoresis in sodium dodecyl sulfate-polyacrylamide gels revealed the presence of a 60-kDa kinase in the washed immunoprecipitate obtained from liver cytosol using anti-AHR antibody (IgG) and protein A/G/agarose beads but not when a nonspecific IgG was used instead of anti-AHR antibody. The same 60-kDa band was present in an immunoprecipitate prepared in a similar manner from the same cytosol but with anti-heat shock protein 90 antibody (IgM). This 60-kDa kinase was found to be activated by TCDD treatment of whole cytosol from untreated mice. Moreover, pp60(src) immunoprecipitated from cytosol that had been previously treated with TCDD under cell-free conditions exhibited 2-fold more kinase activity than the equivalent preparation treated with a solvent control. Again, such an effect of TCDD could not be detected when a nonspecific IgG was used in place of an anti-pp60(src) antibody. Increased protein phosphorylation was observed after direct TCDD treatment of immunoprecipitates obtained using antibodies to AHR and pp60(src), respectively, but not when a nonspecific IgG was used for immunoprecipitation in either case. This observation is consistent with the idea that in cytosol, the AHR and pp60(src) coexist as part of a multimeric protein complex that can be specifically coimmunoprecipitated. These results provide evidence that (i) TCDD activates protein kinases in murine hepatic cytosol, (ii) a 60-kDa protein kinase is associated with the cytosolic form of the AHR complex, (iii) ligand binding directly activates this kinase because TCDD treatment of immunoprecipitated AHR complex results in increased protein kinase activity, and (iv) the AHR-associated protein kinase seems to be pp60(src) kinase. The current findings provide a clue to a potentially important mechanism by which TCDD can exert rapid, pleiotropic effects through the AHR-associated kinase to alter functions of many proteins through a cascade of protein phosphorylations.


Translesion synthesis mechanisms depend on the nature of DNA damage in UV-irradiated human cells.
Ultraviolet-induced 6-4 photoproducts (6-4PP) and cyclobutane pyrimidine dimers (CPD) can be tolerated by translesion DNA polymerases (TLS Pols) at stalled replication forks or by gap-filling. Here, we investigated the involvement of Polη, Rev1 and Rev3L (Polζ catalytic subunit) in the specific bypass of 6-4PP and CPD in repair-deficient XP-C human cells. We combined DNA fiber assay and novel methodologies for detection and quantification of single-stranded DNA (ssDNA) gaps on ongoing replication forks and postreplication repair (PRR) tracts in the human genome. We demonstrated that Rev3L, but not Rev1, is required for postreplicative gap-filling, while Polη and Rev1 are responsible for TLS at stalled replication forks. Moreover, specific photolyases were employed to show that in XP-C cells, CPD arrest replication forks, while 6-4PP are responsible for the generation of ssDNA gaps and PRR tracts. On the other hand, in the absence of Polη or Rev1, both types of lesion block replication forks progression. Altogether, the data directly show that, in the human genome, Polη and Rev1 bypass CPD and 6-4PP at replication forks, while only 6-4PP are also tolerated by a Polζ-dependent gap-filling mechanism, independent of S phase.


CsrA coordinates the expression of ribosome hibernation and anti-σ factor proteins.
Bacterial growth rate varies due to changing physiological signals and is fundamentally dependent on protein synthesis. Consequently, cells alter their transcription and translation machinery to optimize the capacity for protein production under varying conditions and growth rates. Our findings demonstrate that the post-transcriptional regulator CsrA in Escherichia coli controls the expression of genes that participate in these processes. During exponential growth, CsrA represses the expression of proteins that alter or inhibit RNA polymerase (RNAP) and ribosome activity, including the ribosome hibernation factors RMF, RaiA, YqjD, ElaB, YgaM, and SRA, as well as the anti-σ70 factor, Rsd. Upon entry into the stationary phase, RaiA, YqjD, ElaB, and SRA expression was derepressed and that of RMF, YgaM, and Rsd was activated in the presence of CsrA. This pattern of gene expression likely supports global protein expression during active growth and helps limit protein production to a basal level when nutrients are limited. In addition, we identified genes encoding the paralogous C-tail anchored inner membrane proteins YqjD and ElaB as robust, direct targets of CsrA-mediated translational repression. These proteins bind ribosomes and mediate their localization to the inner cell membrane, impacting a variety of processes including protein expression and membrane integrity. Previous studies found that YqjD overexpression inhibits cell growth, suggesting that appropriate regulation of YqjD expression might play a key role in cell viability. CsrA-mediated regulation of yqjD and ribosome hibernation factors reveals a new role for CsrA in appropriating cellular resources for optimum growth under varying conditions.IMPORTANCEThe Csr/Rsm system (carbon storage regulator or repressor of stationary phase metabolites) is a global post-transcriptional regulatory system that coordinates and responds to environmental cues and signals, facilitating the transition between active growth and stationary phase. Another key determinant of bacterial lifestyle decisions is the management of the cellular gene expression machinery. Here, we investigate the connection between these two processes in Escherichia coli. Disrupted regulation of the transcription and translation machinery impacts many cellular functions, including gene expression, growth, fitness, and stress resistance. Elucidating the role of the Csr system in controlling the activity of RNAP and ribosomes advances our understanding of mechanisms controlling bacterial growth. A more complete understanding of these processes could lead to the improvement of therapeutic strategies for recalcitrant infections.


TRANCE- and CD40 ligand-matured dendritic cells reveal MHC class I-restricted T cells specific for autologous tumor in late-stage ovarian cancer patients.
The use of mature dendritic cells (DCs) presenting tumor-associated antigens (TAAs) to trigger tumor-specific T cells in vivo or in vitro represents a promising approach for cancer immunotherapy. We hypothesized that tumor antigens, mostly unidentified, are present on ovarian tumor cells and that mature DCs could be used to generate tumor-specific responses in unprimed patients. We also sought to measure preexisting antitumor immunity in patients with advanced ovarian cancer. Autologous DCs from 10 patients with ovarian cancer were pulsed with killed autologous primary tumors as a source of TAAs. DCs were then cultured in the presence of tumor necrosis factor alpha + TRANCE (tumor necrosis factor-related activation-induced cytokine) to induce maturation. Mature TAA-pulsed DCs were used in vitro to stimulate tumor-specific peripheral blood T cells. TRANCE and CD40 ligand were effective at maturing DCs. T-cell lines were generated in vitro that were capable of secreting IFN-gamma in response to autologous tumor. These tumor-specific T cells were MHC class I restricted. The frequency of tumor-specific T cells in uncultured cells from malignant ascites fluid and peripheral blood was measured in the same patients. IFN-gamma-secreting tumor-specific T cells were demonstrated at baseline in uncultured T cells from some unvaccinated ovarian cancer patients; however, the T cells could not kill autologous tumor. These data demonstrate that mature DCs presenting tumor antigens from engulfed autologous tumors can be used to augment antitumor immunity in vitro in patients with epithelial ovarian cancer. The results support the feasibility of therapeutic vaccination of ovarian cancer patients.


Tissue-type plasminogen activator (t-PA) is stored in Weibel-Palade bodies in human endothelial cells both in vitro and in vivo.
Vascular endothelial cells are thought to be the main source of plasma tissue-type plasminogen activator (t-PA) and von Willebrand factor (VWF). Previous studies have suggested that both t-PA and VWF are acutely released in response to the same stimuli, both in cultured endothelial cells and in vivo. However, the subcellular storage compartment in endothelial cells has not been definitively established. We tested the hypothesis that t-PA is localized in Weibel-Palade (WP) bodies, the specialized endothelial storage granules for VWF. In cultured human umbilical vein endothelial cells (HUVECs), t-PA was expressed in a minority of cells and found in WP bodies by immunofluorescence. After up-regulation of t-PA synthesis either by vascular endothelial growth factor (VEGF) and retinoic acid or by sodium butyrate, there was a large increase in t-PA-positive cells. t-PA was exclusively located to WP bodies, an observation confirmed by immunoelectron microscopy. Incubation with histamine, forskolin, and epinephrine induced the rapid, coordinate release of both t-PA and VWF, consistent with a single storage compartment. In native human skeletal muscle, t-PA was expressed in endothelial cells from arterioles and venules, along with VWF. The 2 proteins were found to be colocalized in WP bodies by immunoelectron microscopy. These data indicate that t-PA and VWF are colocalized in WP bodies, both in HUVECs and in vivo. Release of both t-PA and VWF from the same storage pool likely accounts for the coordinate increase in the plasma level of the 2 proteins in response to numerous stimuli, such as physical activity, beta-adrenergic agents, and 1-deamino-8d-arginine vasopressin (DDAVP) among others.


Clinical and epidemiological aspects of hepatocellular carcinoma in Brazil.
The global hepatocellular carcinoma (HCC) incidence is widely variable, depending on geographic region and the prevalence of major risk factors. In Brazil, two large multicentre retrospective studies were performed to investigate clinical and epidemiological aspects of HCC. In the first study, performed in 1997, HCC was found in cirrhotic livers in 71% of cases. Chronic alcoholism was present in 36% of cases, chronic hepatitis B in 35% and hepatitis C in 25%. In a 2010 survey, cirrhosis was present in 98% of cases and HCV was the main aetiology (54%). Differences in HBV prevalence were found among regions. Selection of HCC treatment depends on tumour burden, liver function and performance status. Liver transplantation (LT) is the best available curative treatment for HCC in its early stage and with compromised liver function. After modifications in priority policy, the number of patients with early HCC submitted for LT has increased in the past 5 years in Brazil. Chemoembolization is the most common initial HCC therapy in early and intermediate stages of HCC in Brazil.


Development of the pyruvate kinase deficiency diary and pyruvate kinase deficiency impact assessment: Disease-specific assessments.
Currently recommended patient-reported outcome (PRO) measures for patients with pyruvate kinase (PK) deficiency are non-disease-specific. The PK Deficiency Diary (PKDD) and PK Deficiency Impact Assessment (PKDIA) were developed to be more targeted measures for capturing the symptoms and impacts of interest to this patient population. The instruments were developed based on concept elicitation interviews with 21 adults and modified based on 20 cognitive interviews. The domain structure and item concepts of the PKDD and PKDIA were compared with currently recommended measures, the EORTC QLQ-C30 and the SF-36v2®. The PKDD is a seven-item measure of the core signs and symptoms of PK deficiency. The PKDIA is a 14-item measure of the impacts of PK deficiency on patients' health-related quality of life (HRQoL). Minimal similarities were found between the new measures and the EORTC QLQ-C30 (eg, 43% of concepts were similar to the PKDD; 42% were similar to the PKDIA) and SF-36v2® (57% of concepts were similar to the PKDD; 17% were similar to the PKDIA). The PKDD and PKDIA fill a gap in the existing outcomes measurement strategy for PK deficiency. Future work includes psychometric evaluation of these newly developed measures.


A Phase II Trial to Assess the Safety and Efficacy of a Topical Repair Cream Containing Skin-identical Ceramide Complex in Patients with Contact Dermatitis.
Background: Contact dermatitis is a common skin condition observed by dermatologists, presenting a burden on healthcare systems. Recently, there has been a trend in producing skin-identical topical preparations for the repair of skin. However, there is a limited number of experimental studies to assess the safety and efficacy of this products. Objective: This study assessed the clinical efficacy and safety of a skin-identical ceramide complex cream (Dermalex Repair Contact Eczema; Omega Pharma, Nazareth, Belgium) in the treatment of contact dermatitis. Design: This was a Phase II, before-after trial. Setting: This study was conducted at the Center for Research and Training in Skin Diseases and Leprosy (CRTSDL) at Tehran University of Medical Sciences in Tehran, Iran. Participants: Fifteen patients with contact dermatitis (8 men and 7 women) between the ages of 25 and 62 years (median age: 36.4 years) were enrolled in this study. Measurements: Changes were assessed using six skin biophysical parameters (transepidermal water loss [TEWL], stratum corneum [SC] hydration, melanin index, erythema index, skin pH, and skin friction), Physician Global Assessment (PGA) score, and Three-Item Severity (TIS) score at baseline, Week 2, and Week 4 of the study. Results: Skin hydration and TIS showed a statistically significant improvement after treatment with study cream (p=0.023 and p=0.007, respectively). Although the reduction in TEWL was not significant, a slight decrease was observed at Week 4. Conclusions: The skin-identical ceramide complex cream improved contact dermatitis with a decrease in TIS and an increase in skin hydration, implying a repair of the skin barrier.


Tungsten-182 heterogeneity in modern ocean island basalts.
New tungsten isotope data for modern ocean island basalts (OIB) from Hawaii, Samoa, and Iceland reveal variable 182W/184W, ranging from that of the ambient upper mantle to ratios as much as 18 parts per million lower. The tungsten isotopic data negatively correlate with 3He/4He. These data indicate that each OIB system accesses domains within Earth that formed within the first 60 million years of solar system history. Combined isotopic and chemical characteristics projected for these ancient domains indicate that they contain metal and are repositories of noble gases. We suggest that the most likely source candidates are mega-ultralow-velocity zones, which lie beneath Hawaii, Samoa, and Iceland but not beneath hot spots whose OIB yield normal 182W and homogeneously low 3He/4He.


Isofagomine in vivo effects in a neuronopathic Gaucher disease mouse.
The pharmacological chaperone, isofagomine (IFG), enhances acid β-glucosidase (GCase) function by altering folding, trafficking, and activity in wild-type and Gaucher disease fibroblasts. The in vivo effects of IFG on GCase activity, its substrate levels, and phenotype were evaluated using a neuronopathic Gaucher disease mouse model, 4L;C* (V394L/V394L + saposin C-/-) that has CNS accumulation of glucosylceramide (GC) and glucosylsphingosine (GS) as well as progressive neurological deterioration. IFG administration to 4L;C* mice at 20 or 600 mg/kg/day resulted in life span extensions of 10 or 20 days, respectively, and increases in GCase activity and protein levels in the brain and visceral tissues. Cerebral cortical GC and GS levels showed no significant reductions with IFG treatment. Increases of GC or GS levels were detected in the visceral tissues of IFG treated (600 mg/kg/day) mice. The attenuations of brain proinflammatory responses in the treated mice were evidenced by reductions in astrogliosis and microglial cell activation, and decreased p38 phosphorylation and TNFα levels. Terminally, axonal degeneration was present in the brain and spinal cord from untreated and treated 4L;C* mice. These data demonstrate that IFG exerts in vivo effects by enhancing V394L GCase protein and activity levels, and in mediating suppression of proinflammation, which led to delayed onset of neurological disease and extension of the life span of 4L;C* mice. However, this was not correlated with a reduction in the accumulation of lipid substrates.


The lectin-like domain of tumor necrosis factor-alpha increases membrane conductance in microvascular endothelial cells and peritoneal macrophages.
Herein, we show that TNF exerts a pH-dependent increase in membrane conductance in primary lung microvascular endothelial cells and peritoneal macrophages. This effect was TNF receptor-independent, since it also occurred in cells isolated from mice deficient in both types of TNF receptors. A TNF mutant in which the three amino acids critical for the lectin-like activity were replaced by an alanine did not show any significant effect on membrane conductance. Moreover, a synthetic 17-amino acid peptide of TNF, which was previously shown to exert lectin-like activity, also increased the ion permeability in these cells. The amiloride sensitivity of the observed activity suggests a binding of TNF to an endogenous ion channel rather than channel formation by TNF itself. This may have important implications in mechanisms of TNF-mediated vascular pathology.


Portal venous system thrombosis: helical CT angiography before transjugular intrahepatic portosystemic shunt creation.
To evaluate the utility of helical computed tomographic (CT) angiography for depiction of thrombi in the portal venous system in patients under consideration for transjugular intrahepatic portosystemic shunt (TIPS) creation. Contrast material-enhanced helical CT was performed before TIPS creation in 25 patients. Axial, multiplanar, and three-dimensional images were evaluated to determine whether thrombus was present in the portal system and whether TIPS creation was contraindicated. CT findings were confirmed at visceral angiography (n = 3), direct portography (n = 20), or duplex ultrasonography (n = 2). Ten (40%) of 25 patients, including 10 (56%) of 18 patients with refractory variceal hemorrhage, had thrombus in the portal venous system. Helical CT scans depicted thrombus in nine (90%) of 10 patients (95% confidence interval = 0.71, 1.00) and in 16 (94%) of 17 vessels (95% confidence interval = 0.83, 1.00), including the portal vein (eight of eight patients), splenic vein (three of four patients), and superior mesenteric vein (five of five patients). TIPS creation was canceled in four (16%) patients on the basis of CT findings. Thrombi in the portal venous system are common in patients with refractory variceal hemorrhage. Helical CT angiography is sensitive and specific for portal venous system thrombosis and can provide information that alters treatment in these patients.


Patient Interest in Receiving Assistance with Self-Reported Social Risks.
This study evaluated how often patients who reported social risk factors requested assistance with these risks in an integrated health system. We examined how self-reports of risk related to stated desire for help with that risk reported during social risk screenings at Kaiser Permanente Northwest (KPNW). We examined how patient characteristics were associated with desire for help with each social risk domain using logistic regression. Approximately 24% (n = 7,807) of the 32,865 KPNW members aged ≥ 18 years who were screened between June 1, 2017, and December 31, 2019, reported at least 1 social risk. More than half of patients who reported a risk were risk/help concordant (i.e., they also wanted help with that risk). The highest concordance (81.7%) was observed among patients reporting medical financial hardship. Several demographic, health, and other factors were associated with concordance across domains. Patients do not request assistance for all reported social needs. Our findings could help shape future work examining patients' reasons for not accepting assistance and developing interventions to help patients with high social risk more effectively.


Market forces and efficient health care systems.
The "market forces" to which economists ascribe the ability to motivate improvement in quality and efficiency are largely nonexistent in U.S. health care. One thus might ask, "Could market forces be made strong enough to deliver efficient health care systems?" There is some evidence to suggest that the answer is "Yes." This paper offers a short list of some changes that would be needed to create such a health care economy. Continued increases in costs and in the numbers of uninsured people will likely make a universal coverage model based on Medicare a politically popular choice, but such a model would not deliver efficient health care systems because it lacks sufficient incentives for consumers to choose less costly options.


THE ROLE OF DISULFIDE BONDS IN THE COMPLEMENT-FIXING AND PRECIPITATING PROPERTIES OF 7S RABBIT AND SHEEP ANTIBODIES.
The number of total disulfide bonds in rabbit and sheep 7S gamma globulin, before and after treatment with 2-mercaptoethanol, has been measured by amperometric titration. Mercaptan reduction could diminish the complement-fixing efficiency of 7S rabbit gamma globulin by no more than 90 per cent without any significant decrease in maximal complement-fixing ability. This was associated with the reduction of only 7 disulfide bonds, including 1 interchain disulfide bond [S-S((1))]. The reduction of 7 disulfide bonds in 7S sheep gamma globulin was associated with a 64 per cent decrease in complement-fixing efficiency, while reduction of 10 to 11 disulfide bonds decreased the complement-fixing efficiency by 90 per cent. Reduction of more disulfide bonds was not associated with any further decrease in complement-fixing efficiency, nor of any decrease in maximal complement-fixing ability. The reduction of more than 10 to 11 disulfide bonds in 7S rabbit and sheep gamma globulins was associated with a decrease in precipitating ability. The disulfide bonds which are more labile to mercaptan reduction appear to be associated with complement-fixing efficiency while the disulfide bonds which are more resistant to mercaptan reduction appear to be associated with precipitating ability. The measurement of interpolypeptide chain disulfide bonds has been discussed. One easily reduced interchain disulfide bond [S-S((1))] appears to be important for the complement-fixing efficiency of 7S rabbit antibody. The integrity of this same bond is essential for the precipitating ability of 5S rabbit antibody and may also be important for its complement-fixing ability.


Infants and Gin Mania in 18th-century London.
A study of Hogarth's engraving, Gin Lane, highlights the fact that alcoholic addiction parents results not only in the fetal alcohol syndrome, but also in other types of infant abuse. Physical trauma, loss of parents, neglect, and malnutrition are all depicted. There is the strong implication that the causes of such infant abuse, and thus the prevention, lie largely in socioeconomic and cultural fields, rather than in that of medical science.


Fabrication of 3D high aspect ratio PDMS microfluidic networks with a hybrid stamp.
We report a novel methodology for fabricating large-area, multilayer, thin-film, high aspect ratio, 3D microfluidic structures with through-layer vias and open channels that can be bonded between hard substrates. It is realized by utilizing a hybrid stamp with a thin plastic sheet embedded underneath a PDMS surface. This hybrid stamp solves an important edge protrusion issue during PDMS molding while maintaining necessary stamp elasticity to ensure the removal of PDMS residues at through-layer regions. Removing edge protrusion is a significant progress toward fabricating 3D structures since high aspect ratio PDMS structures with flat interfaces can be realized to facilitate multilayer stacking and bonding to hard substrates. Our method also allows for the fabrication of 3D deformable channels, which can lead to profound applications in electrokinetics, optofluidics, inertial microfluidics, and other fields where the shape of the channel cross section plays a key role in device physics. To demonstrate, as an example, we have fabricated a microfluidic channel by sandwiching two 20 μm wide, 80 μm tall PDMS membranes between two featureless ITO glass substrates. By applying electrical bias to the two ITO substrates and pressure to deform the thin membrane sidewalls, strong electric field enhancement can be generated in the center of a channel to enable 3D sheathless dielectrophoretic focusing of biological objects including mammalian cells and bacteria at a flow speed up to 14 cm s(-1).


Secondary pulmonary hypertension in chronic heart failure: the role of the endothelium in pathophysiology and management.
Pulmonary vascular resistance is frequently elevated in patients with chronic left ventricular failure as a result of dysregulation of vascular smooth muscle tone and structural remodeling. The former is reversible over a period of minutes to days by pharmacological vasodilators, whereas the latter is relatively fixed and may resolve only slowly, over months to years. These abnormalities are due, at least in part, to pulmonary vascular endothelial dysfunction that results in impaired nitric oxide availability and increased endothelin expression. In patients with chronic heart failure, the resulting pulmonary hypertension directly affects right ventricular function and may affect exercise capacity, morbidity, and mortality. New treatment strategies, which include the use of agents that increase nitric oxide availability or oppose the actions of endothelin, may improve the structure and function of the pulmonary vasculature in patients with chronic heart failure.


A Bayesian approach for characterizing direction tuning curves in the supplementary motor area of behaving monkeys.
Neural responses are commonly studied in terms of "tuning curves," characterizing changes in neuronal response as a function of a continuous stimulus parameter. In the motor system, neural responses to movement direction often follow a bell-shaped tuning curve for which the exact shape determines the properties of neuronal movement coding. Estimating the shape of that tuning curve robustly is hard, especially when directions are sampled unevenly and at a coarse resolution. Here, we describe a Bayesian estimation procedure that improves the accuracy of curve-shape estimation even when the curve is sampled unevenly and at a very coarse resolution. Using this approach, we characterize the movement direction tuning curves in the supplementary motor area (SMA) of behaving monkeys. We compare the SMA tuning curves to tuning curves of neurons from the primary motor cortex (M1) of the same monkeys, showing that the tuning curves of the SMA neurons tend to be narrower and shallower. We also show that these characteristics do not depend on the specific location in each region.