Return of Research-Related Genetic Test Results and Genetic Discrimination Concerns: Facilitators and Barriers of Genetic Research Participation in Diverse Groups.
Most genetics studies lack the diversity necessary to ensure that all groups benefit from genetic research. To explore facilitators and barriers to genetic research participation. We conducted a survey on genetics in research and healthcare from November 15, 2017 to February 28, 2018 among adult Kaiser Permanente (KP) members who had been invited to participate in the KP biobank (KP Research Bank). We used logistic regression to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CIs) comparing the willingness to participate in genetic research under different return of results scenarios and genetic discrimination concerns between groups, according to their demographic characteristics. A total of 57,331 KP members were invited to participate, and 10,369 completed the survey (18% response rate). Respondents were 65% female, 44% non-Hispanic White (NH White), 22% Asian/Native Hawaiian or other Pacific Islander (Asian/PI), 19% non-Hispanic Black (NH Black), and 16% Hispanic. Respondents willing to participate in genetic research ranged from 22% with no results returned to 87% if health-related genetic results were returned. We also found variation by race/ethnicity; when no results were to be returned, Asian/PIs, Hispanics, and NH Blacks were less likely to want to participate than NH Whites (p < 0.05). However, when results were returned, disparities in the willingness to participate disappeared for NH Blacks and Hispanics. Genetic discrimination concerns were more prevalent in Asian/PIs, Hispanics, and NH Blacks than in NH Whites (p < 0.05). Policies that prohibit the return of results and do not address genetic discrimination concerns may contribute to a greater underrepresentation of diverse groups in genetic research.


Near Infrared Light Treatment Reduces Synaptic Levels of Toxic Tau Oligomers in Two Transgenic Mouse Models of Human Tauopathies.
Tau oligomers are emerging as a key contributor to the synaptic dysfunction that drives cognitive decline associated with the clinical manifestation and progression of Alzheimer's disease (AD). Accordingly, there is ample consensus that interventions that target tau oligomers may slow or halt the progression of AD. With this ultimate goal in mind, in the present study, we investigated tau oligomer accumulation and its synaptic and behavioral consequences after an in vivo treatment with near infrared (NIR) light (600-1000 nm) in two transgenic mouse models, overexpressing human tau either alone (hTau mice) or in combination with amyloid beta (3xTgAD mice). We found that a 4-week exposure to NIR light (90 s/day/5 days a week) significantly reduced levels of endogenous total and oligomeric tau in both synaptosomes and total protein extracts from the hippocampus and cortex of hTau mice and improved deteriorating memory function. Similar results were observed in the 3xTgAD mice, which further displayed reduced synaptic Aβ after NIR light treatment. On the other hand, ex vivo binding of tau oligomers in isolated synaptosomes as well as tau oligomer-induced depression of long-term potentiation (LTP) in hippocampal slices from NIR light-treated wt mice were unaffected. Finally, levels of proteins critically involved in two mechanisms associated with clearance of misfolded tau, inducible HSP70 and autophagy, were upregulated in NIR light treated mice. Collectively, these results show that NIR light decreases levels of endogenous toxic tau oligomers and alleviate associated memory deficits, thus furthering the development of NIR light as a possible therapeutic for AD.


Remembering where we're from: Community- and individual-level predictors of college students' White privilege awareness.
Scholars in the field of community psychology have called for more research dedicated to examining White privilege as part of a system of White supremacy in the United States. One branch of this work focuses on awareness of White privilege, yet to date, this research has typically investigated awareness of White privilege at individual levels of analysis instead of also focusing on neighborhoods, schools, and other levels of analysis beyond the individual. In this study, we combine survey and U.S. Census data to explore both individual- and community-level predictors of White privilege awareness. With a sample of 1285 White college students, we found that gender, modern racism, social dominance orientation, and subjective socioeconomic status (SES) significantly predicted White privilege awareness. After accounting for these individual-level variables, we found that characteristics of students' hometowns (defined by zip code) predicted White privilege awareness. Specifically, greater income inequality was associated with higher White privilege awareness, while greater White racial homogeneity was marginally associated with lower White privilege awareness. There was a significant interaction between community-level White racial homogeneity and individual-level subjective SES, such that students with high subjective SES and low White racial homogeneity had the highest White privilege awareness. This study highlights the importance of examining different facets of ecological context in relation to White Americans' racial attitudes.


Still Searching for a Suitable Molecular Test to Detect Hidden Plasmodium Infection: A Proposal for Blood Donor Screening in Brazil.
Efforts have been made to establish sensitive diagnostic tools for malaria screening in blood banks in order to detect malaria asymptomatic carriers. Microscopy, the malaria reference test in Brazil, is time consuming and its sensitivity depends on microscopist experience. Although molecular tools are available, some aspects need to be considered for large-scale screening: accuracy and robustness for detecting low parasitemia, affordability for application to large number of samples and flexibility to perform on individual or pooled samples. In this retrospective study, we evaluated four molecular assays for detection of malaria parasites in a set of 56 samples previously evaluated by expert microscopy. In addition, we evaluated the effect of pooling samples on the sensitivity and specificity of the molecular assays. A well-characterized cultured sample with 1 parasite/μL was included in all the tests evaluated. DNA was extracted with QIAamp DNA Blood Mini Kit and eluted in 50 μL to concentrate the DNA. Pools were assembled with 10 samples each. Molecular protocols targeting 18S rRNA, included one qPCR genus specific (Lima-genus), one duplex qPCR genus/Pf (PET-genus, PET-Pf) and one duplex qPCR specie-specific (Rougemont: Roug-Pf/Pv and Roug-Pm/Po). Additionally a nested PCR protocol specie-specific was used (Snou-Pf, Snou-Pv, Snou-Pm and Snou-Po). The limit of detection was 3.5 p/μL and 0.35p/μl for the PET-genus and Lima-genus assays, respectively. Considering the positive (n = 13) and negative (n = 39) unpooled individual samples according to microscopy, the sensitivity of the two genus qPCR assays was 76.9% (Lima-genus) and 72.7% (PET-genus). The Lima-genus and PET-genus showed both sensitivity of 86.7% in the pooled samples. The genus protocols yielded similar results (Kappa value of 1.000) in both individual and pooled samples. Efforts should be made to improve performance of molecular tests to enable the detection of low-density parasitemia if these tests are to be utilized for blood transfusion screening.


Inhibition of mitochondrial carrier homolog 2 (MTCH2) suppresses tumor invasion and enhances sensitivity to temozolomide in malignant glioma.
Malignant glioma exerts a metabolic shift from oxidative phosphorylation (OXPHOs) to aerobic glycolysis, with suppressed mitochondrial functions. This phenomenon offers a proliferation advantage to tumor cells and decrease mitochondria-dependent cell death. However, the underlying mechanism for mitochondrial dysfunction in glioma is not well elucidated. MTCH2 is a mitochondrial outer membrane protein that regulates mitochondrial metabolism and related cell death. This study aims to clarify the role of MTCH2 in glioma. Bioinformatic analysis from TCGA and CGGA databases were used to investigate the association of MTCH2 with glioma malignancy and clinical significance. The expression of MTCH2 was verified from clinical specimens using real-time PCR and western blots in our cohorts. siRNA-mediated MTCH2 knockdown were used to assess the biological functions of MTCH2 in glioma progression, including cell invasion and temozolomide-induced cell death. Biochemical investigations of mitochondrial and cellular signaling alternations were performed to detect the mechanism by which MTCH2 regulates glioma malignancy. Bioinformatic data from public database and our cohort showed that MTCH2 expression was closely associated with glioma malignancy and poor patient survival. Silencing of MTCH2 expression impaired cell migration/invasion and enhanced temozolomide sensitivity of human glioma cells. Mechanistically, MTCH2 knockdown may increase mitochondrial OXPHOs and thus oxidative damage, decreased migration/invasion pathways, and repressed pro-survival AKT signaling. Our work establishes the relationship between MTCH2 expression and glioma malignancy, and provides a potential target for future interventions.


Structure of aryl O-demethylase offers molecular insight into a catalytic tyrosine-dependent mechanism.
Some strains of soil and marine bacteria have evolved intricate metabolic pathways for using environmentally derived aromatics as a carbon source. Many of these metabolic pathways go through intermediates such as vanillate, 3-O-methylgallate, and syringate. Demethylation of these compounds is essential for downstream aryl modification, ring opening, and subsequent assimilation of these compounds into the tricarboxylic acid (TCA) cycle, and, correspondingly, there are a variety of associated aryl demethylase systems that vary in complexity. Intriguingly, only a basic understanding of the least complex system, the tetrahydrofolate-dependent aryl demethylase LigM from Sphingomonas paucimobilis, a bacterial strain that metabolizes lignin-derived aromatics, was previously available. LigM-catalyzed demethylation enables further modification and ring opening of the single-ring aromatics vanillate and 3-O-methylgallate, which are common byproducts of biofuel production. Here, we characterize aryl O-demethylation by LigM and report its 1.81-Å crystal structure, revealing a unique demethylase fold and a canonical folate-binding domain. Structural homology and geometry optimization calculations enabled the identification of LigM's tetrahydrofolate-binding site and protein-folate interactions. Computationally guided mutagenesis and kinetic analyses allowed the identification of the enzyme's aryl-binding site location and determination of its unique, catalytic tyrosine-dependent reaction mechanism. This work defines LigM as a distinct demethylase, both structurally and functionally, and provides insight into demethylation and its reaction requirements. These results afford the mechanistic details required for efficient utilization of LigM as a tool for aryl O-demethylation and as a component of synthetic biology efforts to valorize previously underused aromatic compounds.


Control of human beta-globin mRNA stability and its impact on beta-thalassemia phenotype.
Messenger RNA (mRNA) stability is a critical determinant that affects gene expression. Many pathways have evolved to modulate mRNA stability in response to developmental, physiological and/or environmental stimuli. Eukaryotic mRNAs have a considerable range of half-lives, from as short as a few minutes to as long as several days. Human globin mRNAs constitute an example of highly stable mRNAs. However, a wide variety of naturally occurring mutations that result in the clinical syndrome of thalassemia can trigger accelerated mRNA decay thus controlling mRNA quality prior to translation. Distinct surveillance mechanisms have been described as being targeted for specific defective globin mRNAs. Here, we review mRNA stability mechanisms implicated in the control of β-globin gene expression and the surveillance pathways that prevent translation of aberrant β-globin mRNAs. In addition, we emphasize the importance of these pathways in modulating the severity of the β-thalassemia phenotype.


Causal effects of life course adiposity on chronic kidney disease: a Mendelian randomization study.
Obesity is reported as closely correlated with the development of chronic kidney disease (CKD); however, whether causation exists is unknown, and controversy remains about whether the role of obesity is protective or destructive in CKD. In this study, we attempted to infer the causal relationship between life course adiposity and CKD, to provide a rationale for obesity management in CKD patients. A 2-sample Mendelian randomization (MR) analysis was conducted to explore the causal relationship of life course adiposity traits including body mass index (BMI), childhood BMI, body fat percentage (BF%), birth weight (BW), waist circumference, hip circumference, and waist-to-hip ratio (WHR) to CKD. Significant single nucleotide polymorphisms from genome-wide association study on human adiposity traits were utilized as exposure instruments, and summary statistics of CKD as the outcome. The causal relationship was evaluated by inverse variance weighted, MR Egger regression and weighted median methods, and further verified by extensive sensitivity analyses. Genetically determined one standard deviation increase in adult BMI was associated with higher risk of CKD in all 4 MR methods. Other indexes including childhood BMI, body fat percentage, and waist/hip circumference were also shown to have a causal effect on the risk of CKD. The results were robust under all sensitivity analyses. There exists a causal effect of life course adiposity on the risk of CKD. A genetic predisposition to higher adult BMI may increase the risk of CKD.


The calcium-binding protein parvalbumin modulates the firing 1 properties of the reticular thalamic nucleus bursting neurons.
The reticular thalamic nucleus (RTN) of the mouse is characterized by an overwhelming majority of GABAergic neurons receiving afferences from both the thalamus and the cerebral cortex and sending projections mainly on thalamocortical neurons. The RTN neurons express high levels of the "slow Ca(2+) buffer" parvalbumin (PV) and are characterized by low-threshold Ca(2+) currents, I(T). We performed extracellular recordings in ketamine/xylazine anesthetized mice in the rostromedial portion of the RTN. In the RTN of wild-type and PV knockout (PVKO) mice we distinguished four types of neurons characterized on the basis of their firing pattern: irregular firing (type I), medium bursting (type II), long bursting (type III), and tonically firing (type IV). Compared with wild-type mice, we observed in the PVKOs the medium bursting (type II) more frequently than the long bursting type and longer interspike intervals within the burst without affecting the number of spikes. This suggests that PV may affect the firing properties of RTN neurons via a mechanism associated with the kinetics of burst discharges. Ca(v)3.2 channels, which mediate the I(T) currents, were more localized to the somatic plasma membrane of RTN neurons in PVKO mice, whereas Ca(v)3.3 expression was similar in both genotypes. The immunoelectron microscopy analysis showed that Ca(v)3.2 channels were localized at active axosomatic synapses, thus suggesting that the differential localization of Ca(v)3.2 in the PVKOs may affect bursting dynamics. Cross-correlation analysis of simultaneously recorded neurons from the same electrode tip showed that about one-third of the cell pairs tended to fire synchronously in both genotypes, independent of PV expression. In summary, PV deficiency does not affect the functional connectivity between RTN neurons but affects the distribution of Ca(v)3.2 channels and the dynamics of burst discharges of RTN cells, which in turn regulate the activity in the thalamocortical circuit.


Medication Administration Errors in a University Hospital.
To measure the rates of medication administration errors in medical wards at Ain Shams University Hospital and to identify significant determinants of medication administration errors. A descriptive direct-observational study of drug administration errors was carried out at medical wards of Ain Shams University hospital for a period of 3 months. A standardized observational checklist was used to observe the nurse during giving medications, and a medical record audit form was used to assess documentation. The error rates per observation, nurse, and patient were calculated, and the association between error rates and characteristics of each category was tested using linear regression to identify potential risk factors. The study included 237 patients and 28 nurses. The final number of drug administration observations was 2090 after excluding 310 omissions. A total of 5531 errors were observed with an average number of 2.67 errors per observation. More than 85% of the observations had at least one error, and the overall error rate was 37.68% (per hundred error opportunities). The highest error rate was detected in injections especially the intravenous route (39.58%). The most frequent errors were wrong documentation (90.96%) and wrong technique (78.90%), and the least was wrong patient (0.05%). The significant independent determinants of medication administration errors were high number of shifts taken by nurse per month, night shifts, weekends, elderly patient, and illiteracy. Medication administration errors represent a major problem in the hospital that needs urgent intervention to optimize medication administration process. The intervention should consider the identified significant determinants of medication administration errors.


Effect of postpartum collection time and colostrum quality on passive transfer of immunity, performance, and small intestinal development in preweaning calves.
This study evaluated the effects of postpartum collection time and quality of colostrum fed to calves on the failure of passive transfer, growth, and small intestine development in the first 5 wk of life. Newborn calves (Holstein-Friesian × Jersey) were identified at birth and collected either early (E; within 12 h postpartum; n = 20) or late (L; 18-24 h postpartum; n = 20) and fed either high-quality colostrum [HQC, first milking colostrum with Brix% = 23 ± standard deviation (SD) 2] or low-quality colostrum (LQC, mixed colostrum and transition milk with Brix% = 12 ± 1) to create 4 treatments: E-HQC, E-LQC, L-HQC, and L-LQC (n = 10/treatment). After collection, calves (body weight = 32.3 ± 4.6 kg/calf) were fed either HQC or LQC (7.5% of their arrival body weight per feed) for the first 3 (L calves) or 4 feedings (E calves). All calves were then managed and fed similarly using automatic feeders which recorded individual intake of milk replacer and calf starter. Blood samples were taken at d 1 (after collection from dams but before colostrum feeding), 4, 14, and 35 of age to analyze selected metabolites. All calves were killed at d 35 ± 2 of age and histomorphology of duodenum, jejunum, and ileum was evaluated. At collection, 75% of E calves and 58% of L calves had serum total protein ≤52 g/L. At d 4 of age, calves fed HQC had greater serum total protein than calves fed LQC; however, failure of passive transfer (serum total protein ≤52 g/L) incidence did not differ between HQC and LQC. Collection time did not affect the scouring duration, but the amount of electrolyte used to treat sick calves was lower in L versus E calves, whereas feeding HQC versus LQC lowered both the scouring duration and electrolyte use to treat sick calves. Calves fed HQC had a greater total surface area of the duodenum (+23%) and jejunum (+17%) compared with LQC calves. Duodenal crypts were deeper in E-LQC calves than E-HQC and L-HQC calves, whereas L-LQC calves were intermediate. Villus height to crypt depth ratio in duodenum, jejunum, and ileum was greater in HQC than LQC calves. A trend toward greater average daily gain was observed in HQC versus LQC calves (667 vs. 590 g/d) but the average daily gain was not influenced by collection time. Serum IGF-1 at d 4 was higher in HQC versus LQC calves and this might have contributed to greater average daily gain and small intestine development. Calves fed HQC had higher feed conversion ratios (FCR; total body weight gain/total dry matter intake) compared with LQC calves, and L calves had higher FCR compared with E calves. In conclusion, in comparison to feeding LQC, feeding HQC reduced the scouring duration, enhanced surface area of duodenum and jejunum, and improved FCR during the first 5 wk of calf age. Postpartum collection time of calves did not affect small intestine development, but L calves had higher FCR and required a lesser volume of electrolytes to treat scours compared with E calves during the first 35 d of life.


Suppression of stimulated plasma renin by clonidine in the dog.
The mechanism by which clonidine suppresses plasma renin activity (PRA) was investigated in dogs anesthetized with pentobarbital. Injection of clonidine (1 micrograms/kg) into the cisterna magna decreased PRA from levels stimulated by prior hemorrhage into a blood reservoir to reduce mean blood pressure by 25% (21.7 ng/ml/hr +/- 6.6 SE leads to 11.1 ng/ml/hr +/- 2.4 SE; p less than 0.05). Clonidine reduced heart rate but mean arterial pressure remained constant due to fluid movement between the reservoir and the arterial circulation of the dog. These effects could not be attributed to leakage of clonidine from the cerebrospinal fluid since intravenous administration of the same dose had no effect on PRA. In animals bilaterally splanchnicotomized at the level of the diaphragm, elevated PRA was not reduced by intracisternal clonidine. When return of reservoir fluid was prevented, animals became hypotensive after central clonidine and renin tended to increase. These results indicate that clonidine reduces stimulated renin by a central mechanism that is dependent upon the integrity of the sympathetic innervation of the kidney. Other stimuli for renin release may override the inhibitory effect of central clonidine.


[Nutritional characteristics of school lunch menus in Biscay (Basque Country, Spain) in 2012/2013].
One of the areas of intervention of the Spanish strategy for nutrition, physical activity and prevention of obesity (NAOS) is school nutrition. To evaluate, taking as reference the minimum set of indicators and recommendations of NAOS Strategy, a sample of school lunch menus distributed in several school dining rooms of Biscay. A descriptive cross sectional study was conducted, based on the theoretical information of the basal menus distributed by seven catering enterprises to 324 schools (80,969 pupils). Weekly frequency of several types of food groups was determined and evaluated; reference documents were "Evaluation and Monitoring of NAOS Strategy: set of indicators" and recommendations of the "Consensus document of feeding in school". The 100 per cent of the school dining rooms meets minimums concerning legumes, fish and pre-cooked meals and with the recommendations of meat and meat products. However, although being close to guidelines, none meets the minimum of fruit, vegetables or of additional nutritional information. Differences among NAOS Strategy recommendations and requirements of the Department of Education have been found, and also difficulties in the quantification and classification of several foodstuffs. It is recommended to increase the content of fruits, vegetables, eggs and oily fish in school menus as well as offering more detailed description of menus to families.


Determination of porphobilinogen deaminase activity in human erythrocytes: pertinent factors in obtaining optimal conditions for measurements.
Determination of porphobilinogen deaminase (PBGD; EC 184.108.40.206) activity in erythrocytes can contribute to the identification of patients suspected of acute intermittent porphyria. PBGD catalyses the polymerization of four molecules of porphobilinogen (PBG) to the highly unstable 1-hydroxymethylbilane. The 1-hydroxymethylbilane is transformed into uroporphyrinogen III by uroporphyrinogen III synthase. When this enzyme is inactivated, 1-hydroxymethylbilane cyclizes non-enzymatically to uroporphyrinogen I, which can be oxidized to uroporphyrin I. PBGD activity can be measured by quantitation of uroporphyrin I formed from PBG under conditions where this is the only end product. The purpose of the present study was to define the optimal conditions for quantitating PBGD activity in human erythrocytes. The preanalytical factors examined were: anticoagulants and methods for disruption of the erythrocytes. The analytical factors examined were: duration of preincubation, reaction time, reaction temperature, pH, ionic strength and conditions for the oxidation of uroporphyrinogen I to uroporphyrin I. Based on the results, we propose an optimized method for determination of PBGD activity in erythrocytes.


Fusion of Ferredoxin and Cytochrome P450 Enables Direct Light-Driven Biosynthesis.
Cytochrome P450s (P450s) are key enzymes in the synthesis of bioactive natural products in plants. Efforts to harness these enzymes for in vitro and whole-cell production of natural products have been hampered by difficulties in expressing them heterologously in their active form, and their requirement for NADPH as a source of reducing power. We recently demonstrated targeting and insertion of plant P450s into the photosynthetic membrane and photosynthesis-driven, NADPH-independent P450 catalytic activity mediated by the electron carrier protein ferredoxin. Here, we report the fusion of ferredoxin with P450 CYP79A1 from the model plant Sorghum bicolor, which catalyzes the initial step in the pathway leading to biosynthesis of the cyanogenic glucoside dhurrin. Fusion with ferredoxin allows CYP79A1 to obtain electrons for catalysis by interacting directly with photosystem I. Furthermore, electrons captured by the fused ferredoxin moiety are directed more effectively toward P450 catalytic activity, making the fusion better able to compete with endogenous electron sinks coupled to metabolic pathways. The P450-ferredoxin fusion enzyme obtains reducing power solely from its fused ferredoxin and outperforms unfused CYP79A1 in vivo. This demonstrates greatly enhanced electron transfer from photosystem I to CYP79A1 as a consequence of the fusion. The fusion strategy reported here therefore forms the basis for enhanced partitioning of photosynthetic reducing power toward P450-dependent biosynthesis of important natural products.


Curative one-shot systemic virotherapy in murine myeloma.
Current therapy for multiple myeloma is complex and prolonged. Antimyeloma drugs are combined in induction, consolidation and/or maintenance protocols to destroy bulky disease, then suppress or eradicate residual disease. Oncolytic viruses have the potential to mediate both tumor debulking and residual disease elimination, but this curative paradigm remains unproven. Here, we engineered an oncolytic vesicular stomatitis virus to minimize its neurotoxicity, enhance induction of antimyeloma immunity and facilitate noninvasive monitoring of its intratumoral spread. Using high-resolution imaging, autoradiography and immunohistochemistry, we demonstrate that the intravenously administered virus extravasates from tumor blood vessels in immunocompetent myeloma-bearing mice, nucleating multiple intratumoral infectious centers that expand rapidly and necrose at their centers, ultimately coalescing to cause extensive tumor destruction. This oncolytic tumor debulking phase lasts only for 72 h after virus administration, and is completed before antiviral antibodies become detectable in the bloodstream. Antimyeloma T cells, cross-primed as the virus-infected cells provoke an antiviral immune response, then eliminate residual uninfected myeloma cells. The study establishes a curative oncolytic paradigm for multiple myeloma where direct tumor debulking and immune eradication of minimal disease are mediated by a single intravenous dose of a single therapeutic agent. Clinical translation is underway.


Down-regulation of promoter 1.3 activity of the human aromatase gene in breast tissue by zinc-finger protein, snail (SnaH).
Aromatase (estrogen synthetase) is expressed in breast cancer tissue, and in situ expression of the enzyme stimulates breast cancer growth. Promoter I.3 is one of the major promoters that control the expression of aromatase in breast cancer tissue. Using the yeast one-hybrid approach to screen a human breast tissue hybrid cDNA expression library, we found that the zinc-finger transcriptional factor Snail (SnaH) interacted with a regulatory region near promoter I.3 of the human aromatase gene. DNA mobility shift assays and mutation analyses using recombinant SnaH protein expressed in Escherichia coli have revealed that this protein interacts with a segment, 5'-CTGATGAAGT-3', which is between 66 and 76 bp upstream from the transcriptional start site of promoter I.3. Using mammalian cell transfection experiments, SnaH was found to act as a repressor of promoter I.3 activity. Site-directed mutagenesis experiments have revealed that the NH2-terminal SNAG domain is important for the repressor activity of SnaH. To demonstrate the inhibitory activity against aromatase expression, a stable SnaH-expressing MDA-MB-231 breast cancer cell line was generated, and the aromatase RNA messages in the SnaH-transfected cell line were found to be 30% of those in the vector-transfected cell line. Reverse transcription-PCR analysis on RNAs isolated from 12 cell lines has confirmed that SnaH is expressed at a higher level in normal breast epithelial cell and stromal fibroblast cell lines than in breast cancer cell lines. In addition, SnaH mRNA was detected in only 16 of 55 breast cancer specimens. On the other hand, aromatase mRNA was detected in 54 of the 55 specimens. Our results indicate that SnaH acts as a repressor that down-regulates the expression of aromatase in normal breast tissue by suppressing the function of promoter I.3. A reduction of the expression of SnaH in breast cancer tissue further suggests a cancer-protective role for this protein in normal breast tissue.


[The enterosorbents belosorb and panzysorb in the treatment of acute experimental pancreatitis in rats].
Enterosorption with highly dispersed fibrous carbon adsorbent Belosorb or enzyme-containing fibrous carbon adsorbent Panzysorb was applied for the treatment of experimental acute pancreatitis (EAP) in the rats. Panzysorb was obtained by immobilization of digestive hydrolases (proteases and amylase) on Belosorb as matrix. It was shown that oral administration of Panzysorb (100 mg/kg body weight) significantly decreased amylolytic and lipolytic activity of blood serum by the third hour after EAP induction. The course of enterosorption (ten days) with the Panzysorb led to the appearance of regeneration focuses in injured organ.


Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment.
DILI is a major safety issue during drug development and one of the leading causes for market withdrawal. Despite many efforts made in the past, the prediction of DILI using in vitro models remains very unreliable. In the present study, the well-established hepatocyte Collagen I-Matrigel™ sandwich culture was used, mimicking chronic drug treatment after multiple incubations for 14 days. Ten drugs associated with different types of specific preclinical and clinical liver injury were evaluated at non-cytotoxic concentrations. Mrp2-mediated transport, intracellular accumulation of neutral lipids and phospholipids were selected as functional endpoints by using Cellomics™ Arrayscan® technology and assessed at five timepoints (day 1, 3, 7, 10, 14). Liver specific functional impairments after drug treatment were enhanced over time and could be monitored by HCI already after few days and before cytotoxicity. Phospholipidosis-inducing drugs Chlorpromazine and Amiodarone displayed the same response as in vivo. Cyclosporin A, Chlorpromazine, and Troglitazone inhibited Mrp2-mediated biliary transport, correlating with in vivo findings. Steatosis remained difficult to be reproduced under the current in vitro testing conditions, resulting into false negative and positive responses. The present results suggest that the repeated long-term treatment of rat hepatocytes in the Collagen I-Matrigel™ sandwich configuration might be a suitable tool for safety profiling of the potential to induce phospholipidosis and impair Mrp2-mediated transport processes, but not to predict steatosis.


Exploring cardiac arrest in 'at-home' settings: concepts derived from a qualitative interview study with layperson bystanders.
Two thirds of Out-of-Hospital Cardiac Arrests (OHCAs) occur at the patient's home ('at-home-CA'), where bystander CPR (B-CPR) rates are significantly lower than in public locations. Knowledge about the circumstances of this specific setting has mainly been limited to quantitative data. To develop a more conceptual understanding of the circumstances and dynamics of 'at-home CA', we conducted a qualitative interview study. Twenty-one semi-structured in-depth interviews were performed with laypersons who had witnessed 'at-home CA'. The interviews were audio recorded, transcribed, and analysed by qualitative content analysis (QCA). A category system was developed to classify facilitating and impeding factors and to finally derive overarching concepts of 'at-home CA'. Qualitative Content Analysis yielded 1'347 relevant interview segments. Of these, 398 related to factors facilitating B-CPR, 328 to factors impeding, and 621 were classified neutral. Some of these factors were specific to 'at-home CA'. The privacy context was found to be a particularly supportive factor, as it enhanced the commitment to act and facilitated the detection of symptoms. Impeding factors, aggravated in 'at-home CA' settings, included limited support from other bystanders, acute stress response and impaired situational judgement, as well as physical challenges when positioning the patient. We derived six overarching concepts defining the 'at-home CA' situation: (a) unexpectedness of the event, (b) acute stress response, (c) situational judgement, (d) awareness of the necessity to perform B-CPR, (e) initial position of the patient, (f) automaticity of actions. Integrating these concepts into dispatch protocols and layperson training may improve dispatcher-bystander interaction and the outcomes of 'at-home CA'.


[Mobile laboratory for autopsies and histological studies].
The laboratory is set up in an "Ikarus-556" bus and has two sections: histological laboratory and an autopsy room equipped with autonomous water supply and sewage systems, heating systems, and necessary medical equipment allowing one to perform autopsies of the fatal cases and histological examinations under field conditions, in district and rural hospitals, and to improve the quality of therapeutic and diagnostic work.


Antibiotic-induced dysbiosis of gut microbiota impairs corneal development in postnatal mice by affecting CCR2 negative macrophage distribution.
Antibiotics are extremely useful, but they can cause adverse impacts on host bodies. We found that antibiotic treatment altered the composition of the gut microbiota and the gene expression profile in the corneal tissues of postnatal mice and decreased the corneal size and thickness, the angiogenesis of limbal blood vessels, and the neurogenesis of corneal nerve fibers. The reconstitution of the gut microbiota with fecal transplants in antibiotic-treated mice largely reversed these impairments in corneal development. Furthermore, C-C chemokine receptor type 2 negative (CCR2-) macrophages were confirmed to participate in corneal development, and their distribution in the cornea was regulated by the gut microbiota. We propose that the CCR2- macrophage population is a crucial mediator through which gut microbiota affect corneal development in postnatal mice. In addition, probiotics were shown to have the potential effect of restoring corneal development in antibiotic-treated mice. Abx-induced gut dysbiosis has significant, long-term effects on the development of the cornea, and reversal of these suppressive effects takes a long time.


New method for the spin quantitation of [4Fe-4S](+) clusters with S = (3)/(2). Application to the FS0 center of the NarGHI nitrate reductase from Escherichia coli.
In conventional analyses of g approximately 5 signals given by [4Fe-4S](+) clusters with S = 3/2, the effective g values that cannot be measured in the electron paramagnetic resonance (EPR) spectrum are deduced from rhombograms calculated by assuming that the g matrix is isotropic with g(x) = g(y) = g(z) = 2.00. We have shown that when the two low-field peaks corresponding to the Kramers doublets are visible in the spectrum, a new, independent piece of information about the system can be obtained by studying the temperature dependence of the ratio of the area under these peaks. By applying this method to the g approximately 5 signals displayed by NarGHI nitrate reductase, we were able to determine all the parameters of the spin Hamiltonian of FS0 centers with S = 3/2 and to measure accurately their number. Our results indicate that simple analyses based on the assumption of an isotropic g matrix can give rise to very large errors.


An unconditionally stable numerical method for the Luo-Rudy 1 model used in simulations of defibrillation.
Numerical simulations of defibrillation using the Bidomain model coupled to a model of membrane kinetics represent a serious numerical challenge. This is because very high voltages close to defibrillation electrodes demand that extreme time step restrictions be placed on standard numerical schemes, e.g. the forward Euler scheme. A common solution to this problem is to modify the cell model by simple if-tests applied to several equations and rate functions. These changes are motivated by numerical problems rather than physiology, and should therefore be avoided whenever possible. The purpose of this paper is to present a numerical scheme that handles the original model without modifications and which is unconditionally stable for the Luo-Rudy phase 1 model. This also shows that the cell model is mathematically well-behaved, even in the presence of very high voltages. Our theoretical results are illustrated by numerical computations.


Manganese activation of superoxide dismutase 2 in the mitochondria of Saccharomyces cerevisiae.
Manganese-dependent superoxide dismutase 2 (SOD2) in the mitochondria plays a key role in protection against oxidative stress. Here we probed the pathway by which SOD2 acquires its manganese catalytic cofactor. We found that a mitochondrial localization is essential. A cytosolic version of Saccharomyces cerevisiae Sod2p is largely apo for manganese and is only efficiently activated when cells accumulate toxic levels of manganese. Furthermore, Candida albicans naturally produces a cytosolic manganese SOD (Ca SOD3), yet when expressed in the cytosol of S. cerevisiae, a large fraction of Ca SOD3 also remained manganese-deficient. The cytosol of S. cerevisae cannot readily support activation of Mn-SOD molecules. By monitoring the kinetics for metalation of S. cerevisiae Sod2p in vivo, we found that prefolded Sod2p in the mitochondria cannot be activated by manganese. Manganese insertion is only possible with a newly synthesized polypeptide. Furthermore, Sod2p synthesis appears closely coupled to Sod2p import. By reversibly blocking mitochondrial import in vivo, we noted that newly synthesized Sod2p can enter mitochondria but not a Sod2p polypeptide that was allowed to accumulate in the cytosol. We propose a model in which the insertion of manganese into eukaryotic SOD2 molecules is driven by the protein unfolding process associated with mitochondrial import.


Use of the interview in admissions.
A description of one method of dealing with nonacademic information from applicants seeking acceptance into a physical therapy curriculum is presented. The strengths of this admission process are interviewer training sessions, use of simulated interviews presented by videotape, subsequent discussion of ratings, specific criteria to support interview ratings, and an arbitration procedure. Attempts were made to increase the objectivity of the interview through frequent meetings of the admission committee to decide on the areas of the interview and specific criteria on which to rate each area. In the areas of weighting factors, training of interviewers, development of specific interview criteria, arbitration procedure, and the use of the computer printout, this admission process could be easily adapted to other allied health programs. Details of the interview are discussed.


Integrative Analysis of miRNA and inflammatory gene expression after acute particulate matter exposure.
MicroRNAs (miRNAs) are environmentally sensitive inhibitors of gene expression that may mediate the effects of metal-rich particulate matter (PM) and toxic metals on human individuals. Previous environmental miRNA studies have investigated a limited number of candidate miRNAs and have not yet evaluated the functional effects on gene expression. In this study, we wanted to identify PM-sensitive miRNAs using microarray profiling on matched baseline and postexposure RNA from foundry workers with well-characterized exposure to metal-rich PM and to characterize miRNA relations with expression of candidate inflammatory genes. We applied microarray analysis of 847 human miRNAs and real-time PCR analysis of 18 candidate inflammatory genes on matched blood samples collected from foundry workers at baseline and after 3 days of work (postexposure). We identified differentially expressed miRNAs (fold change [FC] > 2 and p < 0.05) and correlated their expression with the inflammatory associated genes. We performed in silico network analysis in MetaCore v6.9 to characterize the biological pathways connecting miRNA-mRNA pairs. Microarray analysis identified four miRNAs that were differentially expressed in postexposure compared with baseline samples, including miR-421 (FC = 2.81, p < 0.001), miR-146a (FC = 2.62, p = 0.007), miR-29a (FC = 2.91, p < 0.001), and let-7g (FC = 2.73, p = 0.019). Using false discovery date adjustment for multiple comparisons, we found 11 miRNA-mRNA correlated pairs involving the 4 differentially expressed miRNAs and candidate inflammatory genes. In silico network analysis with MetaCore database identified biological interactions for all the 11 miRNA-mRNA pairs, which ranged from direct mRNA targeting to complex interactions with multiple intermediates. Acute PM exposure may affect gene regulation through PM-responsive miRNAs that directly or indirectly control inflammatory gene expression.


SLC5A8 nuclear translocation and loss of expression are associated with poor outcome in pancreatic ductal adenocarcinoma.
This study aimed to assess the role of SLC5A8 expression in the survival of pancreatic cancer. We determined SLC5A8 expression in pancreatic ductal adenocarcinoma and adjacent non-neoplastic pancreas (NNP) obtained from 110 patients who underwent pancreatectomy. Formalin-fixed paraffin-embedded core sections in a tissue microarray were immunostained using polyclonal anti-SLC5A8 antibody, and a semiquantitative measure of SLC5A8 expression was determined. SLC5A8 expression was low in 56% (62/110) of pancreatic cancers as compared to NNP that had low expression in only 9% (10/107) of specimens (P < 0.0001). All cells expressing SLC5A8 did so in the cytoplasm, whether they are neoplastic or not. Nuclear expression of SLC5A8 occurred in 38% (42/110) of cancers, but it was uncommon in NNP (7%, 8/107) (P < 0.0001). Kaplan-Meier estimates showed that survival in patients whose cancers had low SLC5A8 expression, and/or nuclear expression, was significantly worse than in patients whose cancers had none of these abnormalities (P = 0.02). For the 88 patients whose cancers had abnormal SLC5A8 expression, median survival was 1.4 years, as compared to 3.9 years in patients whose cancers both expressed high levels of SLC5A8 and lacked nuclear expression. SLC5A8 nuclear translocation and loss of expression are associated with poor outcome in pancreatic ductal adenocarcinoma.


Removal of Uranium From Aqueous Solution by Carbon Nanotubes.
The adsorption of uranium onto carbon nanotubes (CNTs) was investigated. The effect of solution pH, contact time, initial uranium concentration, and temperature on the adsorption capacity of uranium was determined. CNTs were characterized by using scanning electron microscopy (SEM), transmission electron microscopy (TEM), powder x-ray diffraction (XRD), Raman spectra, and the Fourier infrared spectra (FTIR). The diameters of the CNTs varied from 10 to 50 nm in diameter and 1 ~ 2 μm in length. FTIR spectra analysis indicated that carboxyl groups were involved in adsorption of U(VI) by CNTs. The experimental results showed that U(VI) adsorption onto CNTs reached equilibrium within 10 min, and the removal efficiency was 95% at pH = 5. The adsorption kinetics of U(VI) could be described by a pseudo first-order kinetic model. The adsorption isotherm conformed to the Slips model. The adsorption process was spontaneous and endothermic.


Helical CT angiography of fenestrated stent grafting of abdominal aortic aneurysms.
Fenestrated stent grafts have been developed to treat patients with abdominal aortic aneurysms (AAA) associated with complicated aneurysm necks, such as short necks, severe angulated or poor quality necks (presence of calcification or thrombus). The technique is performed by creating an opening in the graft material so that the stent graft can be placed above the renal and other visceral branches without compromising blood perfusion to these vessels. In most situations, a supporting stent is inserted into the fenestrated vessel to provide fixation of the fenestrated vessel against stent grafts, as well as to preserve patency of the vessel. Helical CT angiography (CTA) is the preferred imaging modality in both pre-operative planning and post-procedural follow-up of fenestrated repair of AAA. The main concerns of fenestrated stent grafting lie in the following two aspects: patency of the fenestrated vessels and position of the fenestrated stents in relation to the artery branches. In this article, the author presents the clinical applications of 2D and 3D visualizations in the follow-up of patients with AAA treated with fenestrated stent grafts, with the aim of providing useful information to readers and increasing their knowledge of an increasingly used technique, fenestrated stent grafting in the treatment of AAA.


The Egyptian sycamore tree: its use in dermatology.
An extract of the sycamore tree, known by the ancient Egyptians for its medicinal properties, was used to treat certain dermatoses in a group of Egyptian patients. Twenty-one patients having a variety of conditions including pityriasis alba, atopic dermatosis, psoriasis and tina infections were given a cream containing the active material. The response was excellent.


An overview and methodological assessment of systematic reviews and meta-analyses of enhanced recovery programmes in colorectal surgery.
To identify and critically assess the extent to which systematic reviews of enhanced recovery programmes for patients undergoing colorectal surgery differ in their methodology and reported estimates of effect. Review of published systematic reviews. We searched the Cochrane Database of Systematic Reviews, the Database of Abstracts of Reviews of Effects (DARE) and Health Technology Assessment (HTA) Database from 1990 to March 2013. Systematic reviews of enhanced recovery programmes for patients undergoing colorectal surgery were eligible for inclusion. The primary outcome was length of hospital stay. We assessed changes in pooled estimates of treatment effect over time and how these might have been influenced by decisions taken by researchers as well as by the availability of new trials. The quality of systematic reviews was assessed using the Centre for Reviews and Dissemination (CRD) DARE critical appraisal process. 10 systematic reviews were included. Systematic reviews of randomised controlled trials have consistently shown a reduction in length of hospital stay with enhanced recovery compared with traditional care. The estimated effect tended to increase from 2006 to 2010 as more trials were published but has not altered significantly in the most recent review, despite the inclusion of several unique trials. The best estimate appears to be an average reduction of around 2.5 days in primary postoperative length of stay. Differences between reviews reflected differences in interpretation of inclusion criteria, searching and analytical methods or software. Systematic reviews of enhanced recovery programmes show a high level of research waste, with multiple reviews covering identical or very similar groups of trials. Where multiple reviews exist on a topic, interpretation may require careful attention to apparently minor differences between reviews. Researchers can help readers by acknowledging existing reviews and through clear reporting of key decisions, especially on inclusion/exclusion and on statistical pooling.


Semisolid meal enriched in oat bran decreases plasma glucose and insulin levels, but does not change gastrointestinal peptide responses or short-term appetite in healthy subjects.
Dietary fibre (DF) may play an important role in weight control. The amount, type and way of processing of DF modify food structure and subsequent postprandial appetitive, metabolic and hormonal effects, but current understanding about the magnitude of effects that specific types and amounts of DF exert are still poorly understood. We investigated the effects of wheat and oat brans alone and as combination in semisolid food matrix on postprandial appetite profile and gastrointestinal (GI) hormonal responses. Twenty healthy, normal-weight subjects (5 male/15 female, aged 23.3 ± 0.85y) participated in the study. Isoenergetic and isovolumic (1250 kJ, 300 g) puddings with different insoluble and soluble DF content were tested in a randomised order: pudding with 1) no added fibre, 2) 10 g wheat bran DF, 3) 10 g oat bran DF and 4) combination including 5 g wheat bran DF + 5 g oat bran DF. Blood samples were drawn before and 15, 30, 45, 60, 90, 120 and 180 min after the test meals to determine plasma glucose, ghrelin, peptide YY (PYY) and serum insulin concentrations. Subjective profiles of appetite were assessed using visual analogue scales (VAS). Plasma glucose (P = 0.001) and serum insulin (P < 0.001) responses were the lowest after the pudding with the greatest amount of β-glucan. In contrast, postprandial ghrelin or PYY responses or appetite sensations did not differ among the meals. Oat β-glucan decreased postprandial plasma glucose and serum insulin responses, yet had no significant effects on GI peptide responses or appetite ratings.


Factors influencing laboratory animal spontaneous tumor profiles.
In chemical carcinogenicity and drug-safety testing, a carcinogen is defined as an agent that when administered by an appropriate route causes an increased incidence of tumors in experimental animals as compared to unexposed control animals. Although a carcinogen may cause the appearance of tumors in organs where tumors do not usually occur in a given strain, the usual response is to increase the types of tumors seen spontaneously and to shorten the period of latency. The use of carcinogenesis experiments for research and safety assessment requires properly designed and well-conducted experiments and a knowledge of background data and variations in tumor incidences of control animals. Many factors can influence the reported incidences of spontaneous tumors. These include species, strain, sex, age, and source of the experimental test animal; study duration; extent of the pathology examination; dietary and environmental conditions; qualifications and experience of the study pathologist; diagnostic criteria and nomenclature conventions; and quality assurance and review procedures. This paper discusses several factors which may influence the incidence of tumors in control and test animals, and provides examples to illustrate the potential for these factors to affect the data.


Safety and efficacy of a recombinant hepatitis E vaccine.
Hepatitis E virus (HEV) is an important cause of viral hepatitis. We evaluated the safety and efficacy of an HEV recombinant protein (rHEV) vaccine in a phase 2, randomized, double-blind, placebo-controlled trial. In Nepal, we studied 2000 healthy adults susceptible to HEV infection who were randomly assigned to receive three doses of either the rHEV vaccine or placebo at months 0, 1, and 6. Active (including hospital) surveillance was used to identify acute hepatitis and adverse events. The primary end point was the development of hepatitis E after three vaccine doses. A total of 1794 subjects (898 in the vaccine group and 896 in the placebo group) received three vaccine doses; the total vaccinated cohort was followed for a median of 804 days. After three vaccine doses, hepatitis E developed in 69 subjects, of whom 66 were in the placebo group. The vaccine efficacy was 95.5% (95% confidence interval [CI], 85.6 to 98.6). In an intention-to-treat analysis that included all 87 subjects in whom hepatitis E developed after the first vaccine dose, 9 subjects were in the vaccine group, with a vaccine efficacy of 88.5% (95% CI, 77.1 to 94.2). Among subjects in a subgroup randomly selected for analysis of injection-site findings and general symptoms (reactogenicity subgroup) during the 8-day period after the administration of any dose, the proportion of subjects with adverse events was similar in the two study groups, except that injection-site pain was increased in the vaccine group (P=0.03). In a high-risk population, the rHEV vaccine was effective in the prevention of hepatitis E. (ClinicalTrials.gov number, NCT00287469 [ClinicalTrials.gov].).


Pyrrolidine dithiocarbamate protects against scopolamine-induced cognitive impairment in rats.
Alzheimer's disease (AD) is a chronic neurodegenerative disorder that leads to disturbances of cognitive functions. Although the primary cause of AD remains unclear, brain acetylcholine deficiency, oxidative stress and neuroinflammation may be considered the principal pathogenic factors. The present study was constructed to investigate the anti-amnestic effect of pyrrolidine dithiocarbamate (PDTC) on scopolamine-induced behavioral, neurochemical and biochemical changes in rats. PDTC (50 and 100mg/kg) and donepezil (2.5mg/kg) were orally administered for 14 successive days. Dementia was induced at the end of the treatment period by a single injection of scopolamine (20mg/kg; i.p.), and Y-maze test was conducted 30min thereafter. Rats were then sacrificed and homogenates of cortical and hippocampal tissues were used for the estimation of noradrenaline, dopamine, serotonin and heat shock protein 70 contents along with acetylcholinesterase activity. In addition, certain oxidative stress markers, pro-inflammatory and anti-inflammatory cytokines were assessed. Histological examination of cortical and hippocampal tissues was also performed. Scopolamine resulted in memory impairment that was coupled by alterations in the estimated neurotransmitters, heat shock protein 70, acetylcholinesterase activity, oxidative stress as well as inflammatory biomarkers. Histological analysis revealed serious damaging effects of scopolamine on the structure of cerebral cortex and hippocampus. Pretreatment of rats with PDTC in both doses mitigated scopolamine-induced behavioral, biochemical, neurochemical and histological changes in a manner comparable to donepezil. The observed anti-amnestic effect of PDTC makes it a promising candidate for clinical trials in patients with cognitive impairment.


The control of anti-donor immune responses by regulatory T cells in organ transplant patients.
The role of regulatory T cells in the induction and maintenance of transplant tolerance has been widely investigated in experimental animal models. Their involvement in the regulation of allogeneic immune reactivity in immunosuppressed organ transplant patients, however, remains unclear. Measurements of regulatory T cells after clinical organ transplantation may contribute to understanding their role in anti-donor immune responses. Several studies have investigated the frequency and/or immune regulatory function of peripheral regulatory T-cell populations, such as, the CD4+CD25+ regulatory T cells or the CD8+CD28- suppressor T cells, in clinically stable organ transplant patients and patients with acute or chronic rejection. This review summarizes these studies and discusses the correlations found between the number and function of regulatory T cells and the immunological state of the patient. This knowledge is warranted for a better understanding of the control of allogeneic immune responses by regulatory T cells. Subsequently, monitoring regulatory T cells may help us to identify patients in whom the anti-donor reactivity is actively suppressed.


Prognostic factors and risk stratification in patients with castration-resistant prostate cancer receiving docetaxel-based chemotherapy.
While novel drugs have been developed, docetaxel remains one of the standard initial systemic therapies for castration-resistant prostate cancer (CRPC) patients. Despite the excellent anti-tumor effect of docetaxel, its severe adverse effects sometimes distress patients. Therefore, it would be very helpful to predict the efficacy of docetaxel before treatment. The aims of this study were to evaluate the potential value of patient characteristics in predicting overall survival (OS) and to develop a risk classification for CRPC patients treated with docetaxel-based chemotherapy. This study included 79 patients with CRPC treated with docetaxel. The variables, including patient characteristics at diagnosis and at the start of chemotherapy, were retrospectively collected. Prognostic factors predicting OS were analyzed using the Cox proportional hazard model. Risk stratification for overall survival was determined based on the results of multivariate analysis. PSA response ≥50 % was observed in 55 (69.6 %) of all patients, and the median OS was 22.5 months. The multivariate analysis showed that age, serum PSA level at the start of chemotherapy, and Hb were independent prognostic factors for OS. In addition, ECOG performance status (PS) and the CRP-to-albumin ratio were not significant but were considered possible predictors for OS. Risk stratification according to the number of these risk factors could effectively stratify CRPC patients treated with docetaxel in terms of OS. Age, serum PSA level at the start of chemotherapy, and Hb were identified as independent prognostic factors of OS. ECOG PS and the CRP-to-albumin ratio were not significant, but were considered possible predictors for OS in Japanese CRPC patients treated with docetaxel. Risk stratification based on these factors could be helpful for estimating overall survival.


Nitrogen isotope composition of organically and conventionally grown crops.
Authentic samples of commercially produced organic and conventionally grown tomatoes, lettuces, and carrots were collected and analyzed for their delta15N composition in order to assemble datasets to establish if there are any systematic differences in nitrogen isotope composition due to the method of production. The tomato and lettuce datasets suggest that the different types of fertilizer commonly used in organic and conventional systems result in differences in the nitrogen isotope composition of these crops. A mean delta15N value of 8.1 per thousand was found for the organically grown tomatoes compared with a mean value of -0.1 per thousand for those grown conventionally. The organically grown lettuces had a mean value of 7.6 per thousand compared with a mean value of 2.9 per thousand for the conventionally grown lettuces. The mean value for organic carrots was not significantly different from the mean value for those grown conventionally. Overlap between the delta15N values of the organic and conventional datasets (for both tomatoes and lettuces) means that it is necessary to employ a statistical methodology to try and classify a randomly analyzed "off the shelf" sample as organic/conventional, and such an approach is demonstrated. Overall, the study suggests that nitrogen isotope analysis could be used to provide useful "intelligence" to help detect the substitution of certain organic crop types with their conventional counterparts. However, delta15N analysis of a "test sample" will not provide unequivocal evidence as to whether synthetic fertilizers have been used on the crop but could, for example, in a situation when there is suspicion that mislabeling of conventionally grown crops as "organic" is occurring, be used to provide supporting evidence.


Recommendations for SARS-CoV-2/COVID-19 testing: a scoping review of current guidance.
Testing used in screening, diagnosis and follow-up of COVID-19 has been a subject of debate. Several organisations have developed formal advice about testing for COVID-19 to assist in the control of the disease. We collated, delineated and appraised current worldwide recommendations about the role and applications of tests to control SARS-CoV-2/COVID-19. We searched for documents providing recommendations for COVID-19 testing in PubMed, EMBASE, LILACS, the Coronavirus Open Access Project living evidence database and relevant websites such as TRIP database, ECRI Guidelines Trust, the GIN database, from inception to 21 September 2020. Two reviewers applied the eligibility criteria to potentially relevant citations without language or geographical restrictions. We extracted data in duplicate, including assessment of methodological quality using the Appraisal of Guidelines for Research and Evaluation-II tool. We included 47 relevant documents and 327 recommendations about testing. Regarding the quality of the documents, we found that the domains with the lowest scores were 'Editorial independence' (Median=4%) and 'Applicability' (Median=6%). Only six documents obtained at least 50% score for the 'Rigour of development' domain. An important number of recommendations focused on the diagnosis of suspected cases (48%) and deisolation measures (11%). The most frequently recommended test was the reverse transcription-PCR (RT-PCR) assay (87 recommendations) and the chest CT (38 recommendations). There were 22 areas of agreement among guidance developers, including the use of RT-PCR for SARS-Cov-2 confirmation, the limited role of bronchoscopy, the use chest CT and chest X-rays for grading severity and the co-assessment for other respiratory pathogens. This first scoping review of recommendations for COVID-19 testing showed many limitations in the methodological quality of included guidance documents that could affect the confidence of clinicians in their implementation. Future guidance documents should incorporate a minimum set of key methodological characteristics to enhance their applicability for decision making.


Impact of viral and host DNA methylations on HPV16-related cervical cancer pathogenesis.
Epigenetic alterations within human papillomavirus (HPV) and host cellular genomes are known to occur during cervical carcinogenesis. Our objective was to analyse the influence of (1) methylation within two immunostimulatory CpG motifs within HPV16 E6 and E7 genes around the viral late promoter and their correlation, if any, with expression deregulation of host receptor (TLR9) and DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) and (2) global DNA methylation levels within CpGs of the repetitive Alu sequences, on cervical cancer (CaCx) pathogenesis. Significantly higher proportions of CaCx samples portrayed methylation in immunostimulatory CpG motifs, compared to HPV16-positive non-malignant samples, with cases harbouring episomal HPV16 showing decreased methylation compared to those with viral integration. A significant linear trend of TLR9 upregulation was recorded in the order of HPV-negative controls < HPV16-positive non-malignant samples < HPV16-positive CaCx cases. TLR9 upregulation in cases with episomal HPV16 was again higher among those with non-methylated immunostimulatory CpG motifs. Comparison of cases with HPV-negative controls revealed that DNMT3A was significantly downregulated only among integrated cases, DNMT3B was significantly overexpressed among both categories of cases, although at variable levels, while DNMT1 failed to show any deregulated expression among the cases. Global host DNA hypomethylation, also showed a significant linear increasing trend through the progressive CaCx development stages mentioned above and was most prominently higher among cases with episomal HPV16 as opposed to viral integration. Thus, HPV16 and host methylations appear to influence CaCx pathogenesis, with differential molecular signatures among CaCx cases with episomal and integrated HPV16.


Effects of acute resistance exercise with different loads on appetite, appetite hormones and autonomic nervous system responses in healthy young men.
Although the effect of continuous aerobic exercise on the appetite has been widely explored, the influence of resistance exercise (RE) with different variables, including training loads, training volume, and inter-set rest, on appetite responses requires further investigation. This study examined the importance of training load in RE-induced appetite regulation, with the total training volume and inter-set rest equalized. In total, 11 healthy young men (age = 23 ± 2 years, body mass index = 22 ± 2 kg/m2) were included. Participants completed 3 trials, namely moderate-load RE (MOD; 4 sets of 8 repetitions at 85% 8RM), low-load RE (LOW; 4 sets of 15 repetitions at 45% 8RM), and a control (CON; no exercise), in a randomized, crossover design. Subjective appetite ratings; concentrations of ghrelin, peptide YY (PYY), and lactate; and the autonomic nervous system activity were evaluated before exercise and 1 h after exercise. The hunger and predicted food consumption ratings, and ghrelin concentrations immediately after exercise were significantly lower in the MOD and LOW trials (p < 0.05 vs. CON). The PYY and lactate concentrations immediately after exercise were significantly higher in the MOD and LOW trials (p < 0.05 vs. CON). Heart rate variability recovery was slower in the MOD trial. These findings suggest that both moderate-load and low-load RE at equal training volumes and inter-set rest induce similar responses on hunger suppression and orexigenic signals, except for the slower recovery of autonomic modulation after moderate-load RE. Our results suggest that when individuals aim to potentiate appetite suppression after a bout of RE, both moderate- and low-load RE could be applied.


Post-processing of structural MRI for individualized diagnostics.
Currently, a relevant proportion of all histopathologically proven focal cortical dysplasia (FCD) escape visual detection; this shows the need for additional improvements in analyzing MRI data. A positive MRI is still the strongest prognostic factor for postoperative freedom of seizures. Among several post-processing methods voxel-based morphometry (VBM) of T1- and T2-weighted sequences and T2 relaxometry are routinely applied in pre-surgical diagnostics of cryptogenic epilepsy in epilepsy centers. VBM is superior to conventional visual analysis with 9-15% more identified epileptogenic foci, while T2 relaxometry has its main application in (mesial) temporal lobe epilepsy. Further methods such as surface-based morphometry (SBM) or diffusion tensor imaging are promising but there is a lack of current studies comparing their individual diagnostic value. Post-processing methods represent an important addition to conventional visual analysis but need to be interpreted with expertise and experience so that they should be apprehended as a complementary tool within the context of the multi-modal evaluation of epilepsy patients. This review will give an overview of existing post-processing methods of structural MRI and outline their clinical relevance in detection of epileptogenic structural changes.


Behavioral and emotional problems among Italian international adoptees and non-adopted children: father's and mother's reports.
This study intends to fill the gap in empirical research carried out in Italy regarding international adoptees' behavioral and emotional problems. Assuming a multi-informant approach, it aims to compare parents' reports of behavioral problems of adopted and non-adopted children and to examine parental agreement. The sample was composed of 186 adoptive couples and 195 biological couples with the target child between ages 7 and 11 years. The mother and father filled in the Child Behavior Checklist (CBCL) separately. Analysis of the CBCL revealed that adopted children are perceived by their parents as having more Total and Externalizing Problems than are their non-adopted counterparts. Moreover, they are more likely to demonstrate attention difficulties and aggressive behavior. The agreement between parents turned out to be moderate for adoptive parents and slightly lower for the biological ones. Consistent with most adoption research, the results confirm the higher risk of behavioral problems among adopted children. They also shed light on the significant perceptual discrepancy between mothers and fathers, underlining the importance of considering both parents' reports in the study of adopted children's adjustment.


Expression of p16 in conjunctival intraepithelial neoplasia does not correlate with HPV-infection.
The aim of our study was to identify the frequency of expression of p16(INK4a) (CDKN2A) and HPV (human papilloma virus) in different grades of conjunctival intraepithelial neoplasia (CIN).Twelve specimens including CIN I (2), II (3), III (5), and CIN with beginning invasion (2), as well as 15 control specimens, were stained with antibodies against p16(INK4a) and MIB1. The presence of HPV was examined by PCR.p16 as well as MIB1 were significantly elevated in CIN compared to control specimens (p<0.01) without correlation with the differentiation grade. Only two cases with CIN grade 3 contained HPV 16.As few control specimens also showed increased p16(INK4a) expression, p16(INK4a) seems not to be a very reliable marker for the exact determination of CIN. It could serve as an additional diagnostic tool besides the morphological characterization. Our study suggested that p16(INK4a) elevation is not associated with HPV infection.


Self-Assembled Micelles of Amphiphilic PEGylated Drugs for Cancer Treatment.
Generally, poor solubility and imprecise delivery of chemotherapeutic drugs can compromise their efficacies for clinical cancer treatment. In order to address such concerns, poor water-soluble drugs are conjugated with poly(ethylene glycol) (PEG) to obtain PEGylated drugs, which have improved water solubility and can also self-assemble in an aqueous solution to form micelles (PEGylated drug micelles). The surface PEG layer enhances the micelles' colloidal stability and reduces the interaction with physiological surroundings. Meanwhile, PEGylated drug micelles are tumor- targeting via the enhanced permeation and retention (EPR) effect to improve antitumor efficacy in comparison with free drugs. PEGylated drug micelles employ drugs as parts of the carrier medium, which increases the micelles' drug loading capacity relatively. The development of stimuli- responsive PEGylated drug micelles facilitates the drug release to be smart and controllable. Moreover, the PEGylated drug micelles show great potentials in overcoming the challenges of cancer therapy, such as multidrug resistance (MDR), angiogenesis, immunosuppression, and so on. In this review, we highlight the research progresses of PEGylated drug micelles, including the structures and properties, smart stimuli-responsive PEGylated drug micelles, and the challenges that have been overcome by PEGylated drug micelles.


Nocturnal light exposure alters hepatic Pai-1 expression by stimulating the adrenal pathway in C3H mice.
Recent studies have suggested the possibility that nocturnal light exposure affects many biological processes in rodents, especially the circadian rhythm, an endogenous oscillation of approximately 24 h. However, there is still insufficient information about the physiological effects of nocturnal light exposure. In this study, we examined the changes in gene expression and serum levels of plasminogen activator inhibitor-1 (PAI-1), a major component of the fibrinolytic system that shows typical circadian rhythmicity, in C3H/He mice. Zeitgeber time (ZT) was assessed with reference to the onset of light period (ZT0). Exposure to fluorescent light (70 lux) for 1 h in the dark period (ZT14) caused a significant increase in hepatic Pai-1 gene expression at ZT16. Serum PAI-1 levels also tended to increase, albeit not significantly. Expression levels of the typical clock genes Bmal1, Clock, and Per1 were significantly increased at ZT21, ZT16, and ZT18, respectively. Exposure to nocturnal light significantly increased plasma adrenalin levels. The effects of nocturnal light exposure on Pai-1 expression disappeared in adrenalectomized mice, although the changes in clock genes were still apparent. In conclusion, our results suggest that nocturnal light exposure, even for 1 h, alters hepatic Pai-1 gene expression by stimulating the adrenal pathway. Adrenalin secreted from the adrenal gland may be an important signaling mediator of the change in Pai-1 expression in response to nocturnal light exposure.


Kinematics of 90 degrees running turns in wild mice.
Turning is a requirement for locomotion on the variable terrain that most terrestrial animals inhabit and is a deciding factor in many predator-prey interactions. Despite this, the kinematics and mechanics of quadrupedal turns are not well understood. To gain insight to the turning kinematics of small quadrupedal mammals, six adult wild mice were videotaped at 250 Hz from below as they performed 90 degrees running turns. Four markers placed along the sagittal axis were digitized to allow observation of lateral bending and body rotation throughout the turn. Ground contact periods of the fore- and hindlimbs were also noted for each frame. During turning, mice increased their ground contact time, but did not change their stride frequency relative to straight running at maximum speed. Postcranial body rotation preceded deflection in heading, and did not occur in one continuous motion, but rather in bouts of 15-53 degrees. These bouts were synchronized with the stride cycle, such that the majority of rotation occurred during the second half of forelimb support and the first half of hindlimb support. In this phase of the stride cycle, the trunk was sagittally flexed and rotational inertia was 65% of that during maximal extension. By synchronizing body rotation with this portion of the stride cycle, mice can achieve a given angular acceleration with much lower applied torque. Compared with humans running along curved trajectories, mice maintained relatively higher speeds at proportionately smaller radii. A possible explanation for this difference lies in the more crouched limb posture of mice, which increases the mechanical advantage for horizontal ground force production. The occurrence of body rotation prior to deflection in heading may facilitate acceleration in the new direction by making use of the relatively greater force production inherent in the parasagittal limb posture of mice.


Cancer stem cells in hepatocellular carcinoma: an overview and promising therapeutic strategies.
The poor clinical outcome of hepatocellular carcinoma (HCC) patients is ascribed to the resistance of HCC cells to traditional treatments and tumor recurrence after curative therapies. Cancer stem cells (CSCs) have been identified as a small subset of cancer cells which have high capacity for self-renewal, differentiation and tumorigenesis. Recent advances in the field of liver CSCs (LCSCs) have enabled the identification of CSC surface markers and the isolation of CSC subpopulations from HCC cells. Given their central role in cancer initiation, metastasis, recurrence and therapeutic resistance, LCSCs constitute a therapeutic opportunity to achieve cure and prevent relapse of HCC. Thus, it is necessary to develop therapeutic strategies to selectively and efficiently target LCSCs. Small molecular inhibitors targeting the core stemness signaling pathways have been actively pursued and evaluated in preclinical and clinical studies. Other alternative therapeutic strategies include targeting LCSC surface markers, interrupting the CSC microenvironment, and altering the epigenetic state. In this review, we summarize the properties of CSCs in HCC and discuss novel therapeutic strategies that can be used to target LCSCs.


Phylogeny and regulation of four lipocalin genes clustered in the chicken genome: evidence of a functional diversification after gene duplication.
A novel lipocalin gene is here reported that represents the fourth member of a cluster we have identified in the chicken genome. This cluster also includes Chondrogenesis-Associated Lipocalins beta and gamma (CAL beta, CAL gamma) and Extracellular Fatty Acid Binding Protein (Ex-FABP). The new gene codes for a 22-kDa secreted protein with three cysteine residues and a series of sequence features well conserved in the lipocalin family. All the genes in the cluster are structurally similar presenting comparable exon/intron boundary positions and exon sizes. A phylogenetic analysis indicates the monophyletic grouping of these genes, and their relationship with the lipocalins alpha-1-microglobulin (A1mg), complement factor 8 gamma chain (C8GC), prostaglandin D synthase (PGDS), and neutrophil-gelatinase-associated lipocalin (NGAL). The new cluster gene appears to be the ortholog of the mammalian C8GC and was thus named Ggal-C8GC. This orthology also suggests that this lipocalin was present in the ancestor common to reptiles and mammals. In addition to other expressing tissues, Ex-FABP, CAL beta and CAL gamma genes are highly transcribed in chondrocytes at late stages of chondrogenesis during endochondral bone formation and/or upon inflammatory stimulation. Here, we show that they are also transcriptionally induced when chondrocytes are subjected to various biological events as cell quiescence, cell shape transition, and hormonal stimulation. By contrast, Ggal-C8GC transcripts are only barely detectable in chondrocytes, but are more abundant in liver, kidney, brain, heart, skeletal muscle and particularly in skin. Moreover, no expression induction was observed neither during chondrocyte differentiation, nor upon any of the stimulations mentioned above. This indicates that the Ggal-C8GC gene was co-opted for a novel function after the duplication events that gave rise to the cluster. The peculiar coordinated regulation of Ex-FABP, CAL beta and CAL gamma, and the apparent divergent role of Ggal-C8GC suggest that these gene duplications may have been maintained during evolution by a sub-functionalization mechanism where some common function(s) are shared by several members of the cluster and some other specialized function(s) are unique to other members.


Size dependent fragmentation of argon clusters in the soft x-ray ionization regime.
Photofragmentation of argon clusters of average size ranging from 10 up to 1000 atoms is studied using soft x-ray radiation below the 2p threshold and multicoincidence mass spectroscopy technique. For small clusters (<N>=10), ionization induces fast fragmentation with neutral emission imparting a large amount of energy. While the primary dissociation takes place on a picosecond time scale, the fragments undergo slow degradation in the spectrometer on a microsecond time scale. For larger clusters (<N> >or=100) we believe that we observe the fragmentation pattern of multiply charged species on a time-scale which lasts a few hundred nanoseconds. The reason for these slower processes is the large number of neutral atoms which act as an efficient cooling bath where the excess energy ("heat") dissipates among all degrees of freedom. Further degradation of the photoionic cluster in spectrometer then takes place on the microsecond time scale, similar to small clusters.


Predicting the occurrence of embolic events: an analysis of 1456 episodes of infective endocarditis from the Italian Study on Endocarditis (SEI).
Embolic events are a major cause of morbidity and mortality in patients with infective endocarditis. We analyzed the database of the prospective cohort study SEI in order to identify factors associated with the occurrence of embolic events and to develop a scoring system for the assessment of the risk of embolism. We retrospectively analyzed 1456 episodes of infective endocarditis from the multicenter study SEI. Predictors of embolism were identified. Risk factors identified at multivariate analysis as predictive of embolism in left-sided endocarditis, were used for the development of a risk score: 1 point was assigned to each risk factor (total risk score range: minimum 0 points; maximum 2 points). Three categories were defined by the score: low (0 points), intermediate (1 point), or high risk (2 points); the probability of embolic events per risk category was calculated for each day on treatment (day 0 through day 30). There were 499 episodes of infective endocarditis (34%) that were complicated by ≥ 1 embolic event. Most embolic events occurred early in the clinical course (first week of therapy: 15.5 episodes per 1000 patient days; second week: 3.7 episodes per 1000 patient days). In the total cohort, the factors associated with the occurrence of embolism at multivariate analysis were prosthetic valve localization (odds ratio, 1.84), right-sided endocarditis (odds ratio, 3.93), Staphylococcus aureus etiology (odds ratio, 2.23) and vegetation size ≥ 13 mm (odds ratio, 1.86). In left-sided endocarditis, Staphylococcus aureus etiology (odds ratio, 2.1) and vegetation size ≥ 13 mm (odds ratio, 2.1) were independently associated with embolic events; the 30-day cumulative incidence of embolism varied with risk score category (low risk, 12%; intermediate risk, 25%; high risk, 38%; p < 0.001). Staphylococcus aureus etiology and vegetation size are associated with an increased risk of embolism. In left-sided endocarditis, a simple scoring system, which combines etiology and vegetation size with time on antimicrobials, might contribute to a better assessment of the risk of embolism, and to a more individualized analysis of indications and contraindications for early surgery.


Novel adverse outcome pathways revealed by chemical genetics in a developing marine fish.
Crude oil spills are a worldwide ocean conservation threat. Fish are particularly vulnerable to the oiling of spawning habitats, and crude oil causes severe abnormalities in embryos and larvae. However, the underlying mechanisms for these developmental defects are not well understood. Here, we explore the transcriptional basis for four discrete crude oil injury phenotypes in the early life stages of the commercially important Atlantic haddock (Melanogrammus aeglefinus). These include defects in (1) cardiac form and function, (2) craniofacial development, (3) ionoregulation and fluid balance, and (4) cholesterol synthesis and homeostasis. Our findings suggest a key role for intracellular calcium cycling and excitation-transcription coupling in the dysregulation of heart and jaw morphogenesis. Moreover, the disruption of ionoregulatory pathways sheds new light on buoyancy control in marine fish embryos. Overall, our chemical-genetic approach identifies initiating events for distinct adverse outcome pathways and novel roles for individual genes in fundamental developmental processes.


Effect of school based physical activity programme (KISS) on fitness and adiposity in primary schoolchildren: cluster randomised controlled trial.
To assess the effectiveness of a school based physical activity programme during one school year on physical and psychological health in young schoolchildren. Cluster randomised controlled trial. 28 classes from 15 elementary schools in Switzerland randomly selected and assigned in a 4:3 ratio to an intervention (n=16) or control arm (n=12) after stratification for grade (first and fifth grade), from August 2005 to June 2006. 540 children, of whom 502 consented and presented at baseline. Children in the intervention arm (n=297) received a multi-component physical activity programme that included structuring the three existing physical education lessons each week and adding two additional lessons a week, daily short activity breaks, and physical activity homework. Children (n=205) and parents in the control group were not informed of an intervention group. For most outcome measures, the assessors were blinded. Primary outcome measures included body fat (sum of four skinfolds), aerobic fitness (shuttle run test), physical activity (accelerometry), and quality of life (questionnaires). Secondary outcome measures included body mass index and cardiovascular risk score (average z score of waist circumference, mean blood pressure, blood glucose, inverted high density lipoprotein cholesterol, and triglycerides). 498 children completed the baseline and follow-up assessments (mean age 6.9 (SD 0.3) years for first grade, 11.1 (0.5) years for fifth grade). After adjustment for grade, sex, baseline values, and clustering within classes, children in the intervention arm compared with controls showed more negative changes in the z score of the sum of four skinfolds (-0.12, 95 % confidence interval -0.21 to -0.03; P=0.009). Likewise, their z scores for aerobic fitness increased more favourably (0.17, 0.01 to 0.32; P=0.04), as did those for moderate-vigorous physical activity in school (1.19, 0.78 to 1.60; P<0.001), all day moderate-vigorous physical activity (0.44, 0.05 to 0.82; P=0.03), and total physical activity in school (0.92, 0.35 to 1.50; P=0.003). Z scores for overall daily physical activity (0.21, -0.21 to 0.63) and physical quality of life (0.42, -1.23 to 2.06) as well as psychological quality of life (0.59, -0.85 to 2.03) did not change significantly. A school based multi-component physical activity intervention including compulsory elements improved physical activity and fitness and reduced adiposity in children. Trial registration Current Controlled Trials ISRCTN15360785.


Multi-domain cognitive assessment of male mice shows space radiation is not harmful to high-level cognition and actually improves pattern separation.
Astronauts on interplanetary missions - such as to Mars - will be exposed to space radiation, a spectrum of highly-charged, fast-moving particles that includes 56Fe and 28Si. Earth-based preclinical studies show space radiation decreases rodent performance in low- and some high-level cognitive tasks. Given astronaut use of touchscreen platforms during training and space flight and given the ability of rodent touchscreen tasks to assess functional integrity of brain circuits and multiple cognitive domains in a non-aversive way, here we exposed 6-month-old C57BL/6J male mice to whole-body space radiation and subsequently assessed them on a touchscreen battery. Relative to Sham treatment, 56Fe irradiation did not overtly change performance on tasks of visual discrimination, reversal learning, rule-based, or object-spatial paired associates learning, suggesting preserved functional integrity of supporting brain circuits. Surprisingly, 56Fe irradiation improved performance on a dentate gyrus-reliant pattern separation task; irradiated mice learned faster and were more accurate than controls. Improved pattern separation performance did not appear to be touchscreen-, radiation particle-, or neurogenesis-dependent, as 56Fe and 28Si irradiation led to faster context discrimination in a non-touchscreen task and 56Fe decreased new dentate gyrus neurons relative to Sham. These data urge revisitation of the broadly-held view that space radiation is detrimental to cognition.


Effects of telmisartan on fat distribution: a meta-analysis of randomized controlled trials.
Several meta-analyses have confirmed the positive metabolic effects of telmisartan, an angiotensin II receptor blocker that can also act as a partial peroxisome proliferator-activated receptor-γ agonist, compared to those of other angiotensin II receptor blockers. These effects include decreased fasting glucose, glycosylated hemoglobin, interleukin-6, and tumor necrosis factor-α levels. However, no systemic analysis of telmisartan's effects on body fat distribution has been performed. We performed a meta-analysis of randomized controlled telmisartan trials to investigate its effects on body weight, fat distribution, and visceral adipose reduction. A literature search was performed using Embase, MEDLINE, and the Cochrane Library between January 1966 and November 2013. Randomized controlled trials in English and meeting the following criterion were included: random assignment of hypertensive participants with overweight/obesity, metabolic syndrome, or glucose intolerance to telmisartan or control therapy group. Of 651 potentially relevant reports, 15 satisfied the inclusion criterion. While visceral fat area was significantly lower in the telmisartan group than in the control group (weighted mean difference = -18.13 cm(2), 95% C.I. = -27.16 to -9.11, Pχ(2) = 0.19, I(2) = 41%), subcutaneous fat area was similar (weighted mean difference =2.94 cm(2), 95% C.I. = -13.01 to 18.89, Pχ(2) = 0.30, I(2) = 17%). Total cholesterol levels were significantly different between the groups (standardized mean difference = -0.24, 95% C.I. = -0.45 to -0.03, Pχ(2) = 0.0002, I(2) = 67%). Limitations include: (1) limited number of studies, especially those evaluating fat distribution; (2) different imaging modalities to assess visceral fat area (V.F.A.) and subcutaneous fat area (S.F.A.); (3) observed heterogeneity. The findings suggest that telmisartan affected fat distribution, inducing visceral fat reduction, and thus could be useful in hypertensive patients with obesity/overweight, metabolic syndrome, or glucose intolerance.


Histamine-induced fluid efflux in the canine forelimb as affected by carotid occlusion or hemorrhage.
Infusion of catecholamines concurrently with histamine into the forelimb of the dog prevents histamine-induced increases in lymph flow, protein concentration and forelimb weight. This study tested whether the sympathoadrenal discharge of catecholamines induced by carotid occlusion or hemorrhage would similarly prevent histamine's actions in the canine forelimb. Carotid occlusion or hemorrhage, begun after 30 minutes of histamine infusion (4 micrograms base/min), resulted in a reduction in lymph flow but lymph protein concentration was not changed. Infusion of histamine into the forelimb for 30 minutes in control studies caused the forelimb to gain weight. In experimental studies hemorrhage, begun at one minute after the start of histamine significantly reduced this weight gain but occlusion of the carotid arteries at one minute after the start of histamine did not significantly affect weight gain. These results show that sympathetic nerve activity can lower elevated lymph flow and that sympathetic activity due to hemorrhage reduces the accumulation of interstitial fluid caused by histamine. It seems probable that endogenous catecholamines act directly on filtering vessels to lessen histamine-induced increases in permeability. However, a constriction of lymphatic vessels could have contributed to the reduction in lymph flow and we have no evidence against this possibility.


Long-term risedronate treatment normalizes mineralization and continues to preserve trabecular architecture: sequential triple biopsy studies with micro-computed tomography.
The objective of the study was to assess the time course of changes in bone mineralization and architecture using sequential triple biopsies from women with postmenopausal osteoporosis (PMO) who received long-term treatment with risedronate. Transiliac biopsies were obtained from the same subjects (n = 7) at baseline and after 3 and 5 years of treatment with 5 mg daily risedronate. Mineralization was measured using 3-dimensional (3D) micro-computed tomography (CT) with synchrotron radiation and was compared to levels in healthy premenopausal women (n = 12). Compared to the untreated PMO women at baseline, the premenopausal women had higher average mineralization (Avg-MIN) and peak mineralization (Peak-MIN) by 5.8% (P = 0.003) and 8.0% (P = 0.003), respectively, and lower ratio of low to high-mineralized bone volume (BMR-V) and surface area (BMR-S) by 73.3% (P = 0.005) and 61.7% (P = 0.003), respectively. Relative to baseline, 3 years of risedronate treatment significantly increased Avg-MIN (4.9 +/- 1.1%, P = 0.016) and Peak-MIN (6.2 +/- 1.5%, P = 0.016), and significantly decreased BMR-V (-68.4 +/- 7.3%, P = 0.016) and BMR-S (-50.2 +/- 5.7%, P = 0.016) in the PMO women. The changes were maintained at the same level when treatment was continued up to 5 years. These results are consistent with the significant reduction of turnover observed after 3 years of treatment and which was similarly maintained through 5 years of treatment. Risedronate restored the degree of mineralization and the ratios of low- to high-mineralized bone to premenopausal levels after 3 years of treatment, suggesting that treatment reduced bone turnover in PMO women to healthy premenopausal levels. Conventional micro-CT analysis further demonstrated that bone volume (BV/TV) and trabecular architecture did not change from baseline up to 5 years of treatment, suggesting that risedronate provided long-term preservation of trabecular architecture in the PMO women. Overall, risedronate provided sustained benefits on mineralization and architecture, two key determinants of bone strength, over 5 years lending support for its long-term efficacy in fracture risk reduction.


Cluster-Based Multinomial Logistic Regression Model for Health-Related Quality of Life Among People Living with HIV in Brazil.
Improvement in treatment options has increased the survival of people living with HIV (PLHIV). Thus, we evaluated the factors associated with better health-related quality of life (HRQoL) among PLHIV in Brazil. This was a cross-sectional study carried out among 349 PLHIV. Data were collected using an interview-based questionnaire, and HRQoL was assessed by the Brazilian version of the WHOQOL HIV BREF instrument. We used non-hierarchical cluster analysis (K-means) to compile the WHOQOL HIV BREF's overall and domain scores into a unique more multidimensional measure for HRQoL consisting of three clusters: poor, fair and good; associations with clusters of better HRQoL were assessed using multinomial logistic regression models. The mean and median overall HRQoL scores were 15.13 (SD = 3.39) and 16, respectively. The reliability and validity of the Brazilian version of the WHOQOL HIV BREF instrument was confirmed among PLHIV in a non-metropolitan, medium-sized municipality of Brazil, which reaffirmed the cross-cultural validity of this instrument. The factors male sex; heterosexual and asexual orientations; higher individual income; undetectable viral load; absence of any comorbidity and presence of an infectious or a chronic comorbidity, with mental illness as the reference; and never having consumed illegal substances were independently associated with good HRQoL. Thus, the compilation of the WHOQOL HIV BREF's overall and domain scores into a unique multidimensional measure for HRQoL, which this study proposed for the first time, may facilitate more robust interpretations and models of predictors. These differentials could simplify HRQoL as an indicator of health and wellbeing to be routinely used as a key outcome in the clinical management of patients and in the global monitoring of health system responses to HIV.


Biomarkers in Chronic Obstructive Pulmonary Disease: Emerging Roles of Eosinophils and Procalcitonin.
Antibiotics can improve the prognosis in patients with exacerbation of chronic obstructive pulmonary disease. However, the overuse of antibiotics can carry serious adverse effects for patients (gastrointestinal infections) and for society (bacterial resistance). Likewise, systemic corticosteroids may also help these patients, but also carries severe adverse effects like osteoporosis, muscle loss, and diabetes, in many patients. Whenever safe methods exist to reduce these two treatment modalities, they should be implemented. The blood biomarkers procalcitonin and the fraction of leukocytes known as eosinophil granulocytes have been proven in randomized controlled trials (RCTs), to effectively, significantly, and substantially assist in reducing the use of these two potent, yet toxic medication types. In this review, the background and main clinical results are discussed, explaining the rationale for biomarker-guided clinical decisions. Also, the main expected effects, their sizes, and importantly the limitations to such a strategy are described. Clinical evidence is prioritized with main weight on RCTs and meta-analyses of these and regarding outcomes, and focus is set on the safety of such a biomarker-guided strategy, as well as the effects on medicine reduction. In an epoch of increasing demands to physicians from patients and politicians to cure and reduce symptoms, the Hippocratic phrase of "primum non nocere" or "first, do no harm" seems more than ever of contemporary importance.