Ten years of marketing approvals of anticancer drugs in Europe: regulatory policy and guidance documents need to find a balance between different pressures.
Despite important progress in understanding the molecular factors underlying the development of cancer and the improvement in response rates with new drugs, long-term survival is still disappointing for most common solid tumours. This might be because very little of the modest gain for patients is the result of the new compounds discovered and marketed recently. An assessment of the regulatory agencies' performance may suggest improvements. The present analysis summarizes and evaluates the type of studies and end points used by the EMEA to approve new anticancer drugs, and discusses the application of current regulations. This report is based on the information available on the EMEA web site. We identified current regulatory requirements for anticancer drugs promulgated by the agency and retrieved them in the relevant directory; information about empirical evidence supporting the approval of drugs for solid cancers through the centralised procedure were retrieved from the European Public Assessment Report (EPAR). We surveyed documents for drug applications and later extensions from January 1995, when EMEA was set up, to December 2004. We identified 14 anticancer drugs for 27 different indications (14 new applications and 13 extensions). Overall, 48 clinical studies were used as the basis for approval; randomised comparative (clinical) trial (RCT) and Response Rate were the study design and end points most frequently adopted (respectively, 25 out of 48 and 30 out of 48). In 13 cases, the EPAR explicitly reported differences between arms in terms of survival: the range was 0-3.7 months, and the mean and median differences were 1.5 and 1.2 months. The majority of studies (13 out of 27, 48%) involved the evaluation of complete and/or partial tumour responses, with regard to the end points supporting the 27 indications. Despite the recommendations of the current EMEA guidance documents, new anticancer agents are still often approved on the basis of small single arm trials that do not allow any assessment of an 'acceptable and extensively documented toxicity profile' and of end points such as response rate, time to progression or progression-free survival which at best can be considered indicators of anticancer activity and are not 'justified surrogate markers for clinical benefit'. Anticipating an earlier than ideal point along the drug approval path and the use of not fully validated surrogate end points in nonrandomised trials looks like a dangerous shortcut that might jeopardise consumers' health, leading to unsafe and ineffective drugs being marketed and prescribed. The present Note for Guidance for new anticancer agents needs revising. Drugs must be rapidly released for patients who need them but not be at the expense of adequate knowledge about the real benefit of the drugs.


A detailed molecular network map and model of the NLRP3 inflammasome.
The NLRP3 inflammasome is a key regulator of inflammation that responds to a broad range of stimuli. The exact mechanism of activation has not been determined, but there is a consensus on cellular potassium efflux as a major common denominator. Once NLRP3 is activated, it forms high-order complexes together with NEK7 that trigger aggregation of ASC into specks. Typically, there is only one speck per cell, consistent with the proposal that specks form - or end up at - the centrosome. ASC polymerisation in turn triggers caspase-1 activation, leading to maturation and release of IL-1β and pyroptosis, i.e., highly inflammatory cell death. Several gain-of-function mutations in the NLRP3 inflammasome have been suggested to induce spontaneous activation of NLRP3 and hence contribute to development and disease severity in numerous autoinflammatory and autoimmune diseases. Consequently, the NLRP3 inflammasome is of significant clinical interest, and recent attention has drastically improved our insight in the range of involved triggers and mechanisms of signal transduction. However, despite recent progress in knowledge, a clear and comprehensive overview of how these mechanisms interplay to shape the system level function is missing from the literature. Here, we provide such an overview as a resource to researchers working in or entering the field, as well as a computational model that allows for evaluating and explaining the function of the NLRP3 inflammasome system from the current molecular knowledge. We present a detailed reconstruction of the molecular network surrounding the NLRP3 inflammasome, which account for each specific reaction and the known regulatory constraints on each event as well as the mechanisms of drug action and impact of genetics when known. Furthermore, an executable model from this network reconstruction is generated with the aim to be used to explain NLRP3 activation from priming and activation to the maturation and release of IL-1β and IL-18. Finally, we test this detailed mechanistic model against data on the effect of different modes of inhibition of NLRP3 assembly. While the exact mechanisms of NLRP3 activation remains elusive, the literature indicates that the different stimuli converge on a single activation mechanism that is additionally controlled by distinct (positive or negative) priming and licensing events through covalent modifications of the NLRP3 molecule. Taken together, we present a compilation of the literature knowledge on the molecular mechanisms on NLRP3 activation, a detailed mechanistic model of NLRP3 activation, and explore the convergence of diverse NLRP3 activation stimuli into a single input mechanism.


Alcohol abuse and HIV infection: role of DRD2.
According to a survey from the HIV Cost and Services Utilization Study (HCSUS), approximately 53% of HIV-infected patients reported drinking alcohol and 8% were classified as heavy drinkers. The role of alcohol as a risk factor for HIV infection has been widely studied and recent research has found a significant association between heavy alcohol consumption and lower levels of CD4 T cells among HIV-infected alcoholics. Although there is evidence on the role of alcohol as a risk factor for HIV transmission and disease progression, there is a need for population studies to determine the genetic mechanisms that affect alcohol's role in HIV disease progression. One of the mechanisms of interest is the dopaminergic system. To date, the effects of dopamine on HIV neuroimmune pathogenesis are not well understood; however, dopaminergic neural degeneration due to HIV is known to occur by viral invasion into the brain via immune cells, and modulation of dopamine in the CNS may be a common mechanism by which different types of substances of abuse impact HIV disease progression. Although previous studies have shown an association of D(2) dopamine receptor (DRD2) polymorphisms with severity of alcohol dependence, the expression of this allele risk on HIV patients with alcohol dependence has not been systematically explored. In the current study, DRD2 Taq1A and C957T SNP genotyping analyses were performed in 165 HIV-infected alcohol abusers and the results were examined with immune status and CD4 counts.


Nutritional improvement of soy flour.
Heating soy flour at 45 degrees C in pH 8.5 Tris buffer for 1 hour in the presence of cysteine or N-acetylcysteine followed by dialysis to remove unreacted thiols resulted in the introduction of new half-cystine residues and lowered trypsin inhibitor content from 37.5 to 6.8 or 4.8 mg/g, respectively, compared to only 14.8 mg/g in the absence of sulfhydryl compounds. Presence of cysteine or N-acetylcysteine during treatment increased protein efficiency ratio (PER) from 0.95 to 2.01 or 2.20, respectively. Corresponding values at 65 degrees C are 1.61, 2.43 and 2.02; and at 75 degrees C, 2.14, 2.53 and 3.19, respectively. Proteins are modified through formation of mixed disulfide bonds, which leads to loss of inhibitory activity and increased protein digestibility and nutritive value.


Triangle of COVID, anxiety and menstrual cycle.
This study aimed to examine whether the menstrual cycles of women with regular cycles have been affected by the COVID 19 pandemic. This cross-sectional online survey study evaluated the menstrual cycle characteristics of women in the reproductive phase of their lives during the COVID-19 pandemic in May 2020. Changes before and after COVID-19 were evaluated with a paired sample t-test and chi-square test. A Pearson correlation test was used to show the relationships between variables. The study was completed with 263 participants in total. The average age of the participants was 26.3 ± 6.9 (18-45). Participants' duration of period (p = .003) and pads used per day (p = .002) decreased compared to their experience before the COVID-19 outbreak. The mean total MSQ score was 3.1 ± 0.8 (0-4), mean STAI-1 score was 45.1 ± 9 (20-78) and mean STAI-II score was 43.3 ± 5.9 (30-69). It was found that STAI-I and STAI-II scores showed a positive correlation with MSQ-Somatic subgroup scores(r = 0.153, p = .013; 0.190, p = .002) and MSQ-Total scores (0.144, p = .020; 0.175, p = .004). With the COVID-19 pandemic, increased anxiety scores increased women's menstrual symptoms while the length of periods and the number of pads used decreased.Impact statementWhat is already known on this subject? The COVID 19 pandemic, has caused enormous psychological distress potentially resulting in the development of posttraumatic stress disorder. The menstrual cycle is a process that can be affected by psychological stress.What do the results of this study add? This is the first study to examine the relationship between stress caused by the COVID-19 pandemic and the menstrual cycle. The increases in the degree of anxiety and stress as a result of the COVID-19 outbreak was found to be high enough to affect the characteristics of the menstrual cycle in the women surveyed.What are the implications of these findings for clinical practice and/or further research? In a prospective study intended to be carried out after the outbreak ends in the future, it will be possible to evaluate whether the menstrual cycle parameters return to their former order and consequently this hypothesis will be able to be more definitively confirmed.


Self-perception of body weight among high school students in Taipei, Taiwan.
Self-perception of body weight and other weight-related factors were assessed among 2665 Taipei, Taiwan high school students. A high percent of the girls (70.7%) and boys (42.2%) reported that they were too fat and these percentages were much higher than those reported by U.S. students in a recent Youth Risk Behavior Survey. In addition, only 13.2% of girls and 22.0% of boys reported being completely satisfied with their weight and the level of dissatisfaction with weight appeared to be greater than among U.S. students. Yet, in comparison to U.S. students, the Taiwanese students were considerably less likely than their U.S. counterparts to engage in weight management practices (e.g., dieting, eating less food, using diet pills). Taiwanese students with a self-perception of being too fat were more likely than those with perceptions of being just right or too thin to engage in weight management practices, to be dissatisfied with their weight, feel that they were unattractive, estimate that their same-sex peers were trying to lose weight, and have a higher body mass index. The findings from this study showed a relationship between self-perception of body size and engaging in weight control behaviors was consistent with other research. It suggested that self-perception of body weight, more so than objective weight status, was predictive of weight loss behavior and also negative psychological outcomes associated with poor body weight image. As a result, self-perception of weight may be an important point of focus for the design and implementation of clinical and public health initiatives targeted at this adolescent population as well as others.


Reduced growth, increased vascular area, and reduced response to cisplatin in CD13-overexpressing human ovarian cancer xenografts.
Expression of aminopeptidase N/CD13 can be detected in several solid tumor types. Thus far, the role of CD13 in ovarian cancer has not been studied. We have investigated the expression pattern and biological function of CD13 in ovarian cancer. First, we studied the expression of CD13 in ovarian cancer tissue of 15 patients representing three different histological types (5 patients each) by immunohistochemistry. We then stably transfected the IGROV-1 human ovarian cancer cell line with a CD13 expression vector and examined the biological effect of CD13 in vitro and in vivo. The expression of CD13 in ovarian cancer was associated with the histological subtype: CD13 expression in tumor cells was observed in 80-100% of the patients with a serous or mucinous carcinoma and in only 20% of the clear cell carcinoma patients. In all patients' tumor samples, CD13-positive blood vessels were present. CD13 overexpression in IGROV-1 cells did not affect in vitro cell growth and sensitivity to doxorubicin, cisplatin, or gemcitabine. CD13 overexpression reduced invasion in Matrigel, which appeared to be independent of the aminopeptidase activity of CD13. Furthermore, the growth rate of IGROV-1/CD13 xenografts was reduced. The area of the vessel lumens was enlarged in a small percentage of vessels in the CD13-overexpressing xenografts. In addition, the CD13-overexpressing tumors were less sensitive to cisplatin. CD13 is expressed in tumor as well as endothelial cells in human ovarian cancer. Our results suggest that CD13 overexpression affects ovarian cancer growth, vascular architecture, and response to chemotherapy. Further elucidation of the mechanism of the observed effects of CD13 is warranted to better understand its role in the pathophysiology of ovarian cancer.


Overview of Phlorotannins' Constituents in Fucales.
Fucales are an order within the Phaeophyceae that include most of the common littoral seaweeds in temperate and subtropical coastal regions. Many species of this order have long been a part of human culture with applications as food, feedand remedies in folk medicine. Apart from their high nutritional value, these seaweeds are also a well-known reservoir of multiple bioactive compounds with great industrial interest. Among them, phlorotannins, a unique and diverse class of brown algae-exclusive phenolics, have gathered much attention during the last few years due to their numerous potential health benefits. However, due to their complex structural features, combined with the scarcity of standards, it poses a great challenge to the identification and characterization of these compounds, at least with the technology currently available. Nevertheless, much effort has been taken towards the elucidation of the structural features of phlorotannins, which have resulted in relevant insights into the chemistry of these compounds. In this context, this review addresses the major contributions and technological advances in the field of phlorotannins extraction and characterization, with a particular focus on Fucales.


Detection of extended-spectrum beta-lactamases by using boronic acid as an AmpC beta-lactamase inhibitor in clinical isolates of Klebsiella spp. and Escherichia coli.
We evaluated highly sensitive methods using boronic acid (BA) to detect extended-spectrum beta-lactamase (ESBL) production. A total of 182 clinical isolates of Klebsiella spp. (n = 118) and Escherichia coli (n = 64) were analyzed: 62 harbored only ESBLs, 80 harbored both ESBLs and plasmid-mediated AmpC beta-lactamases (pAmpCs), and 40 harbored only pAmpCs. The CLSI confirmatory test detected all isolates that produce only ESBLs but detected 85% of isolates that produce both enzymes. When a >/=5-mm increase in the zone diameter of either the cefotaxime (CTX) or the ceftazidime (CAZ) disk in the presence of both clavulanic acid (CA) and BA was considered to be a positive result, the test detected all isolates that harbor ESBLs (+/- pAmpCs) but showed frequent false-positive results (50%) for isolates that produce only pAmpCs. Meanwhile, when a >/=3-mm increase in the zone diameter of either the CTX/BA or the CAZ/BA disk in the presence of CA was considered to be a positive result, the test also detected all isolates that harbor ESBLs (+/- pAmpCs) and showed less frequent false-positive results (5%) in isolates that produce only pAmpCs. The latter new interpretive guideline has enhanced detection of ESBLs in clinical isolates of Klebsiella spp. and Escherichia coli and allowed detection of an ESBL even when potentially masked by a pAmpC.


Genetic ablation of calcium-independent phospholipase A(2)beta causes hypercontractility and markedly attenuates endothelium-dependent relaxation to acetylcholine.
Activation of phospholipases leads to the release of arachidonic acid and lysophospholipids that play prominent roles in regulating vasomotor tone. To identify the role of calcium-independent phospholipase A(2)beta (iPLA(2)beta) in vasomotor function, we measured vascular responses to phenylephrine (PE) and ACh in mesenteric arterioles from wild-type (WT; iPLA(2)beta(+/+)) mice and those lacking the beta-isoform (iPLA(2)beta(-/-)) both ex vivo and in vivo. Vessels isolated from iPLA(2)beta(-/-) mice demonstrated increased constriction to PE, despite lower basal smooth muscle calcium levels, and decreased vasodilation to ACh compared with iPLA(2)beta(+/+) mice. PE constriction resulted in initial intracellular calcium release with subsequent steady-state constriction that depended on extracellular calcium influx. Endothelial denudation had no effect on vessel tone or PE-induced constriction although the dilation to ACh was significantly reduced in iPLA(2)beta(+/+) vessels. In contrast, vessels from iPLA(2)beta(-/-) constricted by 54% after denudation, indicating smooth muscle hypercontractility. In vivo, blood pressure, resting vessel diameter, and constriction of mesenteric vessels to PE were not different in iPLA(2)beta(-/-) vessels compared with WT mouse vessels. However, relaxation after ACh administration in situ was attenuated, indicating an endothelial inability to induce dilation in response to ACh. In cultured endothelial cells, inhibition of iPLA(2)beta with (S)-(E)-6-(bromomethylene)tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one (BEL) decreased endothelial nitric oxide synthase phosphorylation and reduced endothelial agonist-induced intracellular calcium release as well as extracellular calcium influx. We conclude that iPLA(2)beta is an important mediator of vascular relaxation and intracellular calcium homeostasis in both smooth muscle and endothelial cells and that ablation of iPLA(2)beta causes agonist-induced smooth muscle hypercontractility and reduced agonist-induced endothelial dilation.


Are fear-avoidance beliefs related to the inception of an episode of back pain? A prospective study.
Abstract Fear-avoidance beliefs and catastrophizing have been implicated in chronic pain and theoretical models have been developed that feature these factor in the transition from acute to chronic pain. However, little has been done to determine whether these factors occur in the general population or whether they arc associated with the inception of an episode of neck or back pain. The aim of this study was to evaluate prospectively the effects of fear-avoidance beliefs and catastrophizing on the development of an episode of self-reported pain and associated physical functioning. To achieve this, we selected a sample of 415 people from the general population who reported no spinal pain during the past year. At the pretest a battery of questionnaires was administered to assess beliefs about pain and activity and it featured the Pain Catastrophizing Scale and a modified version of the Fear-Avoidance Beliefs Questionnaire. One year later outcome was evaluated by self-reports of the occurrence of a pain episode as well as a self-administered physical function test. The results showed that scores on both fear-avoidme and cabstrophizing were quite low. During the one year follow-up, 19% of the sample suffered an episode of back pain. Those with scores above the median on fear-avoidance beliefs at the pretest had twice the risk of suffering an episode of back pain and a 1.7 times higher risk of lowered physical function at the follow-up. Catastrophizing was somewhat less salient, increasing the risk of pain or lowered function by 1.5. but with confidence intervals falling below unity. These data indicate that fear-avoidance beliefs may be involved at a very early pint in the development of pain and associated activity problems in people with back pain. Theoretically. our results support the idea that fear-avoidance beliefs may develop in an interaction with the experience of pain. Clinically, the results suggest that catastrophizing and particularly fear-avoidance beliefs are important in the development of a pain problem and might be of use in screening procedures.


Berberis Fruit Extract and Biochemical Parameters in Patients With Type II Diabetes.
Diabetes mellitus is a common medical problem. There is in fact a growing body of literature on plants used for the treatment of diabetes. Plant materials attracted considerable interest of scientists. In this respect, in the past few years, attempts were made to use natural plant products for the treatment of patients with diabetes. The aim of this study was to investigate the effect of Berberis fruit extract to achieve glycemic control in patients with Type II diabetes. This study was performed between July 2010 and April 2013. Thirty patients of type II diabetes admitted to Ayatollah Rohhani hospital were recruited. Patients' sera were collected for the assessment of glucose and HbA1c values. Biochemical analyses were performed before and after treatment by Berberis fruit extract. Biochemical parameters were measured by spectrophotometric method (Jenway uv/vis, 6505 model, Dunmow, UK). Glucose level was measured by glucose oxidase method kit (Pars Azmoon, Tehran, IR Iran). Serum total cholesterol and triglycerides were measured using standard biochemical kits (Pars Azmoon, Tehran, Iran). Blood glycated hemoglobin level was measured by using Elisa kit (Bioassay technology laboratory, Elisa kit). Experiments were performed in triplicate in at least three separate experiments. Our findings demonstrated that patients with type 2 diabetes who received barberry fruit had significant reduction in serum glucose to 136.15 ± 32.8 mg/dL and decreased HbA1c levels to 7.07 ± 1.21 mg/dL, during the 8 weeks of study. This investigation revealed that Berberis fruit extract has beneficial metabolic effects in patients with type II diabetes. Barberry may improve glucose catabolism via glycolysis pathway, stimulate insulin secretion or improve insulin function and finally decrease glucose uptake. Our results indicated that Berberis fruit regulates glucose metabolism in patients with type 2 diabetes.


MicroRNA-221-3p promotes post-burn HUVEC proliferation, migration, and angiogenesis by regulating CDKN1B.
Previous studies have shown that miR-221-3p plays an important role in vascular remodeling, but it is unclear whether it contributes to angiogenesis after burn injury. The purpose of this study was to investigate the effect of miR-221-3p on angiogenesis in HUVECs after burn injury and to reveal its underlying molecular mechanism. The burn HUVECs model was established by heat treatment. Plasmid or oligonucleotide transfection altered the expression of miR-221-3p and CDKN1B in HUVECs. MTT, colony formation, Transwell, flow cytometry, and tube formation experiments were applied to assess the proliferation, migration, apoptosis, cell cycle, and tube formation capacity of HUVECs. miR-221-3p, CDKN1B, Ki-67, and PCNA expression was assessed by RT-qPCR or Western blot. The dual-luciferase reporter assay verified the targeting relationship between miR-221-3p and CDKN1B. miR-221-3p was lowly expressed and CDKN1B was highly expressed in burn HUVECs. Overexpression of miR-221-3p promoted the proliferation, migration, and tube formation ability of burn HUVECs and inhibited apoptosis and the proportion of cells in the G0/G1 phase, whereas overexpression of CDKN1B had the opposite effect. Knockdown of miR-221-3p further inhibited the angiogenic capacity of burn HUVECs, but this effect was reversed by knockdown of CDKN1B. Mechanistically, miR-221-3p targeted CDKN1B. miR-221-3p improves the angiogenesis of burn HUVECs by targeting CDKN1B expression, and the miR-221-3p/CDKN1B axis may serve as a potential molecular target for future burn therapy.


A comparison of various Haemophilus somnus strains.
Sixty-eight Haemophilus somnus strains isolated from the bovine in Canada and the U.S.A. were compared. In media enriched with 5% ovine serum, 5% bovine serum and 10% yeast extract, H. somnus fermented glucose, levulose, maltose, mannitol, mannose, sorbitol, trehalose and xylose, but failed to ferment arabinose, dulcitol, galactose, inositol, lactose, raffinose, rhamnose, salicin and sucrose. The organisms acidified litmus milk, produced cytochrome oxidase, indole and hydrogen sulfide (H(2)S) and reduced nitrates to nitrites. The motility, methyl-red, acetylmethyl-carbinol urease catalase, citrate, malonate, lysine, ornithine and arginine tests were negative. Haemophilus somnus was resistant to lincomycin, neomycin and triple sulfa, but susceptible to ampicillin, chloramphenicol, streptomycin, penicillin and tetracycline. No antigenic differences were noted between strains when tested against rabbit antisera of eight strains using agglutination, complement-fixation, immunodiffusion and counterimmunoelectrophoresis tests. Low titre cross-reactions were found in the agglutination tests with some of the anti-H. somnus rabbit sera with Actinobacillus lignieresi and Moraxella bovis. No distinct antigenic similarities to nine other species of pathogenic bacteria of animal origin were found. No difference was observed between H. somnus isolates from Ontario and those from western Canada and the U.S.A.


High-throughput sequencing identifies STAT3 as the DNA-associated factor for p53-NF-kappaB-complex-dependent gene expression in human heart failure.
Genome-wide maps of DNA regulatory elements and their interaction with transcription factors may form a framework for understanding regulatory circuits and gene expression control in human disease, but how these networks, comprising transcription factors and DNA-binding proteins, form complexes, interact with DNA and modulate gene expression remains largely unknown. Using microRNA-21 (mir-21), which is an example of genes that are regulated in heart failure, we performed chromatin immunoprecipitation (ChIP) assays to determine the occupancy of transcription factors at this genetic locus. Tissue ChIP was further performed using human hearts and genome-wide occupancies of these transcription factors were analyzed by high-throughput sequencing. We show that the transcription factor p53 piggy-backs onto NF-kappaB/RELA and utilizes the kappaB-motif at a cis-regulatory region to control mir-21 expression. p53 behaves as a co-factor in this complex because despite a mutation in its DNA binding domain, mutant p53 was still capable of binding RELA and the cis-element, and inducing mir-21 expression. In dilated human hearts where mir-21 upregulation was previously demonstrated, the p53-RELA complex was also associated with this cis-element. Using high-throughput sequencing, we analyzed genome-wide binding sites for the p53-RELA complex in diseased and control human hearts and found a significant overrepresentation of the STAT3 motif. We further determined that STAT3 was necessary for the p53-RELA complex to associate with this cis-element and for mir-21 expression. Our results uncover a mechanism by which transcription factors cooperate in a multi-molecular complex at a cis-regulatory element to control gene expression.


Long-term effects of exercise on the range of motion of aging women.
Loss of mobility is important to everyone, particularly the elderly. The purpose of this retrospective, longitudinal study was to determine the long-term (5-year) effects of exercise on the shoulder and hip range of motion of a group of 12 women aged 50-71 years. Shoulder flexion, shoulder extension, shoulder transverse extension, hip flexion, and hip rotation were measured a minimum of five times throughout the 5-year period. A flexometer was used to measure range of motion. The results of the study indicated that range of motion increased significantly (p < 0.05) throughout the study in four of the five measurements. The fifth measurement (shoulder flexion) also improved but not significantly. These data indicate that aging women can improve and/or maintain shoulder and hip range of motion through participation in regular exercise done thee times per week for 5 years. J Orthop Sports Phys Ther 1992;16(1):37-42.


Model system for high-throughput screening of novel human immunodeficiency virus protease inhibitors in Escherichia coli.
Novel human immunodeficiency virus (HIV) protease inhibitors are urgently needed for combating the drug-resistance problem in the fight against AIDS. To facilitate lead discovery of HIV protease inhibitors, we have developed a safe, convenient, and cost-effective Escherichia coli-based assay system. This E. coli-based system involves coexpression of an engineered beta-galactosidase as an HIV protease substrate and the HIV protease precursor comprising the transframe region and the protease domain. Autoprocessing of the HIV protease precursor releases the mature HIV protease. Subsequently, the HIV protease cleaves beta-galactosidase, resulting in a loss of the beta-galactosidase activity, which can be detected in high-throughput screens. Using Food and Drug Administration-approved HIV protease inhibitors, this E. coli-based system is validated as a surrogate screening system for identifying inhibitors that not only possess inhibitory activity against HIV protease but also have solubility and permeability for in vivo activity. The usefulness of the E. coli-based system was demonstrated with the identification of a novel HIV protease inhibitor from a library of compounds that were prepared by an amide-forming reaction with transition-state analog cores. A novel inhibitor with a sulfonamide core of amprenavir, E2, has shown good correlation with the in vitro enzymatic assay and in vivo E. coli-based system. This system can also be used to generate drug resistance profiles that could be used to suggest therapeutic uses of HIV protease inhibitors to treat the drug-resistant HIV strains. This simple yet efficient E. coli system not only represents a screening platform for high-throughput identification of leads targeting the HIV proteases but also can be adapted to all other classes of proteases.


Delayed contrast-enhanced MRI of the coronary artery wall in takayasu arteritis.
Takayasu arteritis (TA) is a rare form of chronic inflammatory granulomatous arteritis of the aorta and its major branches. Late gadolinium enhancement (LGE) with magnetic resonance imaging (MRI) has demonstrated its value for the detection of vessel wall alterations in TA. The aim of this study was to assess LGE of the coronary artery wall in patients with TA compared to patients with stable CAD. We enrolled 9 patients (8 female, average age 46±13 years) with proven TA. In the CAD group 9 patients participated (8 male, average age 65±10 years). Studies were performed on a commercial 3T whole-body MR imaging system (Achieva; Philips, Best, The Netherlands) using a 3D inversion prepared navigator gated spoiled gradient-echo sequence, which was repeated 34-45 minutes after low-dose gadolinium administration. No coronary vessel wall enhancement was observed prior to contrast in either group. Post contrast, coronary LGE on IR scans was detected in 28 of 50 segments (56%) seen on T2-Prep scans in TA and in 25 of 57 segments (44%) in CAD patients. LGE quantitative assessment of coronary artery vessel wall CNR post contrast revealed no significant differences between the two groups (CNR in TA: 6.0±2.4 and 7.3±2.5 in CAD; p = 0.474). Our findings suggest that LGE of the coronary artery wall seems to be common in patients with TA and similarly pronounced as in CAD patients. The observed coronary LGE seems to be rather unspecific, and differentiation between coronary vessel wall fibrosis and inflammation still remains unclear.


Cost-Effectiveness of Sarilumab Added to Methotrexate in the Treatment of Adult Patients with Moderately to Severely Active Rheumatoid Arthritis Who Have Inadequate Response or Intolerance to Tumor Necrosis Factor Inhibitors.
Despite a substantial number of treatment options in rheumatoid arthritis (RA) following tumor necrosis factor inhibitor (TNFi) inadequate response or intolerance (TNF-IR), a lack of clarity on the optimal approach remains. Sarilumab, a human monoclonal anti-interleukin-6 receptor alpha antibody, can be used as monotherapy or in combination with methotrexate or other conventional synthetic disease-modifying anti-rheumatic drugs (DMARDs) in TNF-IR patients. To conduct a cost-utility analysis from a U.S. health care system perspective for sarilumab subcutaneous 200 mg + methotrexate versus abatacept + methotrexate or a bundle of TNFi + methotrexate for treatment of adult patients with moderately to severely active RA and TNF-IR. Analysis was conducted via individual patient simulation based on patient profiles from the TARGET trial (NCT01709578); a 6-month decision tree was followed by lifetime semi-Markov model with 6-month cycles. Treatment response at 6 months, informed by network meta-analysis, was based on American College of Rheumatology (ACR) 20/50/70 criteria; patients achieving ≥ ACR20 continued with current therapy, and other patients moved to the next line of biologic DMARD therapy or conventional synthetic DMARD palliative treatment. Direct costs included wholesale acquisition drug costs and administration and routine care costs. Routine care costs and quality-adjusted life-years (QALYs) were estimated by predicting the Health Assessment Questionnaire Disability Index score based on treatment response and were imputed from published equations. Sarilumab + methotrexate dominated the TNFi bundle + methotrexate, achieving lower costs ($319,324 vs. $356,096) and greater effectiveness (4.27 vs. 4.15 QALYs), and was on the cost-efficiency frontier with abatacept + methotrexate ($360,211 and 4.29 QALYs). Abatacept + methotrexate was not cost-effective versus sarilumab + methotrexate. Scenario analyses indicated the results were robust; sarilumab + methotrexate became dominant against abatacept + methotrexate after reduced model horizon, minimum response based on ACR50 or ACR70, or time to discontinuation per treatment class. Sarilumab + methotrexate was also dominant versus the TNFi bundle; when class-specific time to treatment discontinuation was specified, sarilumab remained cost-effective with an incremental cost-effectiveness ratio of $36,894. Sarilumab + methotrexate can be considered an economically dominant (more effective, less costly) option versus a second TNFi + methotrexate; compared with abatacept + methotrexate, it is a less costly but less effective option for patients with moderately to severely active RA who have previously failed TNFi. This study was funded by Sanofi and Regeneron Pharmaceuticals. Kiss and Gal are employees of Evidera, which received consulting fees from Sanofi/Regeneron for conducting this study. Muszbek was employed by Evidera at the time of this study. Kuznik and Chen are current employees of and stockholders in Regeneron Pharmaceuticals. Fournier is an employee of and stockholder in Sanofi. Proudfoot is a former employee of and current stockholder in Sanofi and current employee and stockholder in ViiV Healthcare/GlaxoSmithKline. Michaud has received grant funding from Pfizer and the Rheumatology Research Foundation. The sponsors were involved in the study design, collection, analysis, and interpretation of data as well as data checking of information provided in the manuscript. The authors had unrestricted access to study data, were responsible for all content and editorial decisions, and received no honoraria related to the development of this publication.


Single embryo transfer (set) not only leads to a reduction in twinning rates after IVF/ICSI, but also improves obstetrical and perinatal outcome of singletons.
This work consists of two parts. In the first part we investigated the influence of first trimester bleeding on perinatal outcome after assisted reproductive technologies (ART) and we found that first-trimester bleeding not necessarily leads to spontaneous abortion, as in spontaneous pregnancies. On the other hand, first-trimester bleeding clearly is associated with an impaired pregnancy outcome, and as an independent risk factor for obstetrical and perinatal outcome, it should be taken into account in studies analysing pregnancy outcome after ART. The fact that the incidence of first trimester bleeding increases with the number of embryos transferred is another argument to move to single embryo transfer (SET). Next to a very effective reduction in multiple pregnancy rates, SET indeed seems to be beneficial in terms of pregnancy outcome characteristics in singleton pregnancies as well. In the second part of this work we indeed have demonstrated that mean birthweight is significantly higher in SET singletons compared with DET singletons. Also the incidence of preterm birth and low birthweight is clearly reduced when SET is compared with DET. A possible explanation for this outcome can be the 'avoidance' of vanishing twins in IVF/ICSI procedures when SET is applied, but this hypothesis needs further investigation. A more frequent application of SET in assisted reproductive technologies clinics could be an essential step to reduce differences in outcome between spontaneous and assisted reproduction children.


Contralateral muscle activity and fatigue in the human first dorsal interosseous muscle.
During effortful unilateral contractions, muscle activation is not limited to the target muscles but activity is also observed in contralateral muscles. The amount of this associated activity is depressed in a fatigued muscle, even after correction for fatigue-related changes in maximal force. In the present experiments, we aimed to compare fatigue-related changes in associated activity vs. parameters that are used as markers for changes in central nervous system (CNS) excitability. Subjects performed brief maximal voluntary contractions (MVCs) with the index finger in abduction direction before and after fatiguing protocols. We followed changes in MVCs, associated activity, motor-evoked potentials (MEP; transcranial magnetic stimulation), maximal compound muscle potentials (M waves), and superimposed twitches (double pulse) for 20 min after the fatiguing protocols. During the fatiguing protocols, associated activity increased in contralateral muscles, whereas afterwards the associated force was reduced in the fatigued muscle. This force reduction was significantly larger than the decline in MVC. However, associated activity (force and electromyography) remained depressed for only 5-10 min, whereas the MVCs stayed depressed for over 20 min. These decreases were accompanied by a reduction in MEP, MVC electromyography activity, and voluntary activation in the fatigued muscle. According to these latter markers, the decrease in CNS motor excitability lasted much longer than the depression in associated activity. Differential effects of fatigue on (associated) submaximal vs. maximal contractions might contribute to these differences in postfatigue behavior. However, we cannot exclude differences in processes that are specific to either voluntary or to associated contractions.


Neural dysfunction during temporal discounting in paediatric Attention-Deficit/Hyperactivity Disorder and Obsessive-Compulsive Disorder.
Both Attention-Deficit/Hyperactivity Disorder (ADHD) and Obsessive-Compulsive Disorder (OCD) are associated with choice impulsivity, i.e. the tendency to prefer smaller immediate rewards over larger delayed rewards. However, the extent to which this impulsivity is mediated by shared or distinct underlying neural mechanisms is unclear. Twenty-six boys with ADHD, 20 boys with OCD and 20 matched controls (aged 12-18) completed an fMRI version of an individually adjusted temporal discounting (TD) task which requires choosing between a variable amount of money now or £100 in one week, one month or one year. Activations to immediate and delayed reward choices were compared between groups using a three-way ANCOVA. ADHD patients had steeper discounting rates on the task relative to controls. OCD patients did not differ from controls or patients with ADHD. Patients with ADHD and OCD showed predominantly shared activation deficits during TD in fronto-striato-insular-cerebellar regions responsible for self-control and temporal foresight, suggesting that choice impulsivity is mediated by overlapping neural dysfunctions in both disorders. OCD patients alone showed dysfunction relative to controls in right orbitofrontal and rostrolateral prefrontal cortex, extending previous findings of abnormalities in these regions in OCD to the domain of choice impulsiveness.


DOT1L decelerates the development of osteoporosis by inhibiting SRSF1 transcriptional activity via microRNA-181-mediated KAT2B inhibition.
Our study explored the function of DOT1L in osteoporosis (OP) via the microRNA (miR)-181/KAT2B/SRSF1 axis. Osteoclast (OC) number was evaluated via TRAP staining, and serum CTXI, PINP, and ALP contents were tested by ELISA. Following identification of bone marrow mesenchymal stem cells (BMSCs), OC differentiation was induced by M-CSF and RANKL, followed by the detection of OC differentiation and the expression of bone resorption-related genes, DOT1L, miR-181, KAT2B, and SRSF1. Overexpressed DOT1L or miR-181 stimulated calcified nodule formation and increased alkaline phosphatase activity and osteogenic marker gene expression. KAT2B knockdown enhanced the osteogenic differentiation of BMSCs by reducing SRSF1 acetylation. The enhancement of OC differentiation induced by overexpressed SRSF1 was inhibited by simultaneous DOT1L or miR-181 overexpression. DOT1L suppressed OP development in vivo via the miR-181/KAT2B/SRSF1 axis. DOT1L overexpression slowed down bone loss and promoted bone formation via the miR-181/KAT2B/SRSF1 axis, thereby alleviating OP development.


Analysis of LINC01314 and miR-96 Expression in Colorectal Cancer Patients via Tissue Microarray-Based Fluorescence In Situ Hybridization.
A tissue microarray (TMA) containing 76 individual colorectal tumor samples and 28 adjacent normal samples was constructed, and the expression levels of LINC01314 and miR-96 were detected by fluorescence in situ hybridization. The expression levels of both LINC01314 and miR-96 were upregulated in CRC tissues and were associated with vascular metastasis (p < 0.05). A significantly positive correlation was observed between LINC01314 and miR-96 expression in tumor tissues (p < 0.001, r = 0.870). Dominant expression of LINC01314 was a risk factor for both blood vessel invasion (p < 0.05) and poor 5-year survival (p = 0.001, hazard ratio = 4.144). The Kaplan-Meier analysis indicated that patients with LINC01314-dominant expression exhibited worse 5-year survival rates than those with miR-96-dominant expression (p < 0.05). The expression patterns of both LINC01314 and miR-96 may be diagnostic of, and prognostic for, CRC. These findings will facilitate further exploration of the molecular mechanism of lncRNAs in CRC.


Comparative effectiveness and predictors of response to tumour necrosis factor inhibitor therapies in rheumatoid arthritis.
Adalimumab, etanercept and infliximab are effective TNF inhibitors (TNFis) in the treatment of RA, but no randomized clinical trials have compared the three agents. Prior observational data are not consistent. We compared their effectiveness over 1 year in a prospective cohort. Analyses were performed on subjects' first episode of TNFi use in the Rheumatic Diseases Portuguese Register, Reuma.pt. The primary outcome was the proportion of patients with European League Against Rheumatism good response sustained at two consecutive observations separated by 3 months during the first year of TNFi use. Comparisons were performed using conventional adjusted logistic regression, as well as matching subjects across the three agents using a propensity score. In addition, baseline predictors of treatment response to TNFi were identified. The study cohort included 617 RA patients, 250 starting etanercept, 206 infliximab and 161 adalimumab. Good response was achieved by 59.6% for adalimumab, 59.2% for etanercept and 51.9% for infliximab (P = 0.21). The modelled probability of good response did not significantly differ across agents (etanercept vs adalimumab OR = 0.97, 95% CI 0.55, 1.71; etanercept vs infliximab OR = 1.25, 95% CI 0.74, 2.12; infliximab vs adalimumab OR = 0.80, 95% CI 0.47, 1.36). Matched propensity score analyses also showed no significant treatment response differences. Greater educational attainment was a predictor of better response, while smoking, presence of ACPA, glucocorticoid use and worse physician assessment of disease activity at baseline each predicted a reduced likelihood of treatment response. Over 1 year, we found no difference in effectiveness between adalimumab, etanercept and infliximab.


Problem-solving skills training for mothers of children with newly diagnosed cancer: a randomized trial.
Mothers of children with serious illnesses have lower levels of well-being than mothers in the general population. Problem-solving therapy (PST), a cognitive-behavioral intervention, has been shown to be effective in treating negative affectivity (depression, anxiety) and other manifestations of reduced well-being. This report describes a problem-solving skills training (PSST) intervention, based on problem-solving therapy, for mothers of newly diagnosed pediatric cancer patients. Ninety-two mothers were randomly assigned to receive PSST or to receive standard psychosocial care (Control Group). After the 8-week intervention, mothers in the PSST Group had significantly enhanced problem-solving skills and significantly decreased negative affectivity compared with controls. Analysis revealed that changes in self-reports of problem-solving behaviors accounted for 40% of the difference in mood scores between the two groups. Interestingly, PSST had the greatest impact on improving constructive problem solving, whereas improvement in mood was most influenced by decreases in dysfunctional problem solving. The implications of these findings for refinement of the PSST intervention and for extension to other groups of children with serious illnesses are discussed.


Pseudomonas otitidis sp. nov., isolated from patients with otic infections.
A novel Pseudomonas species, for which the name Pseudomonas otitidis sp. nov. is proposed, was identified from clinical specimens of infected human ears. Forty-one pseudomonads (34 from patients with acute otitis externa, six from patients with acute otitis media with otorrhoea and one from a patient with chronic suppurative otitis media) were recovered that did not match any known species. On the basis of genetic analyses and biochemical characterization, these isolates were shown to belong to the genus Pseudomonas. 16S rRNA gene sequence analysis and DNA-DNA hybridization studies indicated that this novel bacterium is closely related to, but different from, Pseudomonas aeruginosa. A description of this species is based on polyphasic studies of 11 clinical isolates. The type strain of Pseudomonas otitidis is MCC10330T (=ATCC BAA-1130T = DSM 17224T).


An optimized method for in situ hybridization with signal amplification that allows the detection of rare mRNAs.
In situ hybridization (ISH) using nonradioactive probes enables mRNAs to be detected with improved cell resolution but compromised sensitivity compared to ISH with radiolabeled probes. To detect rare mRNAs, we optimized several parameters for ISH using digoxygenin (DIG)-labeled probes, and adapted tyramide signal amplification (TSA) in combination with alkaline phosphatase (AP)-based visualization. This method, which we term TSA-AP, achieves the high sensitivity normally associated with radioactive probes but with the cell resolution of chromogenic ISH. Unlike published protocols, long RNA probes (up to 2.61 kb) readily permeated cryosections and yielded stronger hybridization signals than hydrolyzed probes of equivalent complexity. RNase digestion after hybridization was unnecessary and led to a substantial loss of signal intensity without significantly reducing nonspecific background. Probe concentration was also a key parameter for improving signal-to-noise ratio in ISH. Using these optimized methods on rat taste tissue, we detected mRNA for mGluR4, a receptor, and transducin, a G-protein, both of which are expressed at very low abundance and are believed to be involved in chemosensory transduction. Because the effect of the tested parameters was similar for ISH on sections of brain and tongue, we believe that these methodological improvements for detecting rare mRNAs may be broadly applicable to other tissues. (J Histochem Cytochem 47:431-445, 1999)


On the behaviour of biradicaloid [P(μ-NTer)]2 towards Lewis acids and bases.
The well-known diphosphadiazane-1,3-diyl [P(μ-NTer)]2 (Ter = 2,6-bis(2,4,6-trimethyl-phenyl)-phenyl) was treated with Lewis bases such as N-heterocyclic carbenes and Lewis acids e.g. gold(i) chloride complexes. In the reaction with the Lewis base, fragmentation of the P2N2 framework was observed, yielding a salt of the type [(NHC)2P](+)[(TerN)2P](-) in a clean reaction. The reaction of [P(μ-NTer)]2 with gold(i) chloride afforded 1 : 1 and 1 : 2 complexes. The dinuclear complex [(ClAu)2P(μ-NTer)2P] displays a bridging P atom between both gold centers, as has been observed for P based zwitterions.


Transcriptome Analysis Reveals Differential Expression of Genes Regulating Hepatic Triglyceride Metabolism in Pekin Ducks During Dietary Threonine Deficiency.
Dietary threonine (Thr) deficiency increases hepatic triglyceride accumulation in Pekin ducks, which results in fatty liver disease and impairs hepatic function. However, the underlying molecular mechanisms altered by dietary Thr deficiency are still unknown. To identify the underlying molecular changes, 180 one-day-old ducklings were divided into three groups, including Thr deficiency group (Thr-D), Thr sufficiency group (Thr-S), and pair-fed group (Pair-F) that was fed with a Thr-sufficient diet but with reduced daily feed intake. The results showed that feed intake was similar between Thr-D and Pair-F groups, but weight gain rate and final body weight in the Thr-D group were lower than those in the Pair-F group. Feed intake, weight gain, and body weight in Thr-D and Pair-F groups were lower than those in the Thr-S group. The Thr-D diet reduced abdominal fat percentage but increased hepatic triglyceride content when compared with that of the Thr-S and Pair-F groups. The Pair-F reduced hepatic levels of C15:0, C17:0, C18:0, C20:0, C20:4n6, and C22:0 and also reduced total fatty acid, saturated fatty acid, and unsaturated fatty acid content when compared with those of the Thr-D and Thr-S groups. The Thr-D diet increased hepatic content of C6:0, C17:1, C18:3n6, C20:0, C20:1n9, and C22:2, as well as reduced the content of C18:2n6t and C23:0 when compared with those of the Thr-S group. Transcriptome analysis in the liver indicated that the Thr-D diet upregulated genes related to fatty acid and triglyceride synthesis and downregulated genes related to fatty acid oxidation and triglyceride transport. Gene ontology analysis showed that more genes related to lipid metabolism processes and molecular function were differentially expressed in the Thr-D group relative to Thr-S and Pair-F groups than in the Pair-F group relative to the Thr-S group. KEGG pathway analysis showed that differentially expressed genes were enriched in signal transduction, immune, hormone, lipid, and amino acid metabolism pathways. Our findings indicated that the Thr-D diet increased hepatic triglyceride and fatty acid accumulation via increasing fatty acid and triglyceride synthesis and reducing fatty acid oxidation and triglyceride transport. These findings provide novel insights into our understanding of the molecular mechanisms underlying fat accumulation in the liver caused by dietary threonine deficiency.


Overabundance of rare-cutting restriction endonuclease sites in the human genome.
A human chromosome 3-specific cosmid library was constructed from a somatic cell hybrid containing human chromosome 3 as its only human component. This library was screened to identify 230 human recombinants which contained an average insert size of 37 kilobases. DNA prepared from 54 of these cosmids, representing 2000 kilobases of human DNA, was then tested for restriction endonuclease sites for EcoRI, HindIII, KpnI, XhoI, and DraI, as well as those of the rare-cutting restriction endonucleases NotI, SfiI, NruI, MluI, SacII, and BssHII. Sites for the latter enzymes were much more abundant than would be expected from theoretical calculations, reflecting non-random clustering of these sites. This has important implications for the use of these enzymes in the construction of physical maps of chromosomes. Some individual cosmids contained large numbers of rare sites, offering an alternative means of physically mapping chromosomes based upon identifying clusters of rare restriction sites. These clusters appear to be spaced an average of 1000 kb apart.


Translational regulation of mRNAs for ribosomal proteins during early Drosophila development.
In Drosophila, the vast majority of mRNAs that are polysome associated during oogenesis are also polysome associated during early embryogenesis. We have previously identified an exceptional mRNA that appears to be depleted from early-embryo polysomes [Fruscoloni, P., Al-Atia, G. R., & Jacobs-Lorena, M. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 3359-3363]. This mRNA has been subsequently identified as coding for a ribosomal protein (r-protein) [Kay, M., & Jacobs-Lorena, M. (1985) Mol. Cell. Biol. (in press)]. Changes in association with polysomes of two r-protein mRNAs during early Drosophila development were investigated for this report. Hybridization of cloned DNA probes to blots of RNA obtained from sucrose gradient fractions reveals that r-protein mRNAs are substantially associated with polysomes during oogenesis, depleted from polysomes during early embryogenesis, and again polysome associated during late embryogenesis. Thus, translation of r-protein mRNAs parallels transcription of ribosomal RNA (rRNA) during this time of development. By contrast, no such differences were observed when actin and histone probes were used as controls and hybridized to the same blots. The abundance of mRNAs for r-proteins as a function of development was also measured. Abundance was relatively high and constant during oogenesis and embryogenesis (when translational regulation is apparent), somewhat decreased in larval and pupal stages, and low in adult nonovarian tissues. Coordination between r-protein and rRNA synthesis appears to be achieved by regulating translation of r-protein mRNAs in early embryos and by decreasing their abundance in adult tissues.


Taximin, a conserved plant-specific peptide is involved in the modulation of plant-specialized metabolism.
Small peptides play important roles in the signalling cascades that steer plant growth, development and defence, and often crosstalk with hormonal signalling. Thereby, they also modulate metabolism, including the production of bioactive molecules that are of high interest for human applications. Yew species (Taxus spp.) produce diterpenes such as the powerful anticancer agent paclitaxel, the biosynthesis of which can be stimulated by the hormone jasmonate, both in whole plants and cell suspension cultures. Here, we identified Taximin, as a gene encoding a hitherto unreported, plant-specific, small, cysteine-rich signalling peptide, through a transcriptome survey of jasmonate-elicited T. baccata suspension cells grown in two-media cultures. Taximin expression increased in a coordinated manner with that of paclitaxel biosynthesis genes. Tagged Taximin peptides were shown to enter the secretory system and localize to the plasma membrane. In agreement with this, the exogenous application of synthetic Taximin peptide variants could transiently modulate the biosynthesis of taxanes in T. baccata cell suspension cultures. Importantly, the Taximin peptide is widely conserved in the higher plant kingdom with a high degree of sequence conservation. Accordingly, Taximin overexpression could stimulate the production of nicotinic alkaloids in Nicotiana tabacum hairy root cultures in a synergistic manner with jasmonates. In contrast, no pronounced effects of Taximin overexpression on the specialized metabolism in Medicago truncatula roots were observed. This study increases our understanding of the regulation of Taxus diterpene biosynthesis in particular and plant metabolism in general. Ultimately, Taximin might increase the practical potential of metabolic engineering of medicinal plants.


Embryonic response to long-term exposure of the marine crustacean Nephrops norvegicus to ocean acidification and elevated temperature.
Due to anthropogenic CO2 emissions, our oceans have gradually become warmer and more acidic. To better understand the consequences of this, there is a need for long-term (months) and multistressor experiments. Earlier research demonstrates that the effects of global climate change are specific to species and life stages. We exposed berried Norway lobsters (Nephrops norvegicus), during 4 months to the combination of six ecologically relevant temperatures (5-18°C) and reduced pH (by 0.4 units). Embryonic responses were investigated by quantifying proxies for development rate and fitness including: % yolk consumption, mean heart rate, rate of oxygen consumption, and oxidative stress. We found no interactions between temperature and pH, and reduced pH only affected the level of oxidative stress significantly, with a higher level of oxidative stress in the controls. Increased temperature and % yolk consumed had positive effects on all parameters except on oxidative stress, which did not change in response to temperature. There was a difference in development rate between the ranges of 5-10°C (Q 10: 5.4) and 10-18°C (Q 10: 2.9), implicating a thermal break point at 10°C or below. No thermal limit to a further increased development rate was found. The insensitivity of N. norvegicus embryos to low pH might be explained by adaptation to a pH-reduced external habitat and/or internal hypercapnia during incubation. Our results thus indicate that this species would benefit from global warming and be able to withstand the predicted decrease in ocean pH in the next century during their earliest life stages. However, future studies need to combine low pH and elevated temperature treatments with hypoxia as hypoxic events are frequently and increasingly occurring in the habitat of benthic species.


Steatosis and NASH in type 2 diabetes.
Non Alcoholic Fatty Liver Disease (NAFLD) is currently the most common chronic liver disease in the world, encompassing various conditions ranging from simple steatosis, steatohepatitis, to fibrosis and cirrhosis. The association between NAFLD and Type 2 Diabetes (T2D) is strong and complex, given that the prevalence of NAFLD is particularly high in individuals with Type 2 Diabetes. In fact, insulin resistance occurring in this metabolic disease can promote NAFLD development, and vice versa, NAFLD can enhance insulin resistance. In this review, we focus on the mechanisms linking NAFLD and T2D, including fatty acid accumulation, inflammation, oxidative stress etc. We also discuss about situations showing a dissociation between steatosis and insulin resistance, in order to provide new insights for NAFLD therapeutic targets.


[Applicability of the Torrington and Henderson scale].
To validate the Torrington & Henderson preoperative risk assessment program in the population of surgical patients in an university hospital (Hospital São Paulo, UNIFESP, São Paulo, Brasil). We evaluated 1162 patients who underwent to major thoracic, upper and under abdominal surgery using the Torrington & Henderson program. The patients were classified in low (785), moderate (317) and high risk (60). All patients were accompanied daily during the postoperative period by the same medical team which assessed the preoperative period, until discharged or died. The postoperative pulmonary complications were present in 6.1%, 23.3% and 35.0% in the low, moderate and high risk respectively. The relative risk to develop postoperative pulmonary complications was 3.8 higher in the patients with moderate risk and 5.7 higher in the patients with high risk. The mortality rate due to pulmonary complications was 1.7%, 6.3% and 11.7% respectively in the patients with low, moderate and high risk. The relative risk to the death was 3.7 higher in the patients with moderate risk and 6.9 in the high risk. We concluded that the Torrington & Henderson preoperative risk assessment program can identify in our population patients who will develop postoperative pulmonary complications.


Doxycycline versus azithromycin for treatment of leptospirosis and scrub typhus.
Leptospirosis and scrub typhus are important causes of acute fever in Southeast Asia. Options for empirical therapy include doxycycline and azithromycin, but it is unclear whether their efficacies are equivalent. We conducted a multicenter, open, randomized controlled trial with adult patients presenting with acute fever (<15 days), without an obvious focus of infection, at four hospitals in Thailand between July 2003 and January 2005. Patients were randomly allocated to receive either a 7-day course of doxycycline or a 3-day course of azithromycin. The cure rate, fever clearance time, and adverse drug events were compared between the two study groups. A total of 296 patients were enrolled in the study. The cause of acute fever was determined for 151 patients (51%): 69 patients (23.3%) had leptospirosis; 57 patients (19.3%) had scrub typhus; 14 patients (4.7%) had murine typhus; and 11 patients (3.7%) had evidence of both leptospirosis and a rickettsial infection. The efficacy of azithromycin was not inferior to that of doxycycline for the treatment of both leptospirosis and scrub typhus, with comparable fever clearance times in the two treatment arms. Adverse events occurred more frequently in the doxycycline group than in the azithromycin group (27.6% and 10.6%, respectively; P = 0.02). In conclusion, doxycycline is an affordable and effective choice for the treatment of both leptospirosis and scrub typhus. Azithromycin was better tolerated than doxycycline but is more expensive and less readily available.


Neurologic manifestations in hospitalized patients with COVID-19: The ALBACOVID registry.
The coronavirus disease 2019 (COVID-19) has spread worldwide since December 2019. Neurologic symptoms have been reported as part of the clinical spectrum of the disease. We aimed to determine whether neurologic manifestations are common in hospitalized patients with COVID-19 and to describe their main characteristics. We systematically reviewed all patients diagnosed with COVID-19 admitted to the hospital in a Spanish population during March 2020. Demographic characteristics, systemic and neurologic clinical manifestations, and complementary tests were analyzed. Of 841 patients hospitalized with COVID-19 (mean age 66.4 years, 56.2% men), 57.4% developed some form of neurologic symptom. Nonspecific symptoms such as myalgias (17.2%), headache (14.1%), and dizziness (6.1%) were present mostly in the early stages of infection. Anosmia (4.9%) and dysgeusia (6.2%) tended to occur early (60% as the first clinical manifestation) and were more frequent in less severe cases. Disorders of consciousness occurred commonly (19.6%), mostly in older patients and in severe and advanced COVID-19 stages. Myopathy (3.1%), dysautonomia (2.5%), cerebrovascular diseases (1.7%), seizures (0.7%), movement disorders (0.7%), encephalitis (n = 1), Guillain-Barré syndrome (n = 1), and optic neuritis (n = 1) were also reported, but less frequent. Neurologic complications were the main cause of death in 4.1% of all deceased study participants. Neurologic manifestations are common in hospitalized patients with COVID-19. In our series, more than half of patients presented some form of neurologic symptom. Clinicians need to maintain close neurologic surveillance for prompt recognition of these complications. The mechanisms and consequences of severe acute respiratory syndrome coronavirus type 2 neurologic involvement require further studies.


[Phosphorylase and gluco-6-phosphatase activity and glycogen level in tissues of vertebrates].
The activity of phosphorylase (EC 22.214.171.124), glucose-6-phosphatase (EC 22.214.171.124) and the content of glycogen have been determined in tissues of fish, amphibians, reptiles, mammalians. No differences in the activity of phosphorylase and glucose-6-phosphatase in the liver, myocardium, and brain of animals of the phylogenetic groups under study are found. The activity of glucose-6-phosphatase in the anaerobic muscles of poikilothermal animals is found to be rather high. The share of phosphorylase a in the skeletal muscles and brain as well as the glycogen content in the brain of these animals is essentially higher than that of adult mammalians.


Prepubertal OVX increases IGF-I expression and bone accretion in C57BL/6J mice.
It is generally well accepted that the pubertal surge in estrogen is responsible for the rapid bone accretion that occurs during puberty and that this effect is mediated by an estrogen-induced increase in growth hormone (GH)/insulin-like growth factor (IGF) action. To test the cause and effect relationship between estrogen and GH/IGF, we evaluated the consequence of ovariectomy (OVX) in prepubertal mice (C57BL/6J mice at 3 wk of age) on skeletal changes and the GH/IGF axis during puberty. Contrary to our expectations, OVX increased body weight (12-18%), bone mineral content (11%), bone length (4%), bone size (3%), and serum, liver, and bone IGF-I (30-50%) and decreased total body fat (18%) at 3 wk postsurgery. To determine whether estrogen is the key ovarian factor responsible for these changes, we performed a second experiment in which OVX mice were treated with placebo or estrogen implants. In addition to observing similar results compared with our first experiment, estrogen treatment partially rescued the increased body weight and bone size and completely rescued body fat and IGF-I levels. The increased bone accretion in OVX mice was due to increased bone formation rate (as determined by bone histomorphometry) and increased serum procollagen peptide. In conclusion, contrary to the known estrogen effect as an initiator of GH/IGF surge and thereby pubertal growth spurt, our findings demonstrate that loss of estrogen and/or other hormones during the prepubertal growth period effect leads to an increase in IGF-I production and bone accretion in mice.


Mathematical algorithm for the automatic recognition of intestinal parasites.
Parasitic infections are generally diagnosed by professionals trained to recognize the morphological characteristics of the eggs in microscopic images of fecal smears. However, this laboratory diagnosis requires medical specialists which are lacking in many of the areas where these infections are most prevalent. In response to this public health issue, we developed a software based on pattern recognition analysis from microscopi digital images of fecal smears, capable of automatically recognizing and diagnosing common human intestinal parasites. To this end, we selected 229, 124, 217, and 229 objects from microscopic images of fecal smears positive for Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica, respectively. Representative photographs were selected by a parasitologist. We then implemented our algorithm in the open source program SCILAB. The algorithm processes the image by first converting to gray-scale, then applies a fourteen step filtering process, and produces a skeletonized and tri-colored image. The features extracted fall into two general categories: geometric characteristics and brightness descriptions. Individual characteristics were quantified and evaluated with a logistic regression to model their ability to correctly identify each parasite separately. Subsequently, all algorithms were evaluated for false positive cross reactivity with the other parasites studied, excepting Taenia sp. which shares very few morphological characteristics with the others. The principal result showed that our algorithm reached sensitivities between 99.10%-100% and specificities between 98.13%- 98.38% to detect each parasite separately. We did not find any cross-positivity in the algorithms for the three parasites evaluated. In conclusion, the results demonstrated the capacity of our computer algorithm to automatically recognize and diagnose Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica with a high sensitivity and specificity.


The peculiarity of the metal-ceramic interface.
Important properties of materials are strongly influenced or controlled by the presence of solid interfaces, i.e. from the atomic arrangement in a region which is a few atomic spacing wide. Using the quantitative analysis of atom column positions enabled by CS-corrected transmission electron microscopy and theoretical calculations, atom behaviors at and adjacent to the interface was carefully explored. A regular variation of Cu interplanar spacing at a representative metal-ceramic interface was experimentally revealed, i.e. Cu-MgO (001). We also found the periodic fluctuations of the Cu and Mg atomic positions triggered by the interfacial geometrical misfit dislocations, which are partially verified by theoretical calculations using empirical potential approach. Direct measurements of the bond length of Cu-O at the coherent regions of the interface showed close correspondence with theoretical results. By successively imaging of geometrical misfit dislocations at different crystallographic directions, the strain fields around the interfacial geometrical misfit dislocation are quantitatively demonstrated at a nearly three-dimensional view. A quantitative evaluation between the measured and calculated strain fields using simplified model around the geometrical misfit dislocation is shown.


Sandwich enzyme-linked immunosorbent assay (ELISA) for detection of mustard in foods.
Undeclared mustard residues in food products could trigger allergic reactions in mustard-allergic consumers. Our objective was to develop and validate a sandwich-type ELISA for the detection of mustard residues in foods. A mixture of yellow, brown, and oriental mustard seeds was used to immunize 3 rabbits and 1 sheep. Two mustard ELISAs were developed by utilizing the reciprocal combination of rabbit and sheep polyclonal antimustard sera as the capture and detector reagents. Binding was visualized by addition of rabbit antisheep or goat antirabbit IgG antibody labeled with alkaline phosphatase and subsequent addition of substrate. The optimized ELISAs have limits of quantification (LOQ) of 1 and 3 ppm (mug of ground, whole mustard seeds/mL) for the sheep capture and rabbit capture formats, respectively. Only rapeseed cross-reacted in the rabbit and sheep capture mustard ELISAs at a level equivalent to 12300 and 16900 ppm of mustard. The mean percent recovery for cooked frankfurters spiked with 0 to 1000 ppm mustard flour was 95.3%+/- 10.7%. A limited retail survey of 29 foods revealed that, of 15 samples having mustard declared on the ingredient list, 2 baked bean products contained no detectable mustard, possibly owing to a decrease in extractability and detectability of mustard proteins after subjecting to thermal processing. For the remaining 14 samples without mustard declared on the label, 3 samples contained detectable mustard, presumably due to the labeling of mustard as "spice" or inadvertent cross-contamination. This sandwich-type ELISA can serve as a powerful tool for food manufacturers and regulatory agencies to detect and quantify mustard residues in processed foods.


[Quality of life of pregnant women in homeopathic or mainstream medical type of care and the course of the pregnancy].
This study investigated 120 pregnant women cared for and treated by physicians specialized in homeopathy and 85 pregnant women cared for and treated by mainstream gynecologists. Quality of life was assessed twice during pregnancy and once shortly after delivery by the instrument SEIQoL (Schedule for the Evaluation of Individualized Quality of Life). In addition, the study registered individual personality characteristics, the psychological situation, the attitude towards the forthcoming delivery, and the functional state before and after delivery. It turned out that women who preferred homeopathic care judged their quality of life lower than women from the other group. However, their criteria for medical decisions and for the assessment of their own personal situation differed as well. There was some correlation between quality of life and age, number of deliveries, partnership and certain personality characteristics. The study showed the importance of paying more attention to the subjective world of the pregnant women. This applies especially in an analysis of two dissimilar therapeutic systems. It could also be concluded that the SEIQoL was a useful instrument for such a study. Finally it turned out that the two populations investigated were different.


Three month inhalation exposure to low-level PM2.5 induced brain toxicity in an Alzheimer's disease mouse model.
Although numerous epidemiological studies revealed an association between ambient fine particulate matter (PM2.5) exposure and Alzheimer's disease (AD), the PM2.5-induced neuron toxicity and associated mechanisms were not fully elucidated. The present study assessed brain toxicity in 6-month-old female triple-transgenic AD (3xTg-AD) mice following subchronic exposure to PM2.5 via an inhalation system. The treated mice were whole-bodily and continuously exposed to real-world PM2.5 for 3 months, while the control mice inhaled filtered air. Changes in cognitive and motor functions were evaluated using the Morris Water Maze and rotarod tests. Magnetic resonance imaging analysis was used to record gross brain volume alterations, and tissue staining with hematoxylin and eosin, Nissl, and immunohistochemistry methods were used to monitor pathological changes in microstructures after PM2.5 exposure. The levels of AD-related hallmarks and the oxidative stress biomarker malondialdehyde (MDA) were assessed using Western blot analysis and liquid chromatography-mass spectrometry, respectively. Our results showed that subchronic exposure to environmental levels of PM2.5 induced obvious neuronal loss in the cortex of exposed mice, but without significant impairment of cognitive and motor function. Increased levels of phosphorylated-tau and MDA were also observed in olfactory bulb or hippocampus after PM2.5 exposure, but no amyloid pathology was detected, as reported in previous studies. These results revealed that a relatively lower level of PM2.5 subchronic exposure from the environmental atmosphere still induced certain neurodegenerative changes in the brains of AD mice, especially in the olfactory bulb, entorhinal cortex and hippocampus, which is consistent with the nasal entry and spreading route for PM exposure. Systemic factors may also contribute to the neuronal toxicity. The effects of PM2.5 after a more prolonged exposure period are needed to establish a more comprehensive picture of the PM2.5-mediated development of AD.


3D Facial morphology in children affected by spinal muscular atrophy type 2 (SMAII).
The main objective of this study was to assess the three-dimensional facial characteristics of children affected by spinal muscular atrophy (SMA), a severe muscular disorder characterized by hypotonia, areflexia, weakness, and respiratory impairment. Stereophotogrammetric facial scans from 22 SMA type II patients aged 2-7 years were obtained. Data were analysed using both inter-landmark distances and principal component analysis and compared with data collected from matched control subjects. Patients had wider transverse facial diameters, but smaller biocular width. Middle and lower anterior face heights were increased, whereas the mandibular ramus was shorter, with a reduced posterior-to-anterior face height ratio. Facial width-to-length ratio was reduced. In the sagittal plane, mandibular body length, and facial divergence were increased, whereas the gonial angles were decreased. In the horizontal plane, lower facial convexity was greater in patients, whereas mandibular convexity was smaller. Patients had smaller and down-slanted eye fissures, with a larger and more vertically developed nose. This study assessed a relatively small number of patients, due to the rare frequency of SMA type II. SMA type II children possess peculiar facial alterations that may be due to the altered muscular activity. As feeding problems may derive also by malocclusion and masticatory muscular alterations, a detailed assessment of the craniofacial individual alterations should be considered in the standards of care of these patients.


[Family experience on the care needs relative to insulin therapy and diabetic foot prevention].
This study aimed at describing routine insulin therapy and lower-limb lesion prevention practices used by relatives of diabetes mellitus patients. Semi-structured interviews were carried out at home, and 38 families of the municipal district of Bandeirantes, Paraná, Brazil, were included. Results revealed that relatives had good knowledge on glycemic control and diabetic foot prevention, particularly of non-medication care, with emphasis on food. Insulin therapy care is very demanding for relatives, both in emotional and technical terms. It was concluded that the quality of the relationship among family members, encouraging the patient to comply with insulin therapy and special diet, and concern with food items available at home are essential for the patient with diabetes mellitus.


aly/aly mice: a unique model of biliary disease.
An autosomal recessive murine mutation, coined "aly/aly" or "alymphoplasia," was recently reported. Homozygotes for aly are defective in both humoral and cell-mediated immune function and have diffuse lymphoid cell infiltration of various tissues, particularly around the conduit ducts of the pancreas and salivary glands. In pilot studies in our laboratories, aly/aly mice were found to have peculiar biliary tract lesions, which were analyzed histologically and immunohistochemically in the present study. The livers of aly/aly mice older than 8 weeks consistently showed a variable lymphoid cell infiltration with lymph follicle formation in portal tracts; intrahepatic biliary epithelial cells showed various types of damage including pseudopyloric gland metaplasia and proliferative changes. In addition, the extrahepatic bile duct and intrahepatic large bile duct were found to contain an acidophilic substance in their epithelial cytoplasm. In the lumen and occasionally in the cytoplasm of these bile ducts, acidophilic crystals were also seen. Ultrastructurally, the intracytoplasmic acidophilic substances consisted of membrane-bound intracytoplasmic inclusions with homogeneous electron density, likely derived from rough endoplasmic reticulum (ER). Immunohistochemically, the cytoplasmic acidophilic substances were simultaneously positive for cystatin C, gastrin, serotonin, and somatostatin. In contrast, the acidophilic crystals did not react with any of these antibodies. These findings suggest that the intracytoplasmic acidophilic substances may contain a precursor of the peptide hormones, possibly because of defective secretion or intracellular transport. We believe that the aly/aly mouse is a useful model for the analysis of biliary metabolic events, and for studies of the interaction of the immune system and biliary destruction.


Controlled experiments measuring personal exposure to PM2.5 in close proximity to cigarette smoking.
Few measurements of exposure to secondhand smoke (SHS) in close proximity to a smoker are available. Recent health studies have demonstrated an association between acute (<2 h) exposures to high concentrations of SHS and increased risk of cardiovascular and respiratory disease. We performed 15 experiments inside naturally ventilated homes and 16 in outdoor locations, each with 2-4 non-smokers sitting near a cigarette smoker. The smoker's and non-smokers' real-time exposures to PM2.5 from SHS were measured by using TSI SidePak monitors to sample their breathing zones. In 87% of the residential indoor experiments, the smoker received the highest average exposure to SHS, with PM2.5 concentrations ranging from 50-630 μg/m(3) . During the active smoking period, individual non-smokers sitting within approximately 1 m of a smoker had average SHS exposures ranging from negligible up to >160 μg/m(3) of PM2.5 . The average incremental exposure of the non-smokers was higher indoors (42 μg/m(3) , n = 35) than outdoors (29 μg/m(3) , n = 47), but the overall indoor and outdoor frequency distributions were similar. The 10-s PM2.5 averages during the smoking periods showed great variability, with multiple high concentrations of short duration (microplumes) both indoors and outdoors.


[Age and gender features of tobacco smoking].
On the basis of the conducted research, prevalence of tobacco smoking and its intensity are described as well as dependence on gender and age in an urban population of the middle Volga region (on the example of the city of Samara). It is proved, that in a general selection among men and women smoke: 49,37 and 14,17%; smoked before: 22,80 and 8,86%; never smoked: 27,83 and 76,97% of inspected men and women, accordingly. The contribution of the education level and marriage status are set in epidemiology of tobacco smoking dependence. Authors consider the development of public and medical measures in a fight against tobacco smoking should be realized, taking into account the gender, age, social and mental features of population, which is a difficult, but necessary process.


The cost of smoking in Canada, 1991.
In 1991, smoking-attributable health care costs in Canada were $2.5 billion (CAN). Additional smoking-attributable costs included $1.5 billion for residential care, $2 billion due to workers' absenteeism, $80 million due to fires and $10.5 billion due to lost future income caused by premature death. Adjustments for future costs if smoking had not occurred and smokers had not died were estimated to be $1.5 billion. According to this analysis, smokers cost society about $15 billion while contributing roughly $7.8 billion in taxes. The results indicate that smoking-attributable costs in Canada have increased steadily since 1966 to the 1991 value of $15 billion. Nevertheless, while the determination of smoking-attributable costs is important, the issue continues to be public health. In addition, for the first time in Canada, the smoking-attributable cost for residential care has been estimated.


Generalized Maxwell projections for multi-mode network Photonics.
The design of optical resonant systems for controlling light at the nanoscale is an exciting field of research in nanophotonics. While describing the dynamics of few resonances is a relatively well understood problem, controlling the behavior of systems with many overlapping states is considerably more difficult. In this work, we use the theory of generalized operators to formulate an exact form of spatio-temporal coupled mode theory, which retains the simplicity of traditional coupled mode theory developed for optical waveguides. We developed a fast computational method that extracts all the characteristics of optical resonators, including the full density of states, the modes quality factors, and the mode resonances and linewidths, by employing a single first principle simulation. This approach can facilitate the analytical and numerical study of complex dynamics arising from the interactions of many overlapping resonances, defined in ensembles of resonators of any geometrical shape and in materials with arbitrary responses.


Endoscopic sleeve gastroplasty: results from a single surgical bariatric centre.
The aim of this study was to evaluate the safety and efficacy of the endoscopic sleeve gastroplasty (ESG) procedure. Patients ineligible for bariatric surgery due to comorbidities or low Body Mass Index (BMI) were offered ESG. Gastric tubularization was carried out via multiple multi-bite sutures across the greater curvature of the stomach. The patients underwent a water-soluble swallow test on post-operative day 1 (POD-1) to assess gastric emptying and were placed on a soft diet if upper GI tract function was confirmed. From January 2019 to March 2022, 27 patients underwent ESG: 14 for severe obesity with comorbidities, including liver transplant, end-stage kidney disease, severe cardiovascular and respiratory diseases. The mean BMI before treatment was 36 ± 9 kg/m2. Two patients (7%) who developed gastric bleeding were successfully treated with packed red blood cells (PRBC) transfusions. After a mean follow-up of 18 months, the percentage of total body weight loss (%TBWL) and the percentage of excess weight (%EWL) were 11 ± 7 and 39 ± 27, respectively. The latter was significantly higher in the patients with an initial BMI < 40 kg/m2 (50 vs 22, p < 0.05). The patients whose gastric sleeve extended for more than a third of the length of the stomach (p < 0.05) had better results. ESG was found to be effective and safe in high-risk surgical patients whose initial BMI was (< 40). Studies characterized by larger number of patients and longer follow-up periods will be able to confirm these results.


National patterns of care for pancreatic cancer. Results of a survey by the Commission on Cancer.
The Commission on Cancer of the American College of Surgeons conducted a large, national survey to assess methods of diagnosis, American Joint Commission on Cancer staging, treatment, and outcome of patients with adenocarcinoma of the pancreas. The survey questionnaire contained 160 questions and covered two study periods, 1983 to 1985 and 1990, for time-trend analysis. Nine hundred seventy-eight institutions throughout the United States voluntarily participated, contributing 8917 case reports for 1983 to 1985 and 8025 reports for 1990, resulting in a total of 16,942 patient reports. Most, but not all, of the participating hospitals maintain approval status with the Commission on Cancer of the American College of Surgeons. The ratio of male-to-female cases was 1:1. Patient characteristics including age, ethnicity, neighborhood income, type of insurance coverage, and hospital characteristics--including annual caseload and type of facility (e.g., teaching, community)--appeared to influence surgical multimodality treatment patterns. The most common presenting symptom was abdominal pain. The reported history of smoking for these patients with pancreatic cancer was higher than U.S. population averages. The frequency of using abdominal computed tomography scans, endoscopic retrograde cholangiopancreatography, carcinoembryonic antigen, and CA 19-9 during patient evaluation all increased. Time trends toward lower operative mortality and more extirpative surgery were reported, as was a slightly higher survival for those patients who were resected surgically. Pancreatic cancer continues to be a disease of older patients. There were slight improvements in operative mortality. For a highly selective category of patients, cancer-directed surgery offers a chance for cure with excellent operative mortality and acceptable complication rates, especially when performed in institutions that have a 20 or greater case per year experience.


Identification of Attenuators of Transcriptional Termination: Implications for RNA Regulation in Escherichia coli.
The regulatory function of many bacterial small RNAs (sRNAs) requires the binding of the RNA chaperone Hfq to the 3' portion of the sRNA intrinsic terminator, and therefore sRNA signaling might be regulated by modulating its terminator. Here, using a multicopy screen developed with the terminator of sRNA SgrS, we identified an sRNA gene (cyaR) and three protein-coding genes (cspD, ygjH, and rof) that attenuate SgrS termination in Escherichia coli. Analyses of CyaR and YgjH, a putative tRNA binding protein, suggested that the CyaR activity was indirect and the effect of YgjH was moderate. Overproduction of the protein attenuators CspD and Rof resulted in more frequent readthrough at terminators of SgrS and two other sRNAs, and regulation by SgrS of target mRNAs was reduced. The effect of Rof, a known inhibitor of Rho, was mimicked by bicyclomycin or by a rho mutant, suggesting an unexpected role for Rho in sRNA termination. CspD, a member of the cold shock protein family, bound both terminated and readthrough transcripts, stabilizing them and attenuating termination. By RNA sequencing analysis of the CspD overexpression strain, we found global effects of CspD on gene expression across some termination sites. We further demonstrated effects of endogenous CspD under slow growth conditions where cspD is highly expressed. These findings provided evidence of changes in the efficiency of intrinsic termination, confirming this as an additional layer of the regulation of sRNA signaling. IMPORTANCE Growing evidence suggests that the modulation of intrinsic termination and readthrough of transcription is more widespread than previously appreciated. For small RNAs, proper termination plays a critical role in their regulatory function. Here, we present a multicopy screen approach to identify factors that attenuate small RNA termination and therefore abrogate signaling dependent on the small RNA. This study highlights a new aspect of regulation of small RNA signaling as well as the modulation of intrinsic termination.


Crossed corticospinal facilitation between arm and trunk muscles in humans.
A voluntary contraction of muscles with one arm increases the excitability of corticospinal projections to the contralateral resting arm, a phenomenon known as crossed facilitation. Although many motor tasks engage simultaneous activation of the arm and trunk, interactions between corticospinal projections targeting these segments remain largely unknown. Using transcranial magnetic stimulation over the trunk representation of the primary motor cortex, we examined motor-evoked potentials (MEPs) in the resting erector spinae (ES) muscle when the contralateral arm remained at rest or performed 20% of isometric maximal voluntary contraction (MVC) into index finger abduction, thumb abduction, elbow flexion, and elbow extension. We found that MEP size in the ES increased during all voluntary contractions, with greater facilitation occurring during elbow flexion and index finger abduction. To further examine the origin of changes in MEP size, we measured short-interval intracortical inhibition (SICI) and cervicomedullary MEPs (CMEPs) in the ES muscle during elbow flexion and index finger abduction and when the arm remained at rest. Notably, SICI decreased and CMEPs remained unchanged in the ES during both voluntary contractions compared with rest, suggesting a cortical origin for the effects. Our findings reveal crossed facilitatory interactions between trunk extensor and proximal and distal arm muscles, particularly for elbow flexor and index finger muscles, likely involving cortical mechanisms. These interactions might reflect the different role of these muscles during functionally relevant arm and trunk movements. NEW & NOTEWORTHY Many of the tasks of daily life involve simultaneous activation of the arm and trunk. We found that responses in the erector spinae muscles evoked by motor cortical stimulation increased in size during elbow flexion and extension and during index finger abduction and thumb abduction. Crossed facilitation with the trunk was more pronounced during elbow flexion and index finger abduction. These results might reflect the different role of these muscles during arm and trunk movements.


Evolutionary conservation and chromosomal localization of flvi-1.
A locus in feline DNA, termed flvi-1, has been identified as harboring retroviral integrations commonly found in natural feline lymphomas induced by infection with feline leukemia virus (FeLV). Southern blot analysis of human and murine DNA using restriction fragments representing flvi-1 demonstrates its phylogenetic conservation among mammals, flvi-1 is localized to murine chromosome 2, proximal portion of band E, by in situ hybridization to metaphase chromosomes. This position is adjacent to that of another putative proto-oncogene, sfpi-1, although probes representing flvi-1 and sfpi-1 do not cross-hybridize. The repeated implication of flvi-1 in natural feline leukemogenesis, its evolutionary conservation and its chromosomal position support the hypothesis that flvi-1 may represent a previously unidentified protooncogene.


Effect of some stressors on pathogenicity of Eimeria tenella in broiler chicken.
The present investigation was carried out to study the effect of stress on severity of Eimeria tenella infection in chickens. Stressors applied were vaccination and crowdness. One hundred and twenty five one-day old chicks were divided into five groups. The groups were as follow: group (1) control negative, group (2) control positive, group (3) crowded non-vaccinated, group (4) vaccinated, and group (5) vaccinated crowded. The chicks of 2-5 groups were inoculated with 10 x 10(3) sporulated oocysts of E. tenella per chick at 3rd day old, then challenged with 15 x 10(3) sporulated oocysts of E. tenella on day 28. The results revealed that, the highest mortality percent, oocysts production and caecal scores were recorded in the crowded vaccinated group. This group also exhibited the lowest weight of body, spleen, bursa and thymus. At the last four weeks, it was found that overstocking of chickens lead to elevation of corticosterone levels. Total proteins, glucose, calcium and phosphorus levels decreased in crowded vaccinated group.


Capillary pressures in rat intestinal muscle and mucosal villi during venous pressure elevation.
Whole-organ experiments designed to estimate the capillary filtration coefficient require information about the numerical relationship between capillary pressure and venous pressure. Indirect estimates using isogravimetric and isovolumetric methods indicate that 62-85% of a step change in venous pressure reaches the intestinal capillaries, taken as a whole. We have made direct measurements of capillary pressure with a servo-null micropressure system in the microcirculation of both the intestinal muscle and the mucosal villi of rats during local elevation of venous pressure. Consistent regional differences in the relationship between capillary pressure and venous pressure were observed. During increased venous pressure, submucosal arterioles constricted, while muscularis arterioles dilated. The diameter changes of the small arterioles were consistent with blood flow redistribution from mucosa to muscle during venous pressure elevation, but inconsistent with a pure myogenic response. These data raise questions about the exact role for the expression of the myogenic response during venous pressure elevation in the intestine and about previous interpretations of whole-organ experiments concerned with intestinal blood flow and fluid exchange.


Slovenian midwives' and nurses' views on post-natal depression: an exploratory study.
Post-natal depression affects approximately 10-15% of women. The literature suggests that midwives and nurses are key professionals in dealing with post-natal mood disorders. However, this would be a new role in Slovenia for which it is not clear whether midwives and nurses are prepared. This study explored Slovenian midwives' and nurses' knowledge of, and attitudes towards, post-natal mood disorders. Two focus groups were conducted, each with five participants, working in the maternity hospital and in the community centre in Postojna. Data were grouped into three main themes--knowledge of post-natal mental health disorders, role in the management of these issues, and perceived problems and possible solutions in the health care of post-natally depressed women. Participants lacked knowledge on post-natal mental health and did not consider its management to be their role. They saw the main obstacle to caring for these women as being a lack of continuity of carer. Slovenian midwives and nurses should be appropriately educated regarding this subject in order to become more capable of managing post-natal mental health. Continuous relationships with women should be encouraged. Further investigation is required and a questionnaire survey involving a larger sample of midwives and community nurses is planned on the basis of the findings of this study.