Developing tools for the safety specification in risk management plans: lessons learned from a pilot project.
Following the adoption of the ICH E2E guideline, risk management plans (RMP) defining the cumulative safety experience and identifying limitations in safety information are now required for marketing authorisation applications (MAA). A collaborative research project was conducted to gain experience with tools for presenting and evaluating data in the safety specification. This paper presents those tools found to be useful and the lessons learned from their use. Archive data from a successful MAA were utilised. Methods were assessed for demonstrating the extent of clinical safety experience, evaluating the sensitivity of the clinical trial data to detect treatment differences and identifying safety signals from adverse event and laboratory data to define the extent of safety knowledge with the drug. The extent of clinical safety experience was demonstrated by plots of patient exposure over time. Adverse event data were presented using dot plots, which display the percentages of patients with the events of interest, the odds ratio, and 95% confidence interval. Power and confidence interval plots were utilised for evaluating the sensitivity of the clinical database to detect treatment differences. Box and whisker plots were used to display laboratory data. This project enabled us to identify new evidence-based methods for presenting and evaluating clinical safety data. These methods represent an advance in the way safety data from clinical trials can be analysed and presented. This project emphasises the importance of early and comprehensive planning of the safety package, including evaluation of the use of epidemiology data.


Subjective and objective measures of parent-child relationship dysfunction, child separation distress, and joint attention.
The literature suggests an adverse impact of maternal stress related to interpersonal violence on parent-child interaction. The current study investigated associations between a mother's self-reported parent-child relationship dysfunction and what she does in response to her child's cues. It also examined whether maternal perception of parent-child dysfunctional interaction and child behavior when stressed by separation, along with maternal behavior in response to child distress, predicted impaired joint attention (JA) during play. Participant mothers (n = 74) and their children ages 12-48 months were recruited from community pediatrics clinics and completed two videotaped visits. After correlations, multiple linear regression was applied to find the best model fit that would predict outcomes of interest. We found that both maternal subjective report of self-reported parent-child relationship dysfunction and observed child separation distress together predicted atypical maternal behavior. Self-reported parent-child relationship dysfunction, observed atypical maternal behavior, and child separation distress combined significantly predicted less time spent in joint attention during play. Maternal posttraumatic stress predicted less maternal availability after separation stress. Clinicians should thus carefully assess and listen to parents' communication of disturbances in their relationship with their young child. Left untreated, parent-child relationship dysfunction may well lead to impairment in learning and social-emotional development.


Cell Wall Recycling-Linked Coregulation of AmpC and PenB β-Lactamases through ampD Mutations in Burkholderia cenocepacia.
In many Gram-negative pathogens, mutations in the key cell wall-recycling enzyme AmpD (N-acetyl-anhydromuramyl-L-alanine amidase) affect the activity of the regulator AmpR, which leads to the expression of AmpC β-lactamase, conferring resistance to expanded-spectrum cephalosporin antibiotics. Burkholderia cepacia complex (Bcc) species also have these Amp homologs; however, the regulatory circuitry and the nature of causal ampD mutations remain to be explored. A total of 92 ampD mutants were obtained, representing four types of mutations: single nucleotide substitution (causing an amino acid substitution or antitermination of the enzyme), duplication, deletion, and IS element insertion. Duplication, which can go through reversion, was the most frequent type. Intriguingly, mutations in ampD led to the induction of two β-lactamases, AmpC and PenB. Coregulation of AmpC and PenB in B. cenocepacia, and likely also in many Bcc species with the same gene organization, poses a serious threat to human health. This resistance mechanism is of evolutionary optimization in that ampD is highly prone to mutations allowing rapid response to antibiotic challenge, and many of the mutations are reversible in order to resume cell wall recycling when the antibiotic challenge is relieved.


Satisfaction with life and food-related life in Ecuadorian older adults.
To distinguish typologies of seniors according to their levels of satisfaction with life and food-related life, and to characterize them according to the importance assigned to family, health-related aspects, eating habits, nutritional status and sociodemographic characteristics. A survey was administered to a non-probabilistic sample of 817 seniors aged 60 years and over, of both genders, resident in urban and rural zones of the province of Guayas, Ecuador. The instrument included: Satisfaction with Life Scale (SWLS), Satisfaction with Food-related Life (SWFL), the Health-related Quality of Life Index (HRQOL-4) and the Family Importance Scale (FIS). Eating habits, sociodemographic variables, approximate weight and height were asked about. A cluster analysis was used to distinguish three typologies that differed significantly in the scores on the SWLS, SWFL, FIS, number of days with mental health problems, self-perception of health status, body mass index, eating habits and zone of residence. Two typologies presented high levels of satisfaction with life and food-related life, although the variables associated with overall well-being and in the food domain were different in each typology. The results suggest that high levels of satisfaction with life and food-related life in Ecuadorian seniors are associated with both healthful eating habits and having lunch in company, fewer mental health problems and greater importance assigned to family.


Variations in the G6PC2/ABCB11 genomic region are associated with fasting glucose levels.
Identifying the genetic variants that regulate fasting glucose concentrations may further our understanding of the pathogenesis of diabetes. We therefore investigated the association of fasting glucose levels with SNPs in 2 genome-wide scans including a total of 5,088 nondiabetic individuals from Finland and Sardinia. We found a significant association between the SNP rs563694 and fasting glucose concentrations (P = 3.5 x 10(-7)). This association was further investigated in an additional 18,436 nondiabetic individuals of mixed European descent from 7 different studies. The combined P value for association in these follow-up samples was 6.9 x 10(-26), and combining results from all studies resulted in an overall P value for association of 6.4 x 10(-33). Across these studies, fasting glucose concentrations increased 0.01-0.16 mM with each copy of the major allele, accounting for approximately 1% of the total variation in fasting glucose. The rs563694 SNP is located between the genes glucose-6-phosphatase catalytic subunit 2 (G6PC2) and ATP-binding cassette, subfamily B (MDR/TAP), member 11 (ABCB11). Our results in combination with data reported in the literature suggest that G6PC2, a glucose-6-phosphatase almost exclusively expressed in pancreatic islet cells, may underlie variation in fasting glucose, though it is possible that ABCB11, which is expressed primarily in liver, may also contribute to such variation.


Lymphotoxin acts as an autocrine growth factor for Epstein-Barr virus-transformed B cells and differentiated Burkitt lymphoma cell lines.
A critical event in B cell immortalization by Epstein-Barr virus (EBV) is the establishment of an autocrine loop where cells produce a growth factor which supports their own proliferation. We investigated the potential of lymphoblastoid cell lines (LCL) and Burkitt lymphoma (BL) cell lines to produce and respond to the cytotoxins, tumor necrosis factor-alpha (TNF-alpha) and lymphotoxin (LT). Transformation in vitro of peripheral blood B cells by EBV from seven different donors resulted in spontaneous production of both LT (11,542 pg/ml +/- 7546, mean +/- SD) and, to a lesser extent, TNF-alpha (197 pg/ml +/- 174). Similarly BL cell lines derived from in vivo transformation which developed a 'LCL-like' phenotype in vitro (group III) produced more LT (1990 pg/ml +/- 1740) than the 'group I' BL cell lines (< 40 pg/ml LT) which had maintained the original BL biopsy cell phenotype in vitro. Transformation of peripheral blood B cells to generate LCL also resulted in an increase in surface p75 (p < 0.02) and to a lesser extent p55 (not significant, ns) TNF receptor (TNF-R) expression. Similar increases in surface TNF-R (p75 p < 0.02, p55 ns) were observed on the 'group III' BL cell lines compared with the 'group I' BL cell lines. Proliferation of an LCL and a 'group III' BL cell line in vitro was via an autocrine loop since inhibition of LT reduced proliferation. This proliferation could also be blocked in the presence of the antagonistic anti-p55 TNF-R antibody, H398, but not the antagonistic antibody anti-p75 TNF-R antibody UTR-1. Furthermore, proliferation could be induced with the p55 agonistic antibody, HTR-9. In contrast to these observations with p55 TNF-R antibodies, two out of six of the 'group III' BL lines (Jijoye and Oba) only expressed the p75 TNF-R and proliferation of these cells could only be blocked by the antagonistic anti-p75 TNF-R antibody UTR-1. These data suggest that LT is an autocrine growth factor for lymphoblastoid cells, and BL cell lines which display an LCL phenotype. Furthermore, although both TNF-R are increased on the surface of these cells, this autocrine growth signal is mediated principally through binding to the p55 TNF-R.


Completing Baseline Mapping of Trachoma in Uganda: Results of 14 Population-Based Prevalence Surveys Conducted in 2014 and 2018.
We aimed to estimate the prevalence of trachomatous inflammation-follicular (TF) in children aged 1-9 years, trichiasis in adults aged ≥15 years, and water and sanitation (WASH) indicators in 12 suspected-endemic districts in Uganda. Surveys were undertaken in 14 evaluation units (EUs) covering 12 districts. Districts were selected based on a desk review in 2014 (four districts) and trachoma rapid assessments in 2018 (eight districts). We calculated that 1,019 children aged 1-9 years were needed in each EU to estimate TF prevalence with acceptable precision and used three-stage cluster sampling to select 30 households in each of 28 (2014 surveys) or 24 (2018 surveys) villages. Participants living in selected households aged ≥1 year were examined for trachoma; thus enabling estimation of prevalences of TF in 1-9 year-olds and trichiasis in ≥15 year-olds. Household-level WASH access data were also collected. A total of 11,796 households were surveyed; 22,465 children aged 1-9 years and 24,652 people aged ≥15 years were examined. EU-level prevalence of TF ranged from 0.3% (95% confidence interval [CI] 0.1-0.7) to 3.9% (95% CI 2.1-5.8). EU-level trichiasis prevalence ranged from 0.01% (95% CI 0-0.11) to 0.81% (95% CI 0.35-1.50). Overall proportions of households with improved drinking water source, water source in yard or within 1km, and improved sanitation facilities were 88.1%, 23.0% and 23.9%, respectively. TF was not a public health problem in any of the 14 EUs surveyed: antibiotic mass drug administration is not required in these districts. However, in four EUs, trichiasis prevalence was ≥ 0.2%, so public health-level trichiasis surgery interventions are warranted. These findings will facilitate planning for elimination of trachoma in Uganda.


Treponema pallidum and Borrelia burgdorferi lipoproteins and synthetic lipopeptides activate monocytes/macrophages.
The observation that the major membrane immunogens of the spirochetal pathogens. Treponema pallidum and Borrelia burgdorferi are lipoproteins prompted studies to investigate macrophage activation by the 47-kDa lipoprotein of T. pallidum and the acylated outer surface protein A (OspA) of B. burgdorferi. Both lipoproteins induced the synthesis of biologically active TNF-alpha and chloramphenicol acetyltransferase in a murine macrophage cell line transfected with a chloramphenicol acetyltransferase reporter gene controlled by a TNF promoter (TB2 cells). Nonacylated forms of these polypeptides did not induce cell activation. Comparison between purified OspA and B. burgdorferi cellular lipids revealed that the former was the more potent inducer of TNF-alpha. Synthetic lipohexapeptides corresponding to the N-termini of the 47-kDa lipoprotein of T. pallidum and OspA also activated TB2 cells in a dose-dependent fashion, whereas the nonlipidated hexapeptides were without effect, further underscoring the importance of protein acylation to cell activation. Among several lines of evidence supporting that macrophage stimulation by LPS and lipopeptides proceeds via different mechanisms, the most notable was that lipopeptides activated peritoneal macrophages from LPS-nonresponsive C3H/HeJ mice. The potential for spirochetal lipoproteins to function as general macrophage activators was demonstrated by the ability of the synthetic analogues to induce IL-1 beta, IL-6, and IL-12, in addition to TNF, in murine and/or human macrophages. Our findings indicate that spirochetal lipoproteins may be important immunomodulators in syphilis and Lyme disease and that the synthetic lipopeptides will be useful surrogates for studying immune mechanisms operative in the two spirochetal diseases.


Psychological Correlates of Sedentary Screen Time Behaviour Among Children and Adolescents: a Narrative Review.
The aims of this narrative review were to (1) synthesise the literature on the relationship between screen time and important mental health outcomes and (2) examine the underpinning factors that can influence this association. Paralleling the rise of mental health issues in children and adolescents is the ubiquitous overuse of screens, but it is unclear how screen time is related to important mental health outcomes and whether this association differs by gender, age and screen type. Medline/PubMed, PsychINFO and Google Scholar databases were searched on December 2019 for articles published mainly in the last 5 years. The search focused on two main concepts: (i) screen time and (ii) mental health outcomes including anxiety, depression, psychological and psychosocial well-being and body image concerns. Sixty studies were included in the review. Higher levels of screen time were associated with more severe depressive symptoms. We found moderate evidence for an association between screen time and poor psychological well-being and body dissatisfaction especially among females. Relationships between screen time and anxiety were inconsistent and somewhat gender specific. Social media use was consistently associated with poorer mental health. Higher levels of screen time are generally associated with poorer mental health outcomes, but associations are influenced by screen type, gender and age. Practitioners, parents, policy makers and researchers should collectively identify and evaluate strategies to reduce screen time, or to use screens more adaptively, as a means of promoting better mental health among children and adolescents.


Beneficial effects of fenugreek glycoside supplementation in male subjects during resistance training: A randomized controlled pilot study.
To evaluate the efficacy and safety of the glycoside fraction of fenugreek (Trigonella foenum-graecum) seeds (Fenu-FG) on physiological parameters related to muscle anabolism, androgenic hormones, and body fat in healthy male subjects during an 8-week resistance training program using a prospective, randomized, double-blind, placebo controlled design. Sixty healthy male subjects were randomized to ingest capsules of Fenu-FG (1 capsule of 300 mg, twice per day) or the matching placebo at a 1:1 ratio. The subjects participated in a supervised 4-day per week resistance-training program for 8 weeks. The outcome measurements were recorded at recruitment (baseline) and at the end of the treatment (8 weeks). The efficacy outcome included serum testosterone (total and free) levels, muscle strength and repetitions to failure, metabolic markers for anabolic activity (serum creatinine and blood urea nitrogen), and % body fat. The standard safety measurements such as adverse events monitoring, vital signs, hematology, biochemistry, and urinalysis were performed. Fenu-FG supplementation demonstrated significant anabolic and androgenic activity as compared with the placebo. Fenu-FG treated subjects showed significant improvements in body fat without a reduction in muscle strength or repetitions to failure. The Fenu-FG supplementation was found to be safe and well-tolerated. Fenu-FG supplementation showed beneficial effects in male subjects during resistance training without any clinical side effects.


Causes of death contributing to changes in life expectancy: United States, 1984-89.
During 1984-89, life expectancy increased for the total population (0.6 year), white males (0.9 year), and white females (0.5 year), while it decreased for black males (-0.8year) and black females (-0.2 year). These changes in life expectancy are the result of changes in mortality for specific age-ram-sex groups and identifiable causes of death. Using a technique developed by Eduardo E. Arriaga (1,2), this report identifies major causes of death contributing to the change in life expectancy between 1984 and 1989. Refer to Technical notes for a brief description of this methodology. The tectique partitions changes in life expectancy into positive contributions (which are those causes of death and age groups that contribute to an increase in life expectancy) and negative contributions (which are those that contribute to a decrease). The report includes separate analyses by age and causes of death. For the total population, positive contributions occurred for the age groups under 15 years and age groups 45 years and over while negative contributions occurred for the age groups 15-44 years. The report also examines the combined contributions of age and causes of death for the total population and the four race-sex groups. For the total population, positive contributions to the increase in life expectancy were due to changes in mortality for Diseases of heart under 1 and 35 years and over; Cerebrovascular diseases, 40 years and over; and Malignant neoplasms, including neoplasms of lymphatic and hematopoietic tissues, 30-59 years. Negative contributions were largely due to HIV infection, 20-59 years; Malignant neoplasma, including neoplasm of lymphatic and hematopoietic tissues, 65 years and oveq and Pneumonia and influenza, 75 years and over.


Activation of peripheral leukocytes in rat pregnancy and experimental preeclampsia.
The aim of this study was to search for activation markers of peripheral leukocytes in experimental preeclampsia in the rat. Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 microg/kg body weight). For comparison, rats with normal pregnancies that were infused with sodium chloride solution and cyclic rats that were infused with either endotoxin or sodium chloride solution were used. At various points before and after the infusion, blood samples were withdrawn and analyzed by means of whole-blood flow cytometry to evaluate expression of inflammation-associated adhesion molecules (CD11b, CD11a, CD49d, and CD62L) and CD14 on the leukocytes. Normal pregnancy was associated with increased CD11b (granulocytes and monocytes), CD11a (monocytes and lymphocytes), and CD49d (granulocytes, monocytes, and lymphocytes) expression. In addition to these changes found in normal pregnancy, reduced CD62L and increased CD11a and CD49d expression was found on granulocytes after endotoxin treatment of pregnant rats. No effect of endotoxin was observed in cyclic rats. Leukocytes of rats with experimental preeclampsia and, to a lesser extent, those of rats with normal pregnancies had an activated phenotype. These results are consistent with our previous findings in human subjects and suggest that (experimental) preeclampsia results from a generalized inflammatory response.


Role of phenolic O-H and methylene hydrogen on the free radical reactions and antioxidant activity of curcumin.
To understand the relative importance of phenolic O-H and the CH-H hydrogen on the antioxidant activity and the free radical reactions of Curcumin, (1,7-bis[4-hydroxy-3-methoxyphenyl]-1,6-heptadiene-3,5-dione), biochemical, physicochemical, and density functional theory (DFT) studies were carried out with curcumin and dimethoxy curcumin (1,7-bis[3, 4-dimethoxy phenyl]-1,6-heptadiene-3,5-dione). The antioxidant activity of these compounds was tested by following radiation-induced lipid peroxidation in rat liver microsomes, and the results suggested that at equal concentration, the efficiency to inhibit lipid peroxidation is changed from 82% with curcumin to 24% with dimethoxy curcumin. Kinetics of reaction of (2,2'-diphenyl-1-picrylhydrazyl) DPPH, a stable hydrogen abstracting free radical was tested with these two compounds using stopped-flow spectrometer and steady state spectrophotometer. The bimolecular rate constant for curcumin was found to be approximately 1800 times greater than that for the dimethoxy derivative. Cyclic voltammetry studies of these two systems indicated two closely lying oxidation peaks at 0.84 and 1.0 V vs. SCE for curcumin, while only one peak at 1.0 V vs. SCE was observed for dimethoxy curcumin. Pulse radiolysis induced one-electron oxidation of curcumin and dimethoxy curcumin was studied at neutral pH using (*)N(3) radicals. This reaction with curcumin produced phenoxyl radicals absorbing at 500 nm, while in the case of dimethoxy curcumin a very weak signal in the UV region was observed. These results suggest that, although the energetics to remove hydrogen from both phenolic OH and the CH(2) group of the beta-diketo structure are very close, the phenolic OH is essential for both antioxidant activity and free radical kinetics. This is further confirmed by DFT calculations where it is shown that the -OH hydrogen is more labile for abstraction compared to the -CH(2) hydrogen in curcumin. Based on various experimental and theoretical results it is definitely concluded that the phenolic OH plays a major role in the activity of curcumin.


Detecting and avoiding likely false-positive findings - a practical guide.
Recently there has been a growing concern that many published research findings do not hold up in attempts to replicate them. We argue that this problem may originate from a culture of 'you can publish if you found a significant effect'. This culture creates a systematic bias against the null hypothesis which renders meta-analyses questionable and may even lead to a situation where hypotheses become difficult to falsify. In order to pinpoint the sources of error and possible solutions, we review current scientific practices with regard to their effect on the probability of drawing a false-positive conclusion. We explain why the proportion of published false-positive findings is expected to increase with (i) decreasing sample size, (ii) increasing pursuit of novelty, (iii) various forms of multiple testing and researcher flexibility, and (iv) incorrect P-values, especially due to unaccounted pseudoreplication, i.e. the non-independence of data points (clustered data). We provide examples showing how statistical pitfalls and psychological traps lead to conclusions that are biased and unreliable, and we show how these mistakes can be avoided. Ultimately, we hope to contribute to a culture of 'you can publish if your study is rigorous'. To this end, we highlight promising strategies towards making science more objective. Specifically, we enthusiastically encourage scientists to preregister their studies (including a priori hypotheses and complete analysis plans), to blind observers to treatment groups during data collection and analysis, and unconditionally to report all results. Also, we advocate reallocating some efforts away from seeking novelty and discovery and towards replicating important research findings of one's own and of others for the benefit of the scientific community as a whole. We believe these efforts will be aided by a shift in evaluation criteria away from the current system which values metrics of 'impact' almost exclusively and towards a system which explicitly values indices of scientific rigour.


Ruptured saccular aneurysm arising from fenestrated proximal anterior cerebral artery : case report and literature review.
The aneurysm arising from fenestrated proximal anterior cerebral artery (ACA) is considered to be unique. The authors report a case of a 59-year-old woman who presented with a subarachnoid hemorrhage (SAH) secondary to a ruptured aneurysm originating from the fenestrated A1 segment of right ACA. The patient had another unruptured aneurysm which was located at the right middle cerebral artery bifurcation. She was successfully treated with surgical clipping for both aneurysms. From the previously existing literatures, we found 18 more cases (1983-2011) of aneurysms associated with fenestrated A1 segment. All cases represented saccular type of aneurysms, and 79% of the patients had SAH. There were three subtypes of the fenestrated A1 aneurysms depending on the anatomical location, relative to the fenestrated segment. The most common type was the aneurysms located on the proximal end of fenestrated artery (82%). Azygos ACA and hypoplastic A1 were frequently accompanied by the aneurysm (33% and 31%, respectively), and multiple aneurysms were shown in three cases (16%). Considering that fenestrated A1 segment is likely to develop an aneurysm, which has high risk of rupture, early management may benefit patients with aneurysms accompanied by fenestrated proximal ACA.


[Carcinoembryonic antigen (CEA) as an independent prognostic factor in colorectal carcinoma].
CEA is widely used in the follow up of patients with colorectal carcinoma. To study the value of preoperative CEA as an independent prognostic factor in colorectal carcinoma. Analysis of 373 operated patients (204 females, age range 21-92 years) with colorectal carcinoma and a mean follow up of 53 months. The cutoff value for CEA was 5 ng/ml. Ninety four percent of patients had an excisable tumor, 79% had involvement of perirectal/pericolonic adipose tissue and 46% had lymph node involvement. Staging was done using Dukes-Turnbull and TNM classifications. CEA was normal in 61% of cases, over 5 ng/ml in 39% and over 15 ng/ml in 22%. There was a strong correlation between mean preoperative CEA and tumor stage, depth and lymph node involvement. During the follow up, 140 patients died, 57 with normal and 83 with elevated CEA. Cancer mortality in patients subjected to a curative excision of the tumor (Dukes A-C2/TNM I-III) was 9% for colonic tumors and 36% for rectal tumors (p < 0.001). There were no survival differences in patients with Dukes B/TNM II tumors according to preoperative CEA. Among Dukes C/TNM III tumors, survival difference was only significant for rectal tumors. A Cox model disclosed tumor stage, location and preoperative CEA as independent prognostic factors for survival. CEA is an independent prognostic factor for survival in colorectal carcinoma and high levels suggest an advanced disease.


Sphenoidal fontanelle area measurement on rendered three-dimensional ultrasound.
Rendered three-dimensional images of the fetal cranial vault permit the identification of sutures and measurement of the fontanelle surface area. The objective of this study was to establish the feasibility and repeatability of sphenoidal fontanelle area measurement. Head volumes were obtained using the biparietal diameter as the reference plane. These were displayed with maximum rendering and the surface area of the sphenoidal fontanelle was measured by one operator. Feasibility was expressed as the percentage of scans with a successful measurement at different gestational ages. Repeatability was expressed as coefficient of variation (CV), intraclass correlation coefficient (ICC) and Bland-Altman plots. Thirty women underwent 141 scans. Feasibility of sphenoidal fontanelle measurement is over 90% before 26 weeks, but drops gradually thereafter and becomes 0% at 34 weeks. Intraobserver repeatability of measurement on the same scan image demonstrated CV = 17.5% and ICC = 0.883; however, when measuring a newly acquired volume, the repeatability is poor, CV = 30.7% and ICC = 0.649. Although it is feasible to visualize the sphenoidal fontanelle on rendered three dimensions, its surface area measurement is not reliable for clinical use. There is potential to overestimate repeatability when remeasuring the same volume.


Novel targets for hepatitis B virus therapy.
Treatment with either pegylated interferon-alpha (pegIFN-α) or last generation nucleos(t)ide analogues (NAs) successfully leads to serum viral load suppression in most chronically infected hepatitis B (CHB) patients, but HBsAg loss is only achieved in 10% of the cases after a 5-year follow-up. Thus, therapy must be administered long-term and it will not completely eliminate infection because of the persistent hepatitis B virus (HBV) minichromosome in infected cells, and cannot completely abolish the risk of developing severe sequelae such as cirrhosis and hepatocellular carcinoma. Recent progress in the development of in vitro and in vivo models of HBV infection have helped renew interest in the investigation of the viral life cycle, as well as specific virus-host cell interactions to identify new targets for the development of new antiviral drugs. This includes either direct inhibition of viral replication by targeting fundamental steps such as entry, cccDNA formation/stability, viral transcripts, capsid assembly and secretion or the manipulation of the host immune system for better defence against infection. Multiple strategies are currently under investigation, including boosting endogenous innate responses and/or restoring adaptive immunity via engineering of HBV-specific T cells or via the use of inhibitors of negative regulators, as well as therapeutic vaccines. It is increasingly clear that multiple therapeutic strategies must be combined to reach a cure of HBV and that the definition of clinical, virological and immunological correlates for the management of treatment are urgently needed.


Plant species radiations: where, when, why?
The spatial and temporal patterns of plant species radiations are largely unknown. I used a nonlinear regression to estimate speciation and extinction rates from all relevant dated clades. Both are surprisingly high. A high species richness can be the result of either little extinction, thus preserving the diversity that dates from older radiations (a 'mature radiation'), or a 'recent and rapid radiation'. The analysis of radiations from different regions (Andes, New Zealand, Australia, southwest Africa, tropics and Eurasia) revealed that the diversity of Australia may be largely the result of mature radiations. This is in sharp contrast to New Zealand, where the flora appears to be largely the result of recent and rapid radiations. Mature radiations are characteristic of regions that have been climatically and geologically stable throughout the Neogene, whereas recent and rapid radiations are more typical of younger (Pliocene) environments. The hyperdiverse Cape and Neotropical floras are the result of the combinations of mature as well as recent and rapid radiations. Both the areas contain stable environments (the Amazon basin and the Cape Fold Mountains) as well as dynamic landscapes (the Andes and the South African west coast). The evolution of diversity can only be understood in the context of the local environment.


Interspecific and intraspecific variation in cercariae release.
Given the importance of temperature for the shedding of trematode infective stages (cercariae) from gastropod first intermediate hosts, we investigated the response to temperature increases of trematodes of the intertidal snail Zeacumantus subcarinatus collected from different latitudes (differing in mean annual summer temperature) on the South Island of New Zealand. We investigated whether shedding of cercariae at elevated temperatures differed both between species (interspecific variation) and among populations of each trematode species (intraspecific variation). The shedding of one trematode species, Maritrema novaezealandensis (Microphallidae), appears to decrease with increased temperature and differed among locations. In contrast, the shedding of a second species, Acanthoparyphium sp. (Echinostomatidae), increased at warmer temperatures. In particular, Acanthoparyphium sp.-infected snails from 1 location showed the greatest increase in the shedding of cercariae in response to elevated temperature. Our results demonstrate that different trematode species and populations may be dissimilarly impacted by temperature changes resulting from global warming. In addition, both interspecific and intraspecific variation may result in different impacts of trematodes on ecosystems at different locales.


Analysis of the meal-dependent intragastric performance of a gastric-retentive tablet assessed by magnetic resonance imaging.
Modern medical imaging modalities can trace labelled oral drug dosage forms in the gastrointestinal tract, and thus represent important tools for the evaluation of their in vivo performance. The application of gastric-retentive drug delivery systems to improve bioavailability and to avoid unwanted plasma peak concentrations of orally administered drugs is of special interest in clinical and pharmaceutical research. To determine the influence of meal composition and timing of tablet administration on the intragastric performance of a gastric-retentive floating tablet using magnetic resonance imaging in the sitting position. A tablet formulation was labelled with iron oxide particles as negative magnetic resonance contrast marker to allow the monitoring of the tablet position in the food-filled human stomach. Labelled tablet was administered, together with three different solid meals, to volunteers seated in a 0.5-T open-configuration magnetic resonance system. Volunteers were followed over a 4-h period. Labelled tablet was detectable in all subjects throughout the entire study. The tablet showed persistent good intragastric floating performance independent of meal composition. Unfavourable timing of tablet administration had a minor effect on the intragastric tablet residence time and floating performance. Magnetic resonance imaging can reliably monitor and analyse the in vivo performance of labelled gastric-retentive tablets in the human stomach.


[Roentgeno-endovascular surgery of hypofunctional ovaries in varicosities of the ovarian veins].
Three degrees of ovarian varicocele in sterile women or women with menstrual disturbance were described on the basis of analysis of 62 superselective ovariophlebograms. A nonsurgical pathogenetic method for correction of ovarian hypofunction in disturbed circulation in the venous region of the pelvic organs was developed by cutting off the ovarian vein from the pathological renocaval vascular shunt with the help of roentgenovascular intervention. It is based upon transcatheter occlusion of the left ovarian vein and directly follows diagnostic phlebography. Roentgenoendovascular occlusion of the left ovarian vein was performed in 41 patients Ivanissevich's operation--in 8 patients. In 4-18 months after intervention, improved results of a clinical picture and functional diagnostic tests were observed in 46 patients. Of 19 women with passable uterine tubes 14 got pregnant.


Intravenous micronutrient therapy (Myers' Cocktail) for fibromyalgia: a placebo-controlled pilot study.
Intravenous micronutrient therapy (IVMT), and specifically the Myers' Cocktail, is a popular approach for treating fibromyalgia syndrome (FMS) among complementary and alternative medicine practitioners, but its efficacy is uncertain. This trial assessed the feasibility, safety, and provided insights into the efficacy of this therapy. This was a randomized, double-blind, placebo-controlled pilot study. The study locations were an academic research center, teaching hospital, and affiliated Integrative Medicine Center in Derby, CT. The subjects were 34 adults with American College of Rheumatology (ACR)-defined FMS. Subjects were randomly assigned either to treatment (weekly infusions of IVMT) or to placebo (weekly infusions of lactated Ringer's solution) for 8 weeks. Primary outcome was change in the Tender Point Index, assessed 8 and 12 weeks after initiation. Secondary measures included a Visual Analog Scale to assess global pain, and validated measures of physical function (Fibromyalgia Impact Questionnaire), mood (Beck Depression Index), and quality of life (Health Status Questionnaire 2.0). Clinically significant improvements were noted (of a magnitude similar to other effective interventions). However, in part because of the high placebo response and the small sample size, no statistically significant differences were seen between groups, in any outcome measure, at 8 and 16 weeks. Statistically significant within-group differences were seen in both the intervention and placebo groups, demonstrating a treatment effect for both IVMT and placebo. At 8 weeks, the IVMT group experienced significantly improved tender points, pain, depression, and quality of life directly following treatment (all p < or = 0.02), while the placebo group experienced significantly improved tender points only (p < or = 0.05). The treatment effects of IVMT persisted at 4 weeks postintervention for tender points, pain, and quality of life, while placebo effects persisted only for tender points. A single minor adverse event was noted in one subject in the intervention group. This first controlled pilot study established the safety and feasibility of treating FMS with IVMT. Most subjects experienced relief as compared to baseline, but no statistically significant differences were seen between IVMT and placebo. The efficacy of IVMT for fibromyalgia, relative to placebo, is as yet uncertain.


Role of non-specific DNA in reducing coding DNA requirement for transient gene expression with CHO and HEK-293E cells.
Transient gene expression (TGE) is a rapid method for the production of recombinant proteins in mammalian cells. While the TGE volumetric productivity has improved significantly over the past decade, the amount of plasmid DNA (pDNA) needed for transfection remains very high. Here, we examined the use of non-specific (filler) DNA to partially replace the transgene-bearing plasmid DNA (coding pDNA) in transfections of Chinese hamster ovary (CHO) and human embryo kidney (HEK-293E) cells. When the optimal amount of coding pDNA for either host was reduced by 67% and replaced with filler DNA, the recombinant protein yield decreased by only 25% relative to the yield in control transfections. Filler DNA did not affect the cellular uptake or intracellular stability of coding pDNA, but its presence lead to increases of the percentage of transfected cells and the steady-state level of transgene mRNA compared to control transfections. Studies of the physicochemical properties of DNA-polyethyleneimine (PEI) complexes with or without filler DNA did not reveal any differences in their size or surface charge. The results suggest that filler DNA allows the coding pDNA to be distributed over a greater number of DNA-PEI complexes, leading to a higher percentage of transfected cells. The co-assembly of filler DNA and coding pDNA within complexes may also allow the latter to be more efficiently utilized by the cell's transcription machinery, resulting in a higher level of transgene mRNA.


Functional polymorphism in ABCA1 influences age of symptom onset in coronary artery disease patients.
ATP-binding-cassette-transporter-A1 (ABCA1) plays a pivotal role in intracellular cholesterol removal, exerting a protective effect against atherosclerosis. ABCA1 gene severe mutations underlie Tangier disease, a rare Mendelian disorder that can lead to premature coronary artery disease (CAD), with age of CAD onset being two decades earlier in mutant homozygotes and one decade earlier in heterozygotes than in mutation non-carriers. It is unknown whether common polymorphisms in ABCA1 could influence age of symptom onset of CAD in the general population. We examined common promoter and non-synonymous coding polymorphisms in relation to age of symptom onset in a group of CAD patients (n = 1164), and also carried out in vitro assays to test effects of the promoter variations on ABCA1 promoter transcriptional activity and effects of the coding variations on ABCA1 function in mediating cellular cholesterol efflux. Age of symptom onset was found to be associated with the promoter - 407G > C polymorphism, being 2.82 years higher in C allele homozygotes than in G allele homozygotes and intermediate in heterozygotes (61.54, 59.79 and 58.72 years, respectively; P = 0.002). In agreement, patients carrying ABCA1 haplotypes containing the -407C allele had higher age of symptom onset. Patients of the G/G or G/C genotype of the -407G > C polymorphism had significant coronary artery stenosis (>75%) at a younger age than those of the C/C genotype (P = 0.003). Reporter gene assays showed that ABCA1 haplotypes bearing the -407C allele had higher promoter activity than haplotypes with the -407G allele. Functional analyses of the coding polymorphisms showed an effect of the V825I substitution on ABCA1 function, with the 825I variant having higher activity in mediating cholesterol efflux than the wild-type (825V). A trend towards higher symptom onset age in 825I allele carriers was observed. The data indicate an influence of common ABCA1 functional polymorphisms on age of symptom onset in CAD patients.


Platelet endothelial cell adhesion molecule-1 mediates endothelial-cardiomyocyte communication and regulates cardiac function.
Dilated cardiomyopathy is characterized by impaired contractility of cardiomyocytes, ventricular chamber dilatation, and systolic dysfunction. Although mutations in genes expressed in the cardiomyocyte are the best described causes of reduced contractility, the importance of endothelial-cardiomyocyte communication for proper cardiac function is increasingly appreciated. In the present study, we investigate the role of the endothelial adhesion molecule platelet endothelial cell adhesion molecule (PECAM-1) in the regulation of cardiac function. Using cell culture and animal models, we show that PECAM-1 expressed in endothelial cells (ECs) regulates cardiomyocyte contractility and cardiac function via the neuregulin-ErbB signaling pathway. Conscious echocardiography revealed left ventricular (LV) chamber dilation and systolic dysfunction in PECAM-1(-/-) mice in the absence of histological abnormalities or defects in cardiac capillary density. Despite deficits in global cardiac function, cardiomyocytes isolated from PECAM-1(-/-) hearts displayed normal baseline and isoproterenol-stimulated contractility. Mechanistically, absence of PECAM-1 resulted in elevated NO/ROS signaling and NRG-1 release from ECs, which resulted in augmented phosphorylation of its receptor ErbB2. Treatment of cardiomyocytes with conditioned media from PECAM-1(-/-) ECs resulted in enhanced ErbB2 activation, which was normalized by pre-treatment with an NRG-1 blocking antibody. To determine whether normalization of increased NRG-1 levels could correct cardiac function, PECAM-1(-/-) mice were treated with the NRG-1 blocking antibody. Echocardiography showed that treatment significantly improved cardiac function of PECAM-1(-/-) mice, as revealed by increased ejection fraction and fractional shortening. We identify a novel role for PECAM-1 in regulating cardiac function via a paracrine NRG1-ErbB pathway. These data highlight the importance of tightly regulated cellular communication for proper cardiac function.


New histone supply regulates replication fork speed and PCNA unloading.
Correct duplication of DNA sequence and its organization into chromatin is central to genome function and stability. However, it remains unclear how cells coordinate DNA synthesis with provision of new histones for chromatin assembly to ensure chromosomal stability. In this paper, we show that replication fork speed is dependent on new histone supply and efficient nucleosome assembly. Inhibition of canonical histone biosynthesis impaired replication fork progression and reduced nucleosome occupancy on newly synthesized DNA. Replication forks initially remained stable without activation of conventional checkpoints, although prolonged histone deficiency generated DNA damage. PCNA accumulated on newly synthesized DNA in cells lacking new histones, possibly to maintain opportunity for CAF-1 recruitment and nucleosome assembly. Consistent with this, in vitro and in vivo analysis showed that PCNA unloading is delayed in the absence of nucleosome assembly. We propose that coupling of fork speed and PCNA unloading to nucleosome assembly provides a simple mechanism to adjust DNA replication and maintain chromatin integrity during transient histone shortage.


Worsening or development of syringomyelia following Chiari I decompression: case report.
The effects of posterior fossa decompression on Chiari malformation Type I-induced syringomyelia have been well described. However, treatment of worsening syringomyelia after Chiari decompression remains enigmatic. This paper defines patient and clinical characteristics as well as treatment and postoperative radiological and clinical outcomes in patients experiencing this complication. The authors performed a retrospective review of patients at the Children's Hospital of Pittsburgh and Children's of Alabama who developed worsening syringomyelia after Chiari decompression was performed. Fourteen children (age range 8 months to 15 years), 7 of whom had preoperative syringomyelia, underwent posterior fossa decompression. Aseptic meningitis (n = 3) and bacterial meningitis (n = 2) complicated 5 cases (4 of these patients were originally treated at outside hospitals). Worsening syringomyelia presented a median of 1.4 years (range 0.2-10.3 years) after the primary decompression. Ten children presented with new, recurrent, or persistent symptoms, and 4 were asymptomatic. Secondary Chiari decompression was performed in 11 of the 14 children. The other 3 children were advised to undergo secondary decompression. A structural cause for each failed primary Chiari decompression (for example, extensive scarring, suture in the obex, arachnoid web, residual posterior arch of C-1, and no duraplasty) was identified at the secondary operation. After secondary decompression, 8 patients' symptoms completely resolved, 1 patient's condition stabilized, and 2 patients remained asymptomatic. Radiologically, 10 of the 11 children had a decrease in the size of their syringes, and 1 child experienced no change (but improved clinically). The median follow-up from initial Chiari decompression was 3.1 years (range 0.8-14.1 years) and from secondary decompression, 1.3 years (range 0.3-4.5 years). No patient underwent syringopleural shunting or other nonposterior fossa treatment for syringomyelia. Based on the authors' experience, children with worsening syringomyelia after decompression for Chiari malformation Type I generally have a surgically remediable structural etiology, and secondary exploration and decompression should be considered.


Identification of virulence determinants of the human pathogenic fungi Aspergillus fumigatus and Candida albicans by proteomics.
Both fungi Candida albicans and Aspergillus fumigatus can cause a number of life-threatening systemic infections in humans. The commensal yeast C. albicans is one of the main causes of nosocomial fungal infectious diseases, whereas the filamentous fungus A. fumigatus has become one of the most prevalent airborne fungal pathogens. Early diagnosis of these fungal infections is challenging, only a limited number of antifungals for treatment are available, and the molecular details of pathogenicity are hardly understood. The completion of both the A. fumigatus and C. albicans genome sequence provides the opportunity to improve diagnosis, to define new drug targets, to understand the functions of many uncharacterised proteins, and to study protein regulation on a global scale. With the application of proteomic tools, particularly two-dimensional gel electrophoresis and LC/MS-based methods, a comprehensive overview about the proteins of A. fumigatus and C. albicans present or induced during environmental changes and stress conditions has been obtained in the past 5 years. However, for the discovery of further putative virulence determinants, more sensitive and targeted proteomic methods have to be applied. Here, we review the recent proteome data generated for A. fumigatus and C. albicans that are related to factors required for pathogenicity.


Phase II study of direct intralesional gene transfer of allovectin-7, an HLA-B7/beta2-microglobulin DNA-liposome complex, in patients with metastatic melanoma.
Cutaneous melanoma is one of the most rapidly increasing cancers in the United States. Because of the lack of effective treatment options and toxicities of most chemotherapeutic and radiation regimes, immunotherapies such as vaccination therapy represent an attractive approach for patients with advanced melanoma. The purpose of this study was to evaluate the response rate, time to progression, and survival of patients with metastatic melanoma treated by direct intratumoral injection with Allovectin-7 (a plasmid DNA encoding the genes HLA-B7 and beta2-microglobulin complexed with a cationic lipid mixture, DMRIE/DOPE. Fifty-two patients with metastatic melanoma were enrolled in this Phase II study. Therapy consisted of six intratumoral injections of 10 microg of Allovectin-7 over a 9-week period. Treatment was well tolerated. Treatment-related adverse events were mild to moderate, the most frequent of which were ecchymosis, pruritus (and/or discomfort at the injection site), and pneumothoraces. Regression of the injected lesion was observed in 18% of patients, including one complete response, three partial responses, and five minor responses. An overall response rate of 4% (two partial responses) was documented, and nine patients (18%) maintained stable disease for at least 11 weeks. Six patients remained alive 25.1 to 39.4 months from their first injection, including two patients with local (injected tumor) responses and one patient with an overall disease partial response. This study demonstrates that intratumoral administration of Allovectin-7 in metastatic melanoma is safe and can produce both responses in injected lesions and in overall disease. Clinical trials optimizing patient selection and combining Allovectin-7 with other modalities of therapy are currently ongoing in an effort to improve response rates.


Long-term sequelae after acquired pediatric hemorrhagic cerebellar lesions.
The aim of the present study was to assess cognitive, affective, and motor long-term sequelae after acquired focal pediatric cerebellar lesions. Eight patients with a history of isolated acquired hemorrhagic cerebellar lesions before the age of 13 participated in this study. All participants underwent a neurologic examination, including the Zurich Neuromotor Assessment (ZNA) and the International Cooperative Ataxia Rating Scale (ICARS). Cognitive functions have been evaluated with a general cognitive assessment and an extensive neuropsychological battery. Furthermore, patients and parents filled in questionnaires about quality of life and possible behavioral or emotional problems. The results revealed that all patients exhibited motor problems (ZNA). Most participants had further restricted oculomotor movements (ICARS). Age at injury and the full scale IQ were significantly positively correlated (Pearson correlation 0.779; p = 0.023). Conversely, no overall neuropsychological profile could be identified except for marginally reduced reaction times and susceptibility to interference. In addition, borderline results in semantic and phonemic word fluency tasks were apparent. A dysexecutive syndrome was diagnosed in one patient. However, verbal performance and reading abilities were non-pathologic in all participants. The patients reported having a good quality of life without major physical restrictions. Emotional disturbances and the presence of a mild cerebellar cognitive affective syndrome (as frequently described in adult patients) could only be confirmed in adolescents with vermis lesions. Nevertheless, in laboratory conditions, neuropsychological impairments were present in all patients. Heterogeneity of age at injury and exact lesion site may have led to interpersonal differences in neuropsychological outcome.


Sediment sources of Yan'gou watershed in the Loess Hilly region China under a certain rainstorm event.
At present the large scale vegetation restoration and intensive oil exploiting had brought huge influence on local environment in Yan'an region. the sediment yield data form series experiment plots and hydrological monitoring station in the Yan'an watershed after one rainfall event on July 2, 2005, which included sediment from different land uses (crop-land plot, vegetation plots, hard road surface) and 3 types roads(mountain-road brunches, mountain-road, and mountain-transport way) has been analyzed. Results showed that the erosion intensity of the 3 type roads was respectively 500 t/km(2), 3163 t/km(2), and 13500 t/km(2). The sediment from cropland and grass, shrub land was within 6-184 t/km(2). It stated that sediment from road area which only covered 1% of total area accounted for 42.3% of the total sediment yield, far beyond that from other uses of land. Sediment from grass-land and shrub-land, which covered 70.5% of watershed area, shared 26.7% of the total sediment. The further analysis showed that the 41.2% of total sediment could be detained by re-vegetation. On the contrary, that road constructing brought heavy sediment which offset the benefit of vegetation restore by 58.4%. the suggestion were to adjust our strategy from slope management to the road erosion mitigation Many studies have confirmed it is an important measure to return the steep slope farming land to green, and to restore vegetation in line with local conditions to prevent soil erosion in the Loess Plateau [1, 2]. To implement the measure in western region, the researches on "Grain for Green", returning farmland to forest and grassland, has become popular [3, 4]. Many scholars studied the effects of farming land and trees and grass land on soil water storage [2, 5]. Tang Keli stated that the slope land for farming was the main source of the sediment in Yellow river, and the maximum gradient slope for farming land use was 25° [6]. Many authorities not only pointed out that a far-reaching influence of land-use changes on the distribution of sediment source area but also put forward some new ideas about returning farming to green in Loess Plateau [7, 8]. However, we were still not sure the contribution of returning farmland to forest and grassland on reducing sediment yield of the valley and known it was difficult to identify its contribution to the total sediment yield. Analysis on the contribution of the stream channel and slope sediment yield had some results already [9, 10]. It was still too early to make clear the relationship between the sediment sources changes of the valley and the management. At present, Ecological restoration in the Loess Plateau caused the sediment form the slope land declining [3]. Due to human economic activities, the mountain road developed rapidly, it is inevitable that road erosion has been intensified [11]. A. Rijsdijk and LA Bruijnzeel (1991), [12] based the valley Konto observation, pointed out that although the rural road in the area accounts for only 3% of the area, but the impact on the sediment of this area was tremendous. Nyssen J, Moneryersons J. et al (2002) [13] also think that road without protection is one of the main sources of sediment. Many kinds of protective measures have great importance to the road erosion control. So attentions were paid to the study on the protection all kind of roads. Then what will happen to the soil erosion of the watershed, driven by the vegetation restoration and new road construction? What will happen to the proportion of sediment quality from slope land, road area and gully? A correct understanding of the sediment sources pattern of the typical watershed is of great significance on assessment the roles of vegetation to slope management and the roles of prevention the linear path erosion.


Hollow Co2P nanoflowers assembled from nanorods for ultralong cycle-life supercapacitors.
Hollow Co2P nanoflowers (Co2P HNFs) were successfully prepared via a one-step, template-free method. Microstructure analysis reveals that Co2P HNFs are assembled from nanorods and possess abundant mesopores and an amorphous carbon shell. Density functional theory calculations and electrochemical measurements demonstrate the high electrical conductivity of Co2P. Benefiting from the unique nanostructures, when employed as an electrode material for supercapacitors, Co2P HNFs exhibit a high specific capacitance, an outstanding rate capability, and an ultralong cycling stability. Furthermore, the constructed Co2P HNF//AC ASC exhibits a high energy density of 30.5 W h kg-1 at a power density of 850 W kg-1, along with a superior cycling performance (108.0% specific capacitance retained after 10 000 cycles at 5 A g-1). These impressive results make Co2P HNFs a promising candidate for supercapacitor applications.


Predictors of motor developmental milestones during the first year of life.
Studies suggest that both pre- and postnatal factors are predictors of age of attaining milestones in infancy. However, no studies evaluate the comparative strength of these predictors and the amount of the variance in development they explain. This study aimed to conduct a systematic evaluation of a broad selection of possible predictors of age at milestone attainment and to identify factors that explain significant inter-individual variance. Mothers of 5765 children of the Copenhagen Perinatal Cohort (1959-61) recorded 12 developmental milestones prospectively during the child's first year of life. Information on possible predictors was collected during pregnancy and at follow-up and was categorized into the domains: Family background, Pregnancy and delivery, Postnatal influences, and Postnatal growth. The domain Pregnancy and delivery contributed most of the explained variance in Overall mean of milestones (14.4%), with especially gestational age (β = -0.15; p ≤ 0.001) and birth weight (β = -0.16; p ≤ 0.001) being important predictors. Several individual factors, especially gestational age, birth weight, breastfeeding, having lived in a full-time institution, and weight and head increase in the first year, were significantly associated with milestone attainment in the first year of life. Variables within the domain of Pregnancy and delivery explained the largest proportion of variance in milestone attainment compared to the other domains. What is known: • Younger age at attainment of motor developmental milestones positively predicts cognitive outcomes in adulthood. • Both pre- and postnatal factors have been associated with age of attaining milestones in infancy. What is new: • First study to provide a systematic evaluation of a broad selection of predictors of infant milestone attainment. • Variables within the domain of Pregnancy and delivery, especially gestational age and birth weight, explained the largest proportion of variance in milestone attainment. • The variance explained by the predictors decreased time-dependently with later milestones.


The mean levels of adherence and factors contributing to non-adherence in patients on highly active antiretroviral therapy.
To determine the mean level of adherence and factors contributing to non-adherence in patients on Highly Active Antiretroviral Therapy (HAART). An observational study was done on 101 HIV/AIDS patients attending the Centre for HIV/ AIDS Research, Education and Services (CHARES) - University Hospital of the West Indies, between May 2006 and August 2006. A questionnaire was administered asking questions re: prescribed and actual dosing frequency and number of antiretroviral tablets for the previous week, reasons for nonadherence, duration of Highly Active Antiretroviral Therapy, age, employment status and level of education. Mean levels of adherence were calculated using self and social worker/nurse reported dosing frequency and number of tablets. Good adherence was defined as 95% or greater. Multiple regression analysis was used to determine factors impacting on adherence. Ninety-six patients were included for final analysis. Mean levels of adherence were as follows: 87.66%--self-report for tablets; 88.70%--self-report for dosing frequency; 87.02%--social worker/ nurse report for tablets; 88.10%--social worker/nurse report for dosing frequency. There were significant positive correlations between self and social worker/nurse reports using dosing frequency (Spearman Rho correlation coefficient 0.943, p = 0.01) or number of tablets (Spearman Rho correlation coefficient 0.955, p = 0.01). Adherence to self-reported number of tablets and dosing frequency were 58.4% and 56.4% respectively. Duration of HAART was found to have a significant negative correlation with the level of self-reported adherence to tablets (p = 0.002). Adherence to HAART is sub-optimum in patients at the CHARES. This must be urgently addressed to prevent the development of resistant HIV strains and treatment failure.


Tomographic diffractive microscopy and multiview profilometry with flexible aberration correction.
We have developed a tomographic diffractive microscope in reflection, which permits observation of sample surfaces with an improved lateral resolution, compared to a conventional holographic microscope. From the same set of data, high-precision measurements can be performed on the shape of the reflective surface by reconstructing the phase of the diffracted field. Doing so allows for several advantages compared to classical holographic interferometric measurements: improvement in lateral resolution, easier phase unwrapping, reduction of the coherent noise, combined with the high-longitudinal precision provided by interferometric phase measurements. We demonstrate these capabilities by imaging various test samples.


Work-related COVID-19 transmission in six Asian countries/areas: A follow-up study.
There is limited evidence of work-related transmission in the emerging coronaviral pandemic. We aimed to identify high-risk occupations for early coronavirus disease 2019 (COVID-19) local transmission. In this observational study, we extracted confirmed COVID-19 cases from governmental investigation reports in Hong Kong, Japan, Singapore, Taiwan, Thailand, and Vietnam. We followed each country/area for 40 days after its first locally transmitted case, and excluded all imported cases. We defined a possible work-related case as a worker with evidence of close contact with another confirmed case due to work, or an unknown contact history but likely to be infected in the working environment (e.g. an airport taxi driver). We calculated the case number for each occupation, and illustrated the temporal distribution of all possible work-related cases and healthcare worker (HCW) cases. The temporal distribution was further defined as early outbreak (the earliest 10 days of the following period) and late outbreak (11th to 40th days of the following period). We identified 103 possible work-related cases (14.9%) among a total of 690 local transmissions. The five occupation groups with the most cases were healthcare workers (HCWs) (22%), drivers and transport workers (18%), services and sales workers (18%), cleaning and domestic workers (9%) and public safety workers (7%). Possible work-related transmission played a substantial role in early outbreak (47.7% of early cases). Occupations at risk varied from early outbreak (predominantly services and sales workers, drivers, construction laborers, and religious professionals) to late outbreak (predominantly HCWs, drivers, cleaning and domestic workers, police officers, and religious professionals). Work-related transmission is considerable in early COVID-19 outbreaks, and the elevated risk of infection was not limited to HCW. Implementing preventive/surveillance strategies for high-risk working populations is warranted.


Hypertension in women: what is really known? The Women's Caucus, Working Group on Women's Health of the Society of General Internal Medicine.
To determine whether there is sufficient information in the medical literature to guide appropriate treatment of hypertensive women. Epidemiologic surveys of hypertension, clinical trials of antihypertensive therapy, and studies of selected adverse effects of antihypertensive agents were identified through a computerized search using MEDLINE and by identifying all studies cited in current medical textbooks as supporting evidence for the guidelines for the treatment of hypertensive individuals. All epidemiologic studies selected were cross-sectional or longitudinal, multicenter, population-based surveys. All clinical trials were large, randomized studies comparing one or more antihypertensive agents with a placebo or nonplacebo control group. Epidemiologic studies and clinical trials were reviewed to assess the quantity and quality of information available regarding important aspects of hypertension in women. Data pertaining to epidemiology, natural history, results of treatment, and two significant side effects of antihypertensive treatment were examined. The prevalence of hypertension is greater in black women than in black men and is about equal in white women and men. Because women outnumber men in the population, there are more hypertensive women than men. The attributable risk percent (the proportion of end points that could be eliminated by removing hypertension) for cardiovascular complications of hypertension is higher for women than men. Clinical trials show clear benefit of therapy for black women but no clear benefit for white women; some studies suggest that treatment of white women is harmful. Lipid profiles and their relation to ischemic heart disease differ for women and men; there is currently no information on the effects of antihypertensive agents on serum lipids in women. Few data have been published on the frequency of sexual dysfunction in treated hypertensive women. Hypertension in women and its related cardiovascular outcomes are a major public health problem. Clinical trials of antihypertensive therapy do not fully support current guidelines for the treatment of hypertensive women. Research concerning adverse effects of antihypertensive agents has largely excluded women from consideration; further studies are required to guide appropriate treatment.


Localized TWIST1 and TWIST2 basic domain substitutions cause four distinct human diseases that can be modeled in Caenorhabditis elegans.
Twist transcription factors, members of the basic helix-loop-helix family, play crucial roles in mesoderm development in all animals. Humans have two paralogous genes, TWIST1 and TWIST2, and mutations in each gene have been identified in specific craniofacial disorders. Here, we describe a new clinical entity, Sweeney-Cox syndrome, associated with distinct de novo amino acid substitutions (p.Glu117Val and p.Glu117Gly) at a highly conserved glutamic acid residue located in the basic DNA binding domain of TWIST1, in two subjects with frontonasal dysplasia and additional malformations. Although about one hundred different TWIST1 mutations have been reported in patients with the dominant haploinsufficiency Saethre-Chotzen syndrome (typically associated with craniosynostosis), substitutions uniquely affecting the Glu117 codon were not observed previously. Recently, subjects with Barber-Say and Ablepharon-Macrostomia syndromes were found to harbor heterozygous missense substitutions in the paralogous glutamic acid residue in TWIST2 (p.Glu75Ala, p.Glu75Gln and p.Glu75Lys). To study systematically the effects of these substitutions in individual cells of the developing mesoderm, we engineered all five disease-associated alleles into the equivalent Glu29 residue encoded by hlh-8, the single Twist homolog present in Caenorhabditis elegans. This allelic series revealed that different substitutions exhibit graded severity, in terms of both gene expression and cellular phenotype, which we incorporate into a model explaining the various human disease phenotypes. The genetic analysis favors a predominantly dominant-negative mechanism for the action of amino acid substitutions at this highly conserved glutamic acid residue and illustrates the value of systematic mutagenesis of C. elegans for focused investigation of human disease processes.


Use of peripheral quantitative computed tomography for densitometry of the femoral neck and spine in cynomolgus monkeys (Macaca fascicularis).
Quantitative computed tomography (QCT) allows for the separate densitometric examination of cortical and cancellous bone in vivo. With the new peripheral QCT (pQCT) instrument (the Norland/Stratec XCT-3000A), we evaluated the clinically relevant axial sites of spine and femoral neck in nonhuman primates in vivo. The reproducibility was good (coefficient of variation [CV] <3% at both sites for cortical, trabecular, and total bone mineral density [BMD]; CV 3%-7% for bone mineral content [BMC] and cross-sectional bone area). One hundred sixty intact female cynomolgus monkeys (M. fascicularis) were scanned at the femoral neck. There was less variability among monkeys in cortical BMD (mean 802 mg/mL, CV 6%) as opposed to trabecular BMD (mean 334 mg/mL, CV 28%) or transition zone BMD (mean 457 mg/mL, CV 12%). Scans were performed on lumbar vertebrae (L-4, L-5, and L-6) from five monkeys in vivo and ex vivo. Removal of soft tissue increased measured BMD. Decreasing voxel size from 0.4 mm to 0.2 mm increased measured BMD by diminishing the partial volume effect. Factor analysis demonstrated the expected relationships between pQCT parameters and physical measurement of bone mass and volume ex vivo. Preliminary results in eight ovariectomized and eight reproductively intact monkeys revealed a lower transition zone BMD at the femoral neck, and lower total BMD of the vertebral body in estrogen-deficient animals.


Chest X-ray screening examinations among French uranium miners: exposure estimation and impact on radon-associated lung cancer risk.
Medical surveillance of uranium miners can include periodic chest X-ray examinations. This study aimed to assess the X-ray exposure due to occupational health monitoring in the French cohort of uranium miners, and to test whether consideration of this additional radiation exposure impacts the excess risk of lung cancer death associated with radon exposure. X-ray exposure due to occupational health monitoring was estimated retrospectively based on review of a sample of miners' medical records and bibliographic data. Four exposure scenarios were designed, differing in their assumptions about the type of procedures performed, their frequency, and the lung dose delivered. Radon exposure and lung doses from exposure to α-particle emitters and external γ rays have previously been individually assessed. Exposure-risk and dose-risk relations were estimated by Poisson regression with a linear excess relative risk (ERR) model. The cohort included 5086 miners with a mean follow-up duration of 30.1 years. The mean number of chest X-ray examinations ranged from 15.1 in the lowest to 34 in the highest-exposure scenario, and produced a mean cumulative lung dose ranging from 4.6 to 34.2 mGy. The role of occupation-related imaging screening X-ray procedures in total equivalent lung dose appeared insignificant compared to α-emitter exposure. X-ray exposure was not associated with lung cancer mortality risk. The ERR associated with radon remained significantly positive when X-ray exposure was included in the multivariate analysis. X-ray exposure did not confound the exposure-risk relation between radon and lung cancer.


Position Paper on Asbestos of the Italian Society of Occupational Medicine.
The Position Paper (PP) on asbestos of the Italian Society of Occupational Medicine (SIML) aims at providing a tool to the occupational physician to address current diagnostic criteria and results of epidemiological studies, and their consequences in terms of preventive and evaluation actions for insurance, compensation and litigation. The PP was based on an extensive review of the scientific literature and was compiled by a Working Group comprising researchers who have contributed to the international literature on asbestos-related diseases, as well as occupational physicians with extensive experience in the evaluation of risks and the medical surveillance of workers currently and formerly exposed to asbestos. The PP was drafted and reviewed between 2017 and 2018; its final version was prepared according to the guidelines of AGREE Reporting Checklist. All the members of the Working Group subscribed to the document, which was eventually approved by SIML's Executive Committee. The first section addresses industrial hygiene issues, such as methods for environmental monitoring, advantages and limitations of different microscopy techniques, the potential role of microfibers and approaches for retrospective assessment of exposure, in particular in epidemiological studies. The second section reviews the biological effects of asbestos with particular attention to the diagnostic aspects of asbestosis, pleural changes, mesothelioma and lung cancer. In the following section the criteria of causal attribution are discussed, together with different hypotheses on the form of the risk functions, with a comparison of the opinions prevalent in the literature. In particular, the models of the risk function for mesothelioma were examined, in the light of the hypothesis of an acceleration or anticipation of the events in relation to the dose. The last section discusses topics of immediate relevance for the occupational physician, such as health surveillance of former exposed and of workers currently exposed in remediation activities.


Changes in biochemical characteristics and activities of ripening associated enzymes in mango fruit during the storage at different temperatures.
As a part of the study to explore the possible strategy for enhancing the shelf life of mango fruits, we investigated the changes in biochemical parameters and activities of ripening associated enzymes of Ashwina hybrid mangoes at 4-day regular intervals during storage at -10°C, 4°C, and 30 ± 1°C. Titratable acidity, vitamin C, starch content, and reducing sugar were higher at unripe state and gradually decreased with the increasing of storage time at all storage temperatures while phenol content, total soluble solid, total sugar, and nonreducing sugar contents gradually increased. The activities of amylase, α-mannosidase, α-glucosidase, and invertase increased sharply within first few days and decreased significantly in the later stage of ripening at 30 ± 1°C. Meanwhile polyphenol oxidase, β-galactosidase, and β-hexosaminidase predominantly increased significantly with the increasing days of storage till later stage of ripening. At -10°C and 4°C, the enzymes as well as carbohydrate contents of storage mango changed slightly up to 4 days and thereafter the enzyme became fully dormant. The results indicated that increase in storage temperature and time correlated with changes in biochemical parameters and activities of glycosidases suggested the suppression of β-galactosidase and β-hexosaminidase might enhance the shelf life of mango fruits.


Ruminal and intestinal degradability of distillers grains plus solubles varies by source.
Currently in the dairy industry, there is a concern about the variability in the nutrient content among sources of distillers grains plus solubles (DG), but little research has evaluated the variability in metabolizable AA among sources. The ruminal degradability of crude protein (CP) in soybean meal (SBM), dried DG from 5 sources (DG1, DG2, DG3, DG4, and DG5), and 1 source of wet DG (WDG) were determined using 2 lactating ruminally cannulated Holstein cows. Feeds were incubated in the rumen for 3, 6, 12, 18, 24, and 36 h. Intestinal CP digestibility via pepsin and pancreatin and AA profiles were measured on residue from 12-h ruminal incubation of feeds. Ruminal undegradable protein (RUP) was less for SBM (46.4%) than for DG. The WDG (53.6%) had less RUP than dried DG. The RUP concentrations of DG3 (59.1%) and DG5 (60.3%) were lower than DG1 (71.7%) and DG4 (67.5%), with DG1 having more than DG2 (63.7%) and DG4. Intestinal digestibility of RUP was greater for SBM (86.7%) than DG. The DG2 (76.8%) and DG3 (74.2%) had greater intestinal digestibility compared with DG1 (59.2%), DG4 (63.0%), and DG5 (68.1%). The intestinal digestibility in WDG (65.8%) was similar to all other DG except for DG1, which was lower. Total digestibility of CP was greater in SBM (93.9%) compared with DG. Among the DG sources, the CP in DG2 (85.3%) and DG3 (84.9%) was more digestible compared with DG1 (70.7%), DG4 (74.9%), and DG5 (80.8%) but not WDG (81.9%). Based on the milk protein score (MPS), which is an estimate of the proportion of milk protein that a protein source can sustain until the first limiting AA is depleted, Met was the first limiting AA in SBM and Lys in DG. The concentrations of essential AA in the RUP were not different among DG sources, but the greater MPS in WDG (0.306) compared with the dried DG (0.240) sources indicated that WDG may have been the more ideal RUP source; but, the MPS of the metabolizable protein indicated that the protein quality of WDG was similar to that in DG2, DG3, and DG5. In conclusion, protein degradability and digestibility differed greatly among DG sources, but these differences were actually not as prominent in the concentrations of metabolizable AA and MPS among these sources.


Language mapping in less than 15 minutes: real-time functional MRI during routine clinical investigation.
Neurosurgical interventions often require the presurgical determination of language dominance or mapping of language areas. Results obtained by fMRI are closely correlated with invasive procedures such as electrical stimulation mapping or the intracarotid amobarbital test. However, language fMRI is not used routinely, because postprocessing is time-consuming. We utilized a real-time analysis software installed directly on the MR console computer and SPM99 as reference postprocessing software. We assessed the reliability of the immediate determination of language dominance based on individual activation maps by comparing the results of the visual analysis of images derived from conventional postprocessing with those produced by the real-time tool. All images were rated independently by six senior neurologists blinded to other data. We validated the robustness of the real-time method statistically by comparing global and regional lateralization indices derived from real-time and postprocessing analysis. Functional MRI was performed with a standard 1.5-T whole-body scanner. Brain activity was contrasted between an alternating semantic judgment and letter matching task. Twelve right-handed, healthy control subjects and 12 consecutive patients with drug-resistant, localization-related epilepsy were investigated. The semantic condition induced almost invariably left hemispheric activations in Broca's area, the premotor cortex, the dorsolateral prefrontal cortex, and the temporoparietal region. Although real-time analysis reduced noise less effectively than SPM99, visual ratings and lateralization indices produced highly concordant results with both methods. In conclusion, real-time fMRI, as used here, allowed reliable language lateralization and mapping in less than 15 min during routine clinical MRI investigation with no need for postprocessing.


Serum neurofilament levels correlate with retinal nerve fiber layer thinning in multiple sclerosis.
Serum neurofilament light chain (sNfL) levels and peripapillary retinal nerve fiber layer (pRNFL) are both emerging biomarkers of neuro-axonal damage in multiple sclerosis (MS). However, data on the relation between sNfL and pRNFL are scarce. We aimed to determine the relation of sNfL levels with pRNFL thinning in a large cohort of relapsing-remitting (RR) MS patients. We identified 80 patients from a prospective, 3-year observational study on retinal changes in RRMS with annual blood samples available. sNfL levels were measured using single-molecule array (SimoaTM) assay. Annualized loss of pRNFL (aLpRNFL) was determined by individual linear regression models. Correlations between single and repeated sNfL levels and aLpRNFL were analyzed using multivariate linear regression and mixed-effect models. After correction for sex, age, and baseline sNfL, an sNfL increase of 10 pg/mL was associated with an aLpRNFL of -0.7 µm (95% confidence interval (CI): (-1.3, -0.2), p < 0.001). Patients with ⩾2 sNfL measurements >75th percentile displayed higher aLpRNFL (2.2 µm, standard deviation (SD) 0.6) compared to patients with no sNfL measure >75th percentile (0.4 µm, SD 0.2, p < 0.001). Between 15% and 20% of the aLpRNFL variance could be predicted from sNfL levels. sNfL levels contribute to the prediction of retinal thinning in patients with RRMS, strengthening its value as a biomarker of neuro-axonal damage.


Opening Pandora's box?: ethical issues in prenatal whole genome and exome sequencing.
The development of genomic approaches to prenatal testing such as whole genome and exome sequencing offers the potential for a better understanding of prenatal structural anomalies in the fetus and ultimately for improved patient care and more informed reproductive decision making. In addition to the scientific and clinical challenges of achieving this, the introduction of new reproductive technologies also presents a number of ethical problems. The successful and appropriate development and introduction of prenatal genomics into clinical practice require these problems to be identified, understood and carefully analysed in the development of models of good ethical practice. We conducted a critical review of the existing literature on ethical issues in prenatal genomics. We identified and discussed five areas of particular concern: valid consent, management and feed-back of information, responsibilities of health professionals, priority setting and resources and duties towards the future child. There is a need for further discussion of the issues we have outlined here, and we hope that this brief summary of ethical arguments in the literature encourages researchers, clinicians, patients and scientists to engage in further discussion of these and other important issues raised by prenatal genomics. © 2017 The Authors. Prenatal Diagnosis published by John Wiley & Sons, Ltd.


Immunotherapy in Bladder Cancer.
With the advent of Bacille Calmette Guerin (BCG), bladder cancer was one of the earliest cancers where the concept of immunotherapy was utilized. While this is true, recent advances in the use of immunotherapy are enabling oncologists to expand the armamentarium for the treatment of bladder cancer. Unacceptable side effects and failure to produce a durable response with the use of chemotherapeutic agents in bladder cancer has led to the evaluation of more targeted and personalized approaches. Increased understanding of the underlying carcinogenesis of bladder cancer, coupled with the ability to engineer targeted agents implicated in bladder cancer associated pathways has provided new avenues for the management of this disease. Newer immunotherapeutic approaches have generated a great deal of interest in bladder cancer along with other diseases. In this article we will focus on various forms of immunotherapies that may have a therapeutic potential in bladder cancer. We will briefly review the current status of "non-targeted" immunotherapeutic agents like BCG, interferons and interleukins in bladder cancer. But the main focus of this article is to discuss the emerging role of "targeted" immunotherapeutic agents like cytotoxic T cell lymphocyte associated protein-4 blocking antibody and programmed death pathway blocking antibodies in localized or metastatic bladder cancer.


Kainic acid-induced neuronal death is associated with DNA damage and a unique immediate-early gene response in c-fos-lacZ transgenic rats.
Previously, we established that persistent upregulation of c-fos expression preceded kainic acid (KA)-induced neuronal death in mice. To discriminate between events that are products of the seizures elicited by KA and those that are specifically associated with its neurotoxic actions, we have examined the expression of cellular immediate-early genes (cIEGs) following KA or pentylenetetrazol (PTZ) treatment in c-fos-lacZ transgenic rats. While both chemoconvulsants elicit seizures, only KA causes selective neuronal death. Following treatment of transgenic rats with KA there was a protracted expression of Fos-lacZ that lasted for 2-3 d. In contrast, PTZ elicited a transient increase in the transgene product that lasted about 6 hr. Normally, Fos and Fos-lacZ were detected only in neuronal nuclei. However, 6 hr following kainic acid (but not PTZ) administration, beta-galactosidase activity appeared in the cytoplasm of neurons within vulnerable regions (as determined by the terminal transferase biotinylated-UTP nick end labeling (TUNEL) procedure). Like c-fos, transcripts for other cIEGs were elevated for longer periods in the KA-treated rat hippocampus. In addition, fra-1 and fra-2 were only induced in the KA-treated rat. These changes in mRNA levels were paralleled by a sustained increase in AP-1 DNA binding activity. Thus, quantitative and qualitative changes in AP-1 DNA binding complexes accompany neurotoxic cell death that are not observed following seizures.


Deletion of Apoptosis Inhibitor F1L in Vaccinia Virus Increases Safety and Oncolysis for Cancer Therapy.
Vaccinia virus (VACV) possesses a great safety record as a smallpox vaccine and has been intensively used as an oncolytic virus against various types of cancer over the past decade. Different strategies were developed to make VACV safe and selective to cancer cells. Leading clinical candidates, such as Pexa-Vec, are attenuated through deletion of the viral thymidine kinase (TK) gene, which limits virus growth to replicate in cancer tissue. However, tumors are not the only tissues whose metabolic activity can overcome the lack of viral TK. In this study, we sought to further increase the tumor-specific replication and oncolytic potential of Copenhagen strain VACV ΔTK. We show that deletion of the anti-apoptosis viral gene F1L not only increases the safety of the Copenhagen ΔTK virus but also improves its oncolytic activity in an aggressive glioblastoma model. The additional loss of F1L does not affect VACV replication capacity, yet its ability to induce cancer cell death is significantly increased. Our results also indicate that cell death induced by the Copenhagen ΔTK/F1L mutant releases more immunogenic signals, as indicated by increased levels of IL-1β production. A cytotoxicity screen in an NCI-60 panel shows that the ΔTK/F1L virus induces faster tumor cell death in different cancer types. Most importantly, we show that, compared to the TK-deleted virus, the ΔTK/F1L virus is attenuated in human normal cells and causes fewer pox lesions in murine models. Collectively, our findings describe a new oncolytic vaccinia deletion strain that improves safety and increases tumor cell killing.


Genetic evidence of heterogeneity in intrahepatic cholestasis of pregnancy.
The aim of this study was to investigate the genetic aetiology of intrahepatic cholestasis of pregnancy (ICP) and the impact of known cholestasis genes (BSEP, FIC1, and MDR3) on the development of this disease. Sixty nine Finnish ICP patients were prospectively interviewed for a family history of ICP, and clinical features were compared in patients with familial ICP (patients with a positive family history, n=11) and sporadic patients (patients with no known family history of ICP, n=58). For molecular genetic analysis, 16 individuals from two independently ascertained Finnish ICP families were genotyped for the flanking markers for BSEP, FIC1, and MDR3. The pedigree structures in 16% (11/69) of patients suggested dominant inheritance. Patients with familial ICP had higher serum aminotransferase levels and a higher recurrence risk (92% v 40%). Both segregation of haplotypes and multipoint linkage analysis excluded BSEP, FIC1, and MDR3 genes in the studied pedigrees. Additionally, the MDR3 gene, previously shown to harbour mutations in ICP patients, was negative for mutations when sequenced in four affected individuals from the two families. These results support the hypothesis that the aetiology of ICP is heterogeneous and that ICP is due to a genetic predisposition in a proportion of patients. The results of molecular genetic analysis further suggest that the previously identified three cholestasis genes are not likely to be implicated in these Finnish ICP families with dominant inheritance.


Association of Ultrafiltration Rate with Mortality in Incident Hemodialysis Patients.
Ultrafiltration rate (UFR) appears to be associated with mortality in prevalent hemodialysis (HD) patients. However, the association of UFR with mortality in incident HD patients remains unknown. We examined a US cohort of 110,880 patients who initiated HD from 2007 to 2011. Baseline UFR was divided into 5 groups (<4, 4 to <6, 6 to <8, 8 to <10, and ≥10 mL/h/kg body weight [BW]). We examined predictors of higher baseline UFR using logistic regression and the association of baseline UFR and all-cause and cardiovascular (CV) mortality using Cox proportional hazard models with adjustments for demographics, comorbidities, and markers of malnutrition-inflammation-cachexia syndrome. Patients were 63 ± 15 years, with 43% women, 32% African Americans, and had a mean baseline UFR of 7.5 ± 3.1 mL/h/kg BW. In the fully adjusted logistic regression models, factors associated with higher UFR (≥7.5 mL/h/kg BW) included Hispanic ethnicity, diabetes, and higher dietary protein intake. There was a linear association between UFR and all-cause and CV mortality, where UFR ≥10 mL/h/kg BW (reference UFR 6-<8 mL/h/kg BW) conferred the highest risk in both unadjusted (HR 1.15 [95% CI 1.10-1.19]) and adjusted models (HR 1.23 [95% CI 1.16-1.31]). The linear association with all-cause mortality remained consistent across strata of age, urine volume, and treatment time. Higher UFR is independently associated with higher all-cause and CV mortality in incident HD patients. Clinical trials are warranted to examine the effects of lowering UFR on outcomes.


Association Between Patient Characteristics and Treatment Procedure Among Patients With Uterine Leiomyomas.
To analyze the association between patient characteristics and the probability of undergoing any uterine-sparing procedure (endometrial ablation, myomectomy, and uterine artery embolization) compared with hysterectomy as the first uterine leiomyoma (index) procedure and the probability of undergoing a specific uterine-sparing procedure. We conducted a retrospective analysis using a commercial insurance claims database containing more than 13 million enrollees annually. Based on the index procedure performed 2004-2009, women were classified into one of the four procedure cohorts. Eligible women were aged 25-54 years on the index date, continuously insured through 1-year baseline and 1-year follow-up, and had a baseline uterine leiomyoma diagnosis. Logistic regression was used to assess the association between patient characteristics and leiomyoma procedure. The study sample comprised 96,852 patients (endometrial ablation=12,169; myomectomy=7,039; uterine artery embolization=3,835; and hysterectomy=73,809). Patient characteristics associated with undergoing any uterine-sparing procedure compared with hysterectomy included health maintenance organization health plan enrollment, Northeast region residence, the highest income and education quintiles based on zip code, an age-race interaction, and baseline diagnoses including menstrual disorders, pelvic pain, anemia, endometriosis, genital prolapse, and infertility. Among those who had a uterine-sparing procedure, characteristics associated with undergoing uterine artery embolization or endometrial ablation compared with myomectomy included increasing age, being from the Midwest relative to the Northeast, and certain baseline conditions including menstrual disorder, pelvic pain, endometriosis, and infertility. Both clinical and nonclinical factors were associated with the receipt of alternatives to hysterectomy for uterine leiomyomas in commercially insured women.


Using Shapes of COVID-19 Positive Patient-Specific Trajectories for Mortality Prediction.
Machine learning can be used to identify relevant trajectory shape features for improved predictive risk modeling, which can help inform decisions for individualized patient management in intensive care during COVID-19 outbreaks. We present explainable random forests to dynamically predict next day mortality risk in COVID -19 positive and negative patients admitted to the Mount Sinai Health System between March 1st and June 8th, 2020 using patient time-series data of vitals, blood and other laboratory measurements from the previous 7 days. Three different models were assessed by using time series with: 1) most recent patient measurements, 2) summary statistics of trajectories (min/max/median/first/last/count), and 3) coefficients of fitted cubic splines to trajectories. AUROC and AUPRC with cross-validation were used to compare models. We found that the second and third models performed statistically significantly better than the first model. Model interpretations are provided at patient-specific level to inform resource allocation and patient care.


The crystal structure of the N-terminal region of BUB1 provides insight into the mechanism of BUB1 recruitment to kinetochores.
The interaction of the central mitotic checkpoint component BUB1 with the mitotic kinetochore protein Blinkin is required for the kinetochore localization and function of BUB1 in the mitotic spindle assembly checkpoint, the regulatory mechanism of the cell cycle that ensures the even distribution of chromosomes during the transition from metaphase to anaphase. Here, we report the 1.74 angstroms resolution crystal structure of the N-terminal region of BUB1. The structure is organized as a tandem arrangement of three divergent units of the tetratricopeptide motif. Functional assays in vivo of native and site-specific mutants identify the residues of human BUB1 important for the interaction with Blinkin and define one region of potential therapeutic interest. The structure provides insight into the molecular basis of Blinkin-specific recognition by BUB1 and, on a broader perspective, of the mechanism that mediates kinetochore localization of BUB1 in checkpoint-activated cells.


Doxycycline prophylaxis of travelers' diarrhea in Honduras, an area where resistance to doxycycline is common among enterotoxigenic Escherichia coli.
Daily doxycycline (DX), known to be effective prophylaxis against travelers' diarrhea (TD) in areas of the world where enterotoxigenic Escherichia coli (ETEC) are sensitive to the drug, has not been extensively studied in geographic areas where antibiotic resistance is common. Therefore we studied 44 U.S. Peace Corps Volunteers during their first 5 weeks in Honduras, which is such an area. During the first 3 weeks, volunteers took daily either 100 mg DX or placebo (PL) in a double-blind, randomized fashion. All 22 taking PL developed TD during the first 3 weeks, compared to 7 of 22 (32%) taking DX (P less than 0.001; 68% protection). ETEC were isolated from 39% of episodes of TD. From the PL group, ETEC from 7 of 13 stool samples (54%) were resistant to DX, whereas from the DX group, ETEC from 10 of 11 stool samples were resistant (P less than 0.05). TD that developed in persons taking DX was also found to be less severe, as judged by length of illness (P less than 0.01) and frequency of stools (P less than 0.05). This study demonstrates that DX 1) significantly prevents TD even in areas where antibiotic resistance is common, although it does not prevent TD caused by docycycline -resistant ETEC, and 2) significantly diminishes the severity of illness.


Th2-like CD8+ T cells showing B cell helper function and reduced cytolytic activity in human immunodeficiency virus type 1 infection.
We analyzed at clonal level the functional profile of circulating or skin-infiltrating T lymphocytes from two individuals infected with the human immunodeficiency virus type 1 (HIV-1), suffering from a Job's-like syndrome (eczematous dermatitis, recurrent skin and sinopulmonary infections, and hypergammaglobulinemia E) and showing virtually no circulating CD4+ T cells. Most of the CD3+ T cell clones generated from both patients were CD4- CD8+ TCR alpha beta +. The others were CD4- CD8- TCR alpha beta + which exhibited reduced mRNA expression for the CD8 molecule or no mRNA expression for either CD4 or CD8 molecules. The great majority of both CD4- CD8+ and CD4- CD8- did not produce interferon (IFN) gamma and exhibited reduced cytolytic activity. Rather, most of them produced large amounts of both interleukin (IL) 4 and IL-5 and provided B cell helper function for IgE synthesis. These data suggest that a switch of cytolytic CD8+ T cells showing a Th1-like cytokine secretion profile to cells that make Th2-type cytokines, exhibit reduced cytolytic potential, and provide B cell helper function can occur in the course of HIV-1 infection. These cells may contribute to the reduced defense against viral infections and intracellular parasites and account for the elevated IgE serum levels, eosinophilia, and the allergic-like clinical manifestations seen in a proportion of HIV-1-infected individuals.