An integrated multi-study analysis of intra-subject variability in cerebrospinal fluid amyloid-β concentrations collected by lumbar puncture and indwelling lumbar catheter.
Amyloid-β (Aβ) has been investigated as a diagnostic biomarker and therapeutic drug target. Recent studies found that cerebrospinal fluid (CSF) Aβ fluctuates over time, including as a diurnal pattern, and increases in absolute concentration with serial collection. It is currently unknown what effect differences in CSF collection methodology have on Aβ variability. In this study, we sought to determine the effect of different collection methodologies on the stability of CSF Aβ concentrations over time. Grouped analysis of CSF Aβ levels from multiple industry and academic groups collected by either lumbar puncture (n=83) or indwelling lumbar catheter (n=178). Participants were either placebo or untreated subjects from clinical drug trials or observational studies. Participants had CSF collected by lumbar puncture or lumbar catheter for quantitation of Aβ concentration by enzyme linked immunosorbent assay. Data from all sponsors was converted to percent of the mean for Aβ40 and Aβ42 for comparison. Repeated measures analysis of variance was performed to assess for factors affecting the linear rise of Aβ concentrations over time. Analysis of studies collecting CSF via lumbar catheter revealed tremendous inter-subject variability of Aβ40 and Aβ42 as well as an Aβ diurnal pattern in all of the sponsors' studies. In contrast, Aβ concentrations from CSF samples collected at two time points by lumbar puncture showed no significant differences. Repeated measures analysis of variance found that only time and draw frequency were significantly associated with the slope of linear rise in Aβ40 and Aβ42 concentrations during the first 6 hours of collection. Based on our findings, we recommend minimizing the frequency of CSF draws in studies measuring Aβ levels and keeping the frequency standardized between experimental groups. The Aβ diurnal pattern was noted in all sponsors' studies and was not an artifact of study design. Averaging Aβ concentrations at each time point is recommended to minimize the effect of individual variability. Indwelling lumbar catheters are an invaluable research tool for following changes in CSF Aβ over 24-48 hours, but factors affecting Aβ concentration such as linear rise and diurnal variation need to be accounted for in planning study designs.


Lifestyle-Related Factors and Total Mortality in a Mediterranean Prospective Cohort.
Lifestyle-related habits have a strong influence on morbidity and mortality worldwide. This study investigates the association between a multidimensional healthy lifestyle score and all-cause mortality risk, including in the score some less-studied lifestyle-related factors. Participants (n=20,094) of the Seguimiento Universidad de Navarra cohort were followed up from 1999 to 2018. The analysis was conducted in 2019. A 10-point healthy lifestyle score previously associated with a lower risk of major cardiovascular events was applied, assigning 1 point to each of the following items: never smoking, moderate-to-high physical activity, moderate-to-high Mediterranean diet adherence, healthy BMI, moderate alcohol consumption, avoidance of binge drinking, low TV exposure, short afternoon nap, time spent with friends, and working ≥40 hours per week. During a median follow-up of 10.8 years, 407 deaths were documented. In the multivariable adjusted analysis, the highest category of adherence to the score (7-10 points) showed a 60% lower risk of all-cause mortality than the lowest category (0-3 points) (hazard ratio=0.40, 95% CI=0.27, 0.60, p<0.001 for trend). In analyses of the healthy lifestyle score as a continuous variable, for each additional point in the score, a 18% relatively lower risk of all-cause mortality was observed (adjusted hazard ratio=0.82, 95% CI=0.76, 0.88). Adherence to a healthy lifestyle score, including some less-studied lifestyle-related factors, was longitudinally associated with a substantially lower mortality rate in a Mediterranean cohort. Comprehensive health promotion should be a public health priority.


Long-term clinical outcome and graft patency of radial artery and saphenous vein grafts in multiple arterial revascularization.
The long-term benefits of multiple arterial revascularization (MAR) in coronary artery bypass grafting remain uncertain. The aim of this study was to investigate the clinical outcome, graft patency, and need for subsequent target revascularization of radial artery (RA) versus saphenous vein graft in patients undergoing MAR in both patient- and graft-specific analyses. Between 2001 and 2016, we followed 1654 patients over a median of 7.4 years in a prospective, longitudinal study. Major adverse cardiac and cerebrovascular events, graft patency, and need for revascularization were assessed through clinical manifestation, coronary angiography, or coronary computed tomography and analyzed with propensity score-adjusted Cox regression, general estimating equation, and competing risk models. Bilateral internal thoracic artery (BITA) grafting was performed in 910 patients (55.0%), and 744 patients (45.0%) received a left internal thoracic artery graft together with at least 1 RA graft. Patients receiving BITA, of whom 187 received an additional RA, showed improved survival (hazard ratio, 0.57; 95% confidence interval [CI], 0.38-0.86; P = .009), major adverse cardiac and cerebrovascular event-free survival (hazard ratio, 0.33; 95% CI, 0.23-0.46; P < .001), and lower need for repeat revascularization (subhzhard ratio, 0.59; 95% CI, 0.39-0.90; P = .015). In a subgroup of 512 patients, comparing 419 RA with 487 saphenous vein grafts, RA grafting showed a lower risk for graft occlusion (odds ratio, 0.59; 95% CI, 0.47-0.73; P < .001) and target revascularization (subhazard ratio, 0.58; 95% CI, 0.43-0.78; P < .001). MAR with BITA and RA grafting revealed to be the recommended strategy in coronary artery bypass grafting to achieve long-term beneficial results. The use of saphenous vein graft showed less favorable outcomes regarding patency and the need for target-vessel revascularization.


Evaluation of psychometric properties and factorial structure of the pre-school child behaviour checklist at the Kenyan Coast.
Behavioural/emotional problems may be common in preschool children living in resource-poor settings, but assessment of these problems in preschool children from poor areas is challenging owing to lack of appropriate behavioural screening tools. The child behaviour checklist (CBCL) is widely known for its reliability in identifying behavioural/emotional problems in preschool children, but it has not been validated for use in sub-Saharan Africa. With permission from developers of CBCL, we translated this tool into Ki-Swahili and adapted the items to make them culturally appropriate and contextually relevant and examined the psychometric properties of the CBCL, particularly reliability, validity and factorial structure in a Kenyan community preschool sample of 301 children. It was also re-administered after 2 weeks to 38 randomly selected respondents, for the purpose of evaluating retest reliability. To evaluate inter-informant reliability, the CBCL was administered to 46 respondents (17 alternative caretakers and 29 fathers) alongside the child's mother. Generalised linear model was used to measure associations with behavioural/emotional scores. We used structural equation modelling to perform a confirmatory factor analysis to examine the seven-syndrome CBCL structure. During the first phase we found that most of the items could be adequately translated and easily understood by the participants. The inter-informant agreement for CBCL scores was excellent between the mothers and other caretakers [Pearson's correlation coefficient (r) = 0.89, p < 0.001] and fathers (r = 0.81; p < 0.001). The test-retest reliability was acceptable (r = 0.76; p < 0.001). The scale internal consistency coefficients were excellent for total problems [Cronbach's alpha (α) = 0.95] and between good and excellent for most CBCL sub-scales (α = 0.65-0.86). Behavioural/emotional scores were associated with pregnancy complications [adjusted beta coefficient (β) = 0.44 (95 % CI, 0.07-0.81)] and adverse perinatal events [β = 0.61 (95 % CI, 0.09-1.13)] suggesting discriminant validity of the CBCL. Most fit indices for the seven-syndrome CBCL structure were within acceptable range, being <0.09 for root mean squared error of approximation and >0.90 for Tucker-Lewis Index and Comparative Fit Index. The CBCL has good psychometric properties and the seven-syndrome structure fits well with the Kenyan preschool children suggesting it can be used to assess behavioural/emotional problems in this rural area.


Small-sample blood culture method for identification of bacteria in central arterial and peripheral blood.
A blood culture technique that utilized small arterial blood samples or peripheral capillary blood was tested in beagle dogs and pig-tailed macaque monkeys. A bolus of 2.0 x 10(7)Escherichia coli (ATCC 25922) was injected intravenously into five animals of each species. Blood samples were taken before injection of the organisms and 10, 15, 20, 30, 60, and 120 min after injection. Arterial blood samples (2.0 and 0.2 ml) and peripheral capillary samples (0.14 ml) were taken at each sampling time. Pour plates were prepared from arterial blood for colony counts. All three blood sampling methods were equally effective in detecting sepsis when 10 or more organisms per ml of blood were present. Below this level, the 2.0-ml sample was more effective. Contamination of the peripheral sample with air or skin contaminants was a problem.


The Society for Immunotherapy of Cancer statement on best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) staining and validation.
The interaction between the immune system and tumor cells is an important feature for the prognosis and treatment of cancer. Multiplex immunohistochemistry (mIHC) and multiplex immunofluorescence (mIF) analyses are emerging technologies that can be used to help quantify immune cell subsets, their functional state, and their spatial arrangement within the tumor microenvironment. The Society for Immunotherapy of Cancer (SITC) convened a task force of pathologists and laboratory leaders from academic centers as well as experts from pharmaceutical and diagnostic companies to develop best practice guidelines for the optimization and validation of mIHC/mIF assays across platforms. Representative outputs and the advantages and disadvantages of mIHC/mIF approaches, such as multiplexed chromogenic IHC, multiplexed immunohistochemical consecutive staining on single slide, mIF (including multispectral approaches), tissue-based mass spectrometry, and digital spatial profiling are discussed. mIHC/mIF technologies are becoming standard tools for biomarker studies and are likely to enter routine clinical practice in the near future. Careful assay optimization and validation will help ensure outputs are robust and comparable across laboratories as well as potentially across mIHC/mIF platforms. Quantitative image analysis of mIHC/mIF output and data management considerations will be addressed in a complementary manuscript from this task force.


Expression of cornulin in tongue squamous cell carcinoma.
The aim of the study is to identify cornulin (CRNN) protein expression associated with advancement of tongue squamous cell carcinoma (TSCC). A comparison of addictive (containing potential carcinogens) versus non-addiction causative agents was expected to allow detection of differences in CRNN expression associated with TSCC. Bespoke tissue microarrays (TMAs) were prepared and immunohistochemistry (IHC) performed to determine the changes in CRNN expression in epithelial cells of node-negative (pN-), node-positive (pN+) TSCC and non-cancer patients' oral tissues. TMAs were validated by performing IHC on whole diagnostic tissues. Chi-square test or Fisher's-exact tests were used to establish significant expression differences. Analogous analyses were performed for biomarkers previously associated with TSCC, namely collagen I alpha 2 (COL1A2) and decorin (DCN) to compare the significance of CRNN. Keratinisation and its level (low, extensive) were studied in relation to CRNN so that the extent of squamous differentiation could better be assessed. IHC immunoreactive score (IRS) clustered the patients based on weak/moderate (Low (IRS ≤ +3)) or strong (High (IRS ≥ +4)) expression groups. A low expression was observed in a larger number of patients in control proteins COL1A2 (77.3%), DCN (87.5%) and target protein CRNN (52.3%), respectively. Low CRNN expression was observed in TSCC where nodes were involved (pN+: mean 1.4 ± 2.1) (p = 0.248). Keratinisation (%) was low (0% ≤ 50%) in 42.2% and extensive (1% ≥ 50.0%) in 57.8% patients. In conclusion, our study suggested that Low CRNN expression was associated with grade and lymph node metastasis in TSCC. CRNN expression is independent of addiction, however potentially carcinogenic addictive substances might be aiding in the disease progression.


Scanning the periphery.
Companies often face new rivals, technologies, regulations, and other environmental changes that seem to come out of left field. How can they see these changes sooner and capitalize on them? Such changes often begin as weak signals on what the authors call the periphery, or the blurry zone at the edge of an organization's vision. As with human peripheral vision, these signals are difficult to see and interpret but can be vital to success or survival. Unfortunately, most companies lack a systematic method for determining where on the periphery they should be looking, how to interpret the weak signals they see, and how to allocate limited scanning resources. This article provides such a method-a question-based framework for helping companies scan the periphery more efficiently and effectively. The framework divides questions into three categories: learning from the past (What have been our past blind spots? What instructive analogies do other industries offer? Who in the industry is skilled at picking up weak signals and acting on them?); evaluating the present (What important signals are we rationalizing away? What are our mavericks, outliers, complainers, and defectors telling us? What are our peripheral customers and competitors really thinking?); and envisioning the future (What future surprises could really hurt or help us? What emerging technologies could change the game? Is there an unthinkable scenario that might disrupt our business?). Answering these questions is a good first step toward anticipating problems or opportunities that may appear on the business horizon. The article concludes with a self-test that companies can use to assess their need and capability for peripheral vision.


Heat shock protein 70 (hsp70) as a major target of the antibody response in patients with pulmonary cryptococcosis.
Cryptococcus neoformans causes infection in individuals with defective T cell function, such as AIDS, as well as without underlying disease. It has been suggested that humoral as well as cellular immunity might play an important role in the immune response to C. neoformans infection. We have recently shown, using immunoblotting, that the 70-kD hsp family of C. neoformans was the major target molecule of the humoral response in murine pulmonary cryptococcosis. In this study we also used immunoblotting to define the antibody responses in the sera of 24 patients with pulmonary cryptococcosis: 21 proven and three suspected diagnoses. Anti-C. neoformans hsp70 antibody was detected in 16 of 24 (66.7%) patients with pulmonary cryptococcosis. Fourteen of 17 (82.3%) patients with high antigen titres (> or = 1:8) and two of seven (28.6%) patients with low titres (< or = 1:4) had detectable levels of anti-hsp70 antibody. Sera from patients positive for anti-hsp70 antibody showed high titres in the Eiken latex agglutination test for the detection of serum cryptococcal antigen. Our results indicate that the 70-kD hsp family from C. neoformans appears to be a major target molecule of the humoral response, not only in murine pulmonary cryptococcosis, but also in human patients with pulmonary cryptococcosis.


Recreational Physical Activity and Risk of Incident Knee Osteoarthritis: An International Meta-Analysis of Individual Participant-Level Data.
The effect of physical activity on the risk of developing knee osteoarthritis (OA) is unclear. We undertook this study to examine the relationship between recreational physical activity and incident knee OA outcomes using comparable physical activity and OA definitions. Data were acquired from 6 global, community-based cohorts of participants with and those without knee OA. Eligible participants had no evidence of knee OA or rheumatoid arthritis at baseline. Participants were followed up for 5-12 years for incident outcomes including the following: 1) radiographic knee OA (Kellgren-Lawrence [K/L] grade ≥2), 2) painful radiographic knee OA (radiographic OA with knee pain), and 3) OA-related knee pain. Self-reported recreational physical activity included sports and walking/cycling activities and was quantified at baseline as metabolic equivalents of task (METs) in days per week. Risk ratios (RRs) were calculated and pooled using individual participant data meta-analysis. Secondary analysis assessed the association between physical activity, defined as time (hours per week) spent in recreational physical activity and incident knee OA outcomes. Based on a total of 5,065 participants, pooled RR estimates for the association of MET days per week with painful radiographic OA (RR 1.02 [95% confidence interval (95% CI) 0.93-1.12]), radiographic OA (RR 1.00 [95% CI 0.94-1.07]), and OA-related knee pain (RR 1.00 [95% CI 0.96-1.04]) were not significant. Similarly, the analysis of hours per week spent in physical activity also showed no significant associations with all outcomes. Our findings suggest that whole-body, physiologic energy expenditure during recreational activities and time spent in physical activity were not associated with incident knee OA outcomes.


Association between the altitude of residential areas and clinical diagnosis of chronic gastritis in ambulatory patients of Peru: A cross-sectional analytic study.
Chronic gastritis is one of the most common diseases in the population. Several factors influence its appearance; however, the effect of high altitude has not been studied thoroughly. To determine the association between the altitude of the residential area and chronic gastritis in outpatients of Peru. Observational, analytical, and cross-sectional study. Secondary data analysis was conducted. The dependent variable was chronic gastritis, obtained from patient references, and verified in the medical history according to the pathological history mentioned during medical consultation. The independent variable was the altitude of the residential areas (categorized into low altitude, intermediate altitude, high and very high), and the secondary co-variables were age, sex, and time living at altitude. Generalized linear models were used to estimate prevalence ratios using Poisson family and city as a cluster. Of the 4263 patients studied, 63% were female; the median age was 42 years. The overall prevalence of chronic gastritis was 12,9%. There was an association with chronic gastritis and altitude of residence at the intermediate and high levels, but not at the very high; with an adjusted prevalence ratio of 1.52 (95% confidence interval, 1.03 to 2.23); 2.01 (1.55 to 2.60) and 1.12 (0.84 to 1.48), respectively. We found a significant association between chronic gastritis and intermediate and high altitude but not at very high, which could be explained by hypobaric hypoxia in altitude that could lead to gastric wall lesions and other socio-demographic variables.


Temporal artery tap: usefulness and limitations in carotid sonography.
To examine the effectiveness of percussion of the superficial temporal artery for identification of the external carotid artery (ECA). The temporal artery tap maneuver was performed on 324 carotid arteries (163 patients). Evidence for transmission of the effect of the temporal tap was sought in the pulsed Doppler ultrasound waveforms of the ECA, common carotid artery (CCA), and internal carotid artery (ICA). The location and severity of stenotic lesions were recorded. The relative amplitudes of the oscillations created by the tap were compared. The temporal tap effect could be seen in 262 ECAs (81%), 174 CCAs (54%), and 106 ICAs (33%). The tap effect can be seen in the ICA at all grades of ICA disease. When the oscillations were seen in only one of the two major branches, that branch was always the ECA. When the temporal tap effect was found in the ICA, the amplitudes of the oscillations were the same as or greater than those of the ECA in 26% of cases. Waveform oscillations from the temporal tap maneuver often can be found beyond the ECA in the CCA and ICA. Thus, the temporal tap alone may not reliably distinguish the ECA from the ICA or CCA.


Effect of liners on cusp deflection and gap formation in composite restorations.
This study measured deformation of cusps and gap formation associated with MOD resin composite restorations in maxillary premolars with and without the use of low elastic modulus liners. Low elastic modulus liners may reduce the deformation by absorbing polymerization shrinkage stress. Forty maxillary premolars were mounted in stone and slot MOD cavities were prepared. Teeth were randomized into four groups. In Group A, cavities were etched, Single Bond was applied and the cavities were restored with Z-100 composite. In Group B, the same was done except that a layer of flowable composite (Revolution) was place and cured after the bonding agent. In Group C, the same steps were followed as Group A but a layer of glass ionomer (Vitrebond) was placed and cured before the bonding agent. In Group D, a thin layer of composite was placed (after the bonding agent) as a base and cured and the cavities were filled. The distance between indexed cusp tips was measured before the restorations were finished and five minutes and 24 hours after the restorations were completed. The samples were then sectioned mesiodistally and epoxy resin replicas were made and prepared for SEM evaluation of gap formation. The mean contraction of the cusps in microm at 5 minutes and 24 hours, respectively, for each group was A) 47 and 30, B) 35 and 21, C) 23 and 8 and D) 40 and 28. Groups A and D resulted in the highest deformation, B was intermediate and C was the lowest. There was no statistically significant difference in gap formation between the groups.


Expression and methylation of imprinted genes during in vitro differentiation of mouse parthenogenetic and androgenetic embryonic stem cell lines.
Messenger RNA and methylation levels of four imprinted genes, H19, Igf2r, Igf-2 and Snrpn were examined by northern and Southern blotting in mouse parthenogenetic, androgenetic and normal or wild-type embryonic stem cell lines during their differentiation in vitro as embryoid bodies. In most instances, mRNA levels in parthenogenetic and androgenetic embryoid bodies differed from wild type as expected from previously determined patterns of monoallelic expression in midgestation embryos and at later stages of development. These findings implicate aberrant mRNA levels of these genes in the abnormal development of parthenogenetic and androgenetic embryos and chimeras. Whereas complete silence of one of the parental alleles has previously been observed in vivo, we detected some mRNA in the corresponding embryonic stem cell line. This 'leakage' phenomenon could be explained by partial erasure, bypass or override of imprints, or could represent the actual activity status at very early stages of development. The mRNA levels of H19, Igf2r and Igf-2 and the degree of methylation at specific associated sequences were correlated according to previous studies in embryos, and thereby are consistent with suggestions that the methylation might play a role in controlling transcription of these genes. Paternal-specific methylation of the H19 promoter region is absent in sperm, yet we observed its presence in undifferentiated androgenetic embryonic stem cells, or before the potential expression phase of this gene in embryoid bodies. As such methylation is likely to invoke a repressive effect, this finding raises the possibility that it is part of the imprinting mechanism of H19, taking the form of a secondary imprint or postfertilization epigenetic modification necessary for repression of the paternal allele.


Generation of multilayered structures for biomedical applications using a novel tri-needle coaxial device and electrohydrodynamic flow.
In this short communication, we describe the scope and flexibility of using a novel device containing three coaxially arranged needles to form a variety of novel morphologies. Different combinations of materials are subjected to controlled flow through the device under the influence of an applied electric field. The resulting electrohydrodynamic flow allows us to prepare double-layered bubbles, porous encapsulated threads and nanocapsules containing three layers. The ability to process such multilayered structures is very significant for biomedical engineering applications, for example, generating capsules for drug delivery, which can provide multistage controlled release.


Application of a source apportionment model in consideration of volatile organic compounds in an urban stream.
Position-dependent concentrations of trichloroethylene and methyl-tert-butyl ether are considered for a 2.81-km section of the Aberjona River in Massachusetts, USA. This river flows through Woburn and Winchester (Massachusetts, USA), an area that is highly urbanized, has a long history of industrial activities dating to the early 1800s, and has gained national attention because of contamination from chlorinated solvent compounds in Woburn wells G and H. The river study section is in Winchester and begins approximately five stream kilometers downstream from the Woburn wells superfund site. Approximately 300 toxic release sites are documented in the watershed upstream from the terminus of the study section. The inflow to the river study section is considered one source of contamination. Other sources are the atmosphere, a tributary flow, and groundwater flows entering the river; the latter are categorized according to stream zone (1, 2, 3, etc.). Loss processes considered include outflows to groundwater and water-to-atmosphere transfer of volatile compounds. For both trichloroethylene and methyl-tert-butyl ether, degradation is neglected over the timescale of interest. Source apportionment fractions with assigned values alphainflow, alpha2, alpha3, etc. are tracked by a source apportionment model. The strengths of the groundwater and tributary sources serve as fitting parameters when minimizing a reduced least squares statistic between water concentrations measured during a synoptic study in July 2001 versus predictions from the model. The model fits provide strong evidence of substantial unknown groundwater sources of trichloroethylene and methyl-tert-butyl ether amounting to tens of grams per day of trichloroethylene and methyl-tert-butyl ether in the river along the study section. Modeling in a source apportionment manner can be useful to water quality managers allocating limited resources for remediation and source control.


Silk genes support the single origin of orb webs.
The orb web is a spectacular evolutionary innovation that enables spiders to catch flying prey. This elegant, geometric structure is woven with silk fibers that are renowned for their superior mechanical properties. We used silk gland expression libraries to address a long-standing controversy concerning the evolution of the orb-web architecture. Contrary to the view that the orb-web design evolved multiple times, we found that the distribution and phylogeny of silk proteins support a single, ancient origin of the orb web at least 136 million years ago. Furthermore, we substantially expanded the repository of silk sequences that can be used for the synthesis of high-performance biomaterials.


Development of a direct assay for alpha-amylase.
We describe a direct colorimetric assay for alpha-amylase, with 2-chloro-4-nitrophenyl-alpha-maltotrioside as substrate. Both human pancreatic and salivary amylase split this substrate without the use of helper enzymes, yielding free 2-chloro-4-nitrophenol, which is monitored at 405 nm. The performance of this reagent compares well with that of Pantrak Amylase (Behring Diagnostics) for both serum and urine samples. The reagent is very stable in dry powder form and is stable for one month at 2 to 8 degrees C after reconstitution. Because of the rapid color development and linear kinetics (less than 30 s), the assay is easily automated. Results can be obtained in less than 5 min.


Treatment of thyroid cancer.
The management of differentiated thyroid cancer remains controversial. A consecutive series of 234 thyroidectomies done by one surgeon in various Nairobi Hospitals from July 1990 to June 1993 were studied. 18 patients were operated on for thyroid cancer. The procedures ranged from lobectomy to total thyroidectomy. Young adults from 20-40 years of age were mainly affected, with a male/female sex ratio of 8:1. The immediate and subsequent morbidity of the operation of total thyroidectomy, in our opinion, overweighs its advantages of improved disease control. This is particularly so when low socio-economic standards lead to lack of compliance with replacement therapy.


Effect of ifosfamide treatment on glutathione and glutathione conjugation activity in patients with advanced cancers.
Several studies have suggested that the glutathione/glutathione S-transferase (GSH/GST) system is involved in resistance of tumors toward ifosfamide and other cytostatic agents. Besides, ifosfamide metabolites (in vitro) as well as ifosfamide treatment (in vivo) have been shown to decrease cellular GSH availability. In the present study, the in vivo effects of three different ifosfamide treatment schedules on the GSH/GST system were studied in patients with advanced cancers (n = 24): continuous i.v. infusions of 1300 mg/m2 daily for 10 days and 5000 mg/m2/day for 24 h, as well as a 4-h infusion of 3000 mg/m2 daily for 3 days. The GSH/GST system was characterized by administering bromisoval, a probe drug to assess GSH conjugation activity in vivo, as well as by daily monitoring of GSH concentrations in blood cells and plasma. Bromisoval pharmacokinetics was assessed before and at the end of the ifosfamide treatment. Blood cell GSH levels decreased significantly (P < 0.05) during the 3- and 10-day ifosfamide treatment schedules; the 24-h treatment had no effect. The ifosfamide treatment schedules had only minimal effects on bromisoval pharmacokinetics. Assuming that the kinetics of the probe drug provide an accurate reflection of enzyme activity, this suggests that GST activity remains unchanged. Because GSH conjugation of bromisoval enantiomers requires both GST activity and GSH availability, these results also indicate that, despite the 35% decrease in GSH in blood cells of two patient groups, the GSH availability of the cancer patients was not rate-limiting for GSH conjugation of bromisoval enantiomers. If GSH levels in blood cells reflect those in tumors/other tissues, the present results indicate that ifosfamide may be used clinically to decrease GSH levels. However, whether a 35% decrease is sufficient to increase tumor sensitivity toward (other) cytostatics remains uncertain.


Long-term survival of patients with breast cancer: a study of the curability of the disease.
A retrospective analysis was made of 3878 cases of breast carcinoma first seen in Edinburgh from 1954 to 1964. During this time there was a policy to treat breast cancer by simple mastectomy and x-ray therapy, and over 90% of cases classified as international stages I and II were so treated. The mortality in these women was compared with that in an equivalent normal population using Scottish national age-specific death rates. For every year of follow-up within 20 years of initial treatment there was an excess mortality from all causes. There was an overall excess mortality of 58% among patients with breast cancer 15-20 years after initial treatment, and 20 times more deaths occurred in this period from breast cancer than in a normal population. For patients disease-free after 15 years there was still a 28% excess mortality from all causes. Factors known to be of major prognostic significance for five-year survivorship had less influence than might have been expected when the ratio of observed to expected deaths was considered for longer periods of follow-up. The effect of clinical staging (I, II, or III), though initially marked, largely disappeared by the 10th year of follow-up, and after allowing for age there was no evidence beyond 10 years of an effect on survival of the original stage of the disease. Similarly, the effect of tumour size on survival disappeared after 10 years. Women who were premenopausal at presentation still had a significant excess of deaths in the fourth quinquennium of follow-up. In the menopausal and postmenopausal groups combined there was still a small non-significant excess of deaths from all causes after 15 years but this almost disappeared when patients who had already relapsed were excluded. In terms of overall mortality only patients who have undergone the menopause before presentation and who are disease-free 15 years after primary treatment may prove to be cured by conventional techniques such as simple mastectomy and postoperative radiotherapy.


Heterogeneous response of antimitochondrial autoantibodies and bile duct apical staining monoclonal antibodies to pyruvate dehydrogenase complex E2: the molecule versus the mimic.
The 2-oxo-acid dehydrogenase complexes and, in particular, the E2 component of the pyruvate dehydrogenase complex (PDC) are the target of antimitochondrial antibodies (AMA). More than 95% of primary biliary cirrhosis (PBC) patients have detectable levels of autoantibodies to PDC-E2 and in general these react with a region of the molecule that contains the prosthetic group lipoic acid (LA). LA is vital to the function of the enzyme, although there is conflicting evidence as to whether its presence is required for PDC-E2 recognition by AMA. Some, but not all, monoclonal antibodies (mAbs) to PDC-E2 produce an intense staining pattern at the apical surface of bile duct epithelial cells (BEC) in patients with PBC, and it has been argued that the molecule at the apical surface of PBC bile duct cells is a modified form of PDC-E2 or a cross-reactive molecule, acting as a molecular mimic. Herein, we characterize the epitopes recognized by 4 anti-PDC-E2 mAbs that give apical staining patterns (3 mouse and 1 human). In particular, by using a combination of recombinant antigens, competitive inhibition assays, and a unique peptide-on-bead assay, we determined that these apically staining mAbs recognize 3 or 4 distinct epitopes on PDC-E2. More importantly, this suggests that a portion spanning the entire inner lipoyl domain of PDC-E2 can be found at the BEC apical surface. In addition, competition assays with patient sera and a PDC-E2-specific mAb showed significant epitope overlap with only 1 of the 3 mouse mAbs and showed a differential response to the peptide bound to beads. These findings further highlight the heterogeneous response of patient autoantibodies to the inner lipoyl domain of PDC-E2.


Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study.
Targeting the endothelial adhesion molecules that control leukocyte trafficking into a tissue has been explored as a biological therapy for inflammatory diseases. However, these molecules also participate in leukocyte migration for immune surveillance, and inhibiting the physiological level of an adhesion molecule might promote infection or malignancy. We explored the concept of targeting endothelial adhesion molecule transcription during inflammation in a human system. Intercellular adhesion molecule 1 (ICAM-1) mediates leukocyte migration across the retinal endothelium in noninfectious posterior uveitis. We observed an increase in the transcription factor, nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NF-κB1), in parallel with ICAM-1, in human retinal endothelial cells treated with tumor necrosis factor-alpha (TNF-α), and identified putative binding sites for NF-κB1 within the ICAM-1 regulatory region. We targeted induced NF-κB1 expression in endothelial cells with small interfering (si)RNA. Knockdown of NF-κB1 significantly decreased cell surface expression of ICAM-1 protein induced by TNF-α but did not reduce constitutive ICAM-1 expression. Consistently, NF-κB1 knockdown significantly reduced leukocyte binding to cell monolayers in the presence of TNF-α but did not impact baseline binding. Findings of this proof-of-concept study indicate that induced transcription of endothelial adhesion molecules might be targeted therapeutically for inflammatory disease in humans.


Surgical care in the Solomon Islands: a road map for universal surgical care delivery.
Access to surgical care and emergency obstetrical care is limited in low-income countries. The Solomon Islands is one of the poorest countries in the Pacific region. Access to surgical care in Solomon Islands is limited and severely affected by a country made up of islands. Surgical care is centralized to the National Referral Hospital (NRH) on Guadalcanal, leaving a void of care in the provinces where more than 80% of the people live. To assess the ability to provide surgical care to the people living on outer islands in the Solomon Islands, the provincial hospitals were evaluated using the World Health Organization's Global Initiative for Emergency and Essential Surgical Care Needs Assessment Tool questionnaire. Data on infrastructure, workforce, and equipment available for treating surgical disease was collected at each provincial hospital visited. Surgical services are centralized to the NRH on Guadalcanal in Solomon Islands. Two provincial hospitals provide surgical care when a surgeon is available. Six of the hospitals evaluated provide only very basic surgical procedures. Infrastructure problems exist at every hospital including lack of running water, electricity, adequate diagnostic equipment, and surgical supplies. The number of surgeons and obstetricians employed by the Ministry of Health is currently inadequate for delivering care at the outer island hospitals. Shortages in the surgical workforce can be resolved in Solomon Islands with focused training of new graduates. Training surgeons locally, in the Pacific region, can minimize the "brain drain." Redistribution of surgeons and obstetricians to the provincial hospitals can be accomplished by creating supportive connections between these hospitals, the NRH, and international medical institutions.


Prevalence of Falls and Associated Factors among Community-Dwelling Older Adults: A Cross-Sectional Study.
As a person ages, the risk of falls increases, which affects quality of life and represents a financial burden to health- and social-systems, and a greater morbidity and mortality risk. Falls lead to decreased social contact, anxiety, long-term physical disability, severe dependency and hospitalizations. Currently, few studies address this phenomenon using a uniform methodology; therefore, this study aims to explore the prevalence of falls and associated-variables in older adults across Europe. In this cross-sectional analysis, we used data from Wave 6 of SHARE. The prevalence of falls was assessed through the answer "falling down" to the question "For the past six months at least, have you been bothered by any of the health conditions on this card?". Multilevel logistic regression was used, using falls as a dependent variable. Multilevel univariable logistic regression models were made to identify potential associated factors. From the 41,098 participants, 56.3% were female, and the average age was of 70.0 ± 8.9 years. The prevalence of falls was 8.2% (CI 8.0% to 8.4%), being higher in women (10.1% vs. 5.8%) and increasing with age. Age, female gender, being frail or pre-frail, higher scores on the EURO-D scale, polypharmacy and fear of falling were found to be significantly associated with falls. We found that falls are prevalent in the European community-dwelling population, with variations between countries. As a public health priority, identification of the variables associated with falls is important in order to identify/monitor the risk in older groups and develop tailored and cost-effective interventions for falls prevention.


Antipsychotics and the risk of diabetes and death among adults with serious mental illnesses.
Individuals with schizophrenia exposed to second-generation antipsychotics (SGA) have an increased risk for diabetes, with aripiprazole purportedly a safer drug. Less is known about the drugs' mortality risk or whether serious mental illness (SMI) diagnosis or race/ethnicity modify these effects. Authors created a retrospective cohort of non-elderly adults with SMI initiating monotherapy with an SGA (olanzapine, quetiapine, risperidone, and ziprasidone, aripiprazole) or haloperidol during 2008-2013. Three-year diabetes incidence or all-cause death risk differences were estimated between each drug and aripiprazole, the comparator, as well as effects within SMI diagnosis and race/ethnicity. Sensitivity analyses evaluated potential confounding by indication. 38 762 adults, 65% White and 55% with schizophrenia, initiated monotherapy, with haloperidol least (6%) and quetiapine most (26·5%) frequent. Three-year mortality was 5% and diabetes incidence 9.3%. Compared with aripiprazole, haloperidol and olanzapine reduced diabetes risk by 1.9 (95% CI 1.2-2.6) percentage points, or a 18.6 percentage point reduction relative to aripiprazole users' unadjusted risk (10.2%), with risperidone having a smaller advantage. Relative to aripiprazole users' unadjusted risk (3.4%), all antipsychotics increased mortality risk by 1.1-2.2 percentage points, representing 32.4-64.7 percentage point increases. Findings within diagnosis and race/ethnicity were generally consistent with overall findings. Only quetiapine's higher mortality risk held in sensitivity analyses. Haloperidol's, olanzapine's, and risperidone's lower diabetes risks relative to aripiprazole were not robust in sensitivity analyses but quetiapine's higher mortality risk proved robust. Findings expand the evidence on antipsychotics' risks, suggesting a need for caution in the use of quetiapine among individuals with SMI.


The responses of soil and rhizosphere respiration to simulated climatic changes vary by season.
Responses of soil respiration (Rs) to anthropogenic climate change will affect terrestrial carbon storage and, thus, feed back to warming. To provide insight into how warming and changes in precipitation regimes affect the rate and temperature sensitivity of Rs and rhizosphere respiration (Rr) across the year, we subjected a New England old-field ecosystem to four levels of warming and three levels of precipitation (ambient, drought, and wet treatments). We measured Rs and heterotrophic respiration (Rh) monthly (in areas of the plots with and without plants, respectively) and estimated Rr by calculating the difference in respiration between Rs and Rh. Even in this mesic ecosystem, Rs and Rr responded strongly to the precipitation treatments. Drought reduced Rs and Rr, both annually and during the growing season. Annual cumulative Rs responded nonlinearly to precipitation treatments; both drought and supplemental precipitation suppressed Rs compared to the ambient treatment. Warming increased Rs and Rr in spring and winter when soil moisture was optimal but decreased these rates in summer when moisture was limiting. Cumulative winter Rr increased by about 200% in the high warming (approximately 3.5 degrees C) treatment. The effect of climate treatments on the temperature sensitivity of Rs depended on the season. In the fall, the drought treatment decreased apparent Q10 relative to the other precipitation treatments. The responses of Rs to warming and altered precipitation were largely driven by changes in Rr. We emphasize the importance of incorporating realistic soil moisture responses into simulations of soil carbon fluxes; the long-term effects of warming on carbon--climate feedback will depend on future precipitation regimes. Our results highlight the nonlinear responses of soil respiration to soil moisture and, to our knowledge, quantify for the first time the loss of carbon through winter rhizosphere respiration due to warming. While this additional loss is small relative to the cumulative annual flux in this system, such increases in rhizosphere respiration during the non-growing season could have greater consequences in ecosystems where they offset or reduce subsequent warming-induced gains in plant growth.


The direct cost of "Thriasio" school screening program.
There is great diversity in the policies for scoliosis screening worldwide. The initial enthusiasm was succeeded by skepticism and the worth of screening programs has been challenged. The criticisms of school screening programs cite mainly the negative psychological impact on children and their families and the increased financial cost of visits and follow-up radiographs. The purpose of this report is to evaluate the direct cost of performing the school screening in a district hospital. A cost analysis was performed for the estimation of the direct cost of the "Thriasio" school-screening program between January 2000 and May 2006. The analysis involved all the 6470 pupils aged 6-18 years old who were screened at schools for spinal deformities during this period. The factors which were taken into consideration in order to calculate the direct cost of the screening program were a) the number of the examiners b) the working hours, c) the examiners' salary, d) the cost of transportation and finally e) the cost of examination per child. During the examined period 20 examiners were involved in the program and worked for 1949 working hours. The hourly salary for the trainee doctors was 6.80 euro, for the Health Visitors 6.70 euro and for the Physiotherapists 5.50 euro in current prices. The cost of transportation was 32 euro per year. The direct cost for the examination of each child for the above studied period was calculated to be 2.04 euro. The cost of our school-screening program is low. The present study provides a strong evidence for the continuation of the program when looking from a financial point of view.


Examination of molecular packing in orthogonal smectic liquid crystal phases: a guide for molecular design of functional smectic phases.
The reported layer spacings (dsmectic) of six homologues of mesogens exhibiting orthogonal smectic phases (SmE, SmB, and SmA phases) are reexamined. The slopes of the linear dependences on chain length (n, the number of carbon atoms in the hydrocarbon chain) are clearly categorized into two groups: 1.9 Å (CH2)-1 and 1.4 Å (CH2)-1. It is clarified that in the former the molecules take a rod-like form (rod-form; category-I), whereas in the latter the molecules are bent around the connection between the core and chain moieties (bent-form; category-II). The average relative positions of adjacent molecules within the smectic structures are deduced from the intercept of the linear functions of dsmectic against n. The relation between and the features of molecules belonging to the two categories are discussed for molecular design of functional smectic liquid crystals.


Impaired brainstem and thalamic high-frequency oscillatory EEG activity in migraine between attacks.
Introduction We investigated whether interictal thalamic dysfunction in migraine without aura (MO) patients is a primary determinant or the expression of its functional disconnection from proximal or distal areas along the somatosensory pathway. Methods Twenty MO patients and twenty healthy volunteers (HVs) underwent an electroencephalographic (EEG) recording during electrical stimulation of the median nerve at the wrist. We used the functional source separation algorithm to extract four functionally constrained nodes (brainstem, thalamus, primary sensory radial, and primary sensory motor tangential parietal sources) along the somatosensory pathway. Two digital filters (1-400 Hz and 450-750 Hz) were applied in order to extract low- (LFO) and high- frequency (HFO) oscillatory activity from the broadband signal. Results Compared to HVs, patients presented significantly lower brainstem (BS) and thalamic (Th) HFO activation bilaterally. No difference between the two cortical HFO as well as in LFO peak activations between the two groups was seen. The age of onset of the headache was positively correlated with HFO power in the right brainstem and thalamus. Conclusions This study provides evidence for complex dysfunction of brainstem and thalamocortical networks under the control of genetic factors that might act by modulating the severity of migraine phenotype.


Maternal and neonatal outcome of labour induction at term comparing two regimens of misoprostol.
To compare the efficacy and safety of two misoprostol dosing regimens for induction of labour in primiparous (1P) and multiparous (>1P) women. Retrospective study of induction of labour using vaginal misoprostol 25 μg vs. 50 μg every 6 h in 942 women at a tertiary centre. The main outcome variables are induction-to-delivery interval, latency period duration, vaginal delivery within 24 h, and maternal and foetal safety outcome. With the 50 μg regimen, induction-to-delivery intervals were significantly shorter: 18.4 h vs. 24.6 h (1P) and 14 h vs. 17.9 h (>1P), as was latency period duration (by 5.4 and 4 h, respectively). Vaginal delivery within 24 h was significantly more frequent, as were non-reassuring foetal heart rate (1P: 20% vs. 14%) and tachysystole (1P: 31% vs. 11%; >1P: 21% vs. 7%). No uterine rupture was reported. Neonatal outcomes were similar except for significantly more frequent infant referral to neonatal intensive care in the >1P group receiving the 50 μg regimen (11% vs. 4%). Vaginal misoprostol 25 μg seems to maintain efficacy with more acceptable maternal and neonatal safety. As induction of labour is an off-label use for misoprostol, safety should be prioritised with the lower dosage regimen despite the longer induction-to-delivery interval.


Localization of distinct reflections in rooms using spherical microphone array eigenbeam processing.
This paper presents an experimental and comparative study of several spherical microphone array eigenbeam (EB) processing techniques for localization of early reflections in room acoustic environments, which is a relevant research topic in both audio signal processing and room acoustics. This paper focuses on steered beamformer-based and subspace-based localization techniques implemented in the spherical EB domain, including the plane-wave decomposition, eigenbeam delay and sum, eigenbeam minimum variance distortionless response, eigenbeam multiple signal classification (EB-MUSIC), and eigenbeam estimation of signal parameters via rotational invariance techniques (EB-ESPRIT) methods. The directions of arrival of the original sound source and the associated reflection signals in acoustic environments are estimated from acoustic maps of the rooms, which are obtained using a spherical microphone array. The EB-domain-based frequency smoothing and white noise gain control techniques are derived and employed to improve the performance and robustness of reflection localization. The applicability of the presented methods in practice is confirmed by experiments carried out in real rooms.


Cerebral vasospasm following tumor resection.
To perform a systematic review of the literature of cases of vasospasm following tumor resection and to discuss possible etiologies, diagnosis and management of this complication. A literature search was performed using relevant search key words to identify cases of 'cerebral vasospasm' as a complication following tumor resection. Articles were identified by searching MEDLINE and PubMed databases. Relevant cross references were added by manually searching the references of all retrieved articles. There were a total of 56 patients in our initial search but only 40 cases matched our inclusion criteria. Pituitary tumors and sellar area were the most common pathologies and location associated with this complication. Average time interval to development of symptomatic vasospasm was 8 days and mortality was high (30%). The most common encountered causative factors were vascular factors (vessel encasement, displacement and narrowing) and presence of blood in the basal cisterns in postoperative scans. 'Cerebral vasospasm' after tumor resection is a rare but challenging complication with very high morbidity and mortality in reported cases. Vasospasm following brain tumor surgery shares some of the same clinical variables (time interval, causative factors, morbidity and death rates) of vasospasm after aneurysmal subarachnoid hemorrhage. A high index of suspicion is required for early diagnosis and prompt management which are key elements of final outcome.


Liraglutide and Cardiovascular Outcomes in Type 2 Diabetes.
The cardiovascular effect of liraglutide, a glucagon-like peptide 1 analogue, when added to standard care in patients with type 2 diabetes, remains unknown. In this double-blind trial, we randomly assigned patients with type 2 diabetes and high cardiovascular risk to receive liraglutide or placebo. The primary composite outcome in the time-to-event analysis was the first occurrence of death from cardiovascular causes, nonfatal myocardial infarction, or nonfatal stroke. The primary hypothesis was that liraglutide would be noninferior to placebo with regard to the primary outcome, with a margin of 1.30 for the upper boundary of the 95% confidence interval of the hazard ratio. No adjustments for multiplicity were performed for the prespecified exploratory outcomes. A total of 9340 patients underwent randomization. The median follow-up was 3.8 years. The primary outcome occurred in significantly fewer patients in the liraglutide group (608 of 4668 patients [13.0%]) than in the placebo group (694 of 4672 [14.9%]) (hazard ratio, 0.87; 95% confidence interval [CI], 0.78 to 0.97; P<0.001 for noninferiority; P=0.01 for superiority). Fewer patients died from cardiovascular causes in the liraglutide group (219 patients [4.7%]) than in the placebo group (278 [6.0%]) (hazard ratio, 0.78; 95% CI, 0.66 to 0.93; P=0.007). The rate of death from any cause was lower in the liraglutide group (381 patients [8.2%]) than in the placebo group (447 [9.6%]) (hazard ratio, 0.85; 95% CI, 0.74 to 0.97; P=0.02). The rates of nonfatal myocardial infarction, nonfatal stroke, and hospitalization for heart failure were nonsignificantly lower in the liraglutide group than in the placebo group. The most common adverse events leading to the discontinuation of liraglutide were gastrointestinal events. The incidence of pancreatitis was nonsignificantly lower in the liraglutide group than in the placebo group. In the time-to-event analysis, the rate of the first occurrence of death from cardiovascular causes, nonfatal myocardial infarction, or nonfatal stroke among patients with type 2 diabetes mellitus was lower with liraglutide than with placebo. (Funded by Novo Nordisk and the National Institutes of Health; LEADER ClinicalTrials.gov number, NCT01179048.).


The receptor for advanced glycation end products and risk of peripheral arterial disease, amputation or death in type 2 diabetes: a population-based cohort study.
Patients with type 2 diabetes have a high risk for early and extensive development of peripheral arterial disease (PAD) and this excess risk is not explained by increased burden of traditional atherosclerotic risk factors. Activation of the receptor for advanced glycation end products (RAGE) could be one additional mechanism for accelerated PAD and increased risk for amputation and death. We investigated the association between RAGE plasma components and the risk for PAD, amputation and death in patients with type 2 diabetes. We also estimated the rate of amputation-free survival and survival without PAD. We investigated if plasma levels of carboxymethyl-lysine, S100A12 and endosecretory RAGE (esRAGE) were associated with two endpoints: survival without development of PAD and survival without amputation in a 12 years prospective population-based cohort of 146 patients with type 2 diabetes, free from PAD at inclusion. Influence of baseline plasma levels of RAGE ligands (individually and combined by a RAGE-score) were evaluated for both endpoints in the Cox-regression analysis. 106 patients survived without amputation and 93 survived without signs of PAD during follow up. Higher levels of S100A12 and RAGE-score were associated with increased risk for amputation or death, hazard ratios (HR) 1.29; 95% confidence interval (CI) [1.04, 1.59] and 1.79; 95% CI [1.07, 2.99] and with increased risk for PAD or death, HR 1.22; 95% CI [1.00, 1.49] and 1.56; [1.00, 2.44] after adjustment for age and sex. The effect was decreased after adjustment for Framingham cardiovascular disease score: risk for amputation or death, HR 1.17; 95% CI [0.94, 1.46] and 1.54; [0.95, 2.49], and risk for PAD or death, HR 1.12; 95% CI [0.91, 1.38] and 1.38; [0.91, 2.11] for S100A12 and RAGE-score respectively. The incidence for amputation or death was 2.8 per 100 person-years; 95% CI [2.0, 3.7] and the incidence rate for PAD or death was 3.6 per 100 person-years; 95% CI [2.7, 4.8]. Higher plasma levels of S100A12 and the combined effect (RAGE-score) of esRAGE, carboxymethyl-lysine and S100A12 seem to be associated with shorter PAD- and amputation-free survival in patients with type 2 diabetes. This may indicate a role for S100A12 in PAD by activation of the RAGE system.


Catecholate siderophores protect bacteria from pyochelin toxicity.
Bacteria produce small molecule iron chelators, known as siderophores, to facilitate the acquisition of iron from the environment. The synthesis of more than one siderophore and the production of multiple siderophore uptake systems by a single bacterial species are common place. The selective advantages conferred by the multiplicity of siderophore synthesis remains poorly understood. However, there is growing evidence suggesting that siderophores may have other physiological roles besides their involvement in iron acquisition. Here we provide the first report that pyochelin displays antibiotic activity against some bacterial strains. Observation of differential sensitivity to pyochelin against a panel of bacteria provided the first indications that catecholate siderophores, produced by some bacteria, may have roles other than iron acquisition. A pattern emerged where only those strains able to make catecholate-type siderophores were resistant to pyochelin. We were able to associate pyochelin resistance to catecholate production by showing that pyochelin-resistant Escherichia coli became sensitive when biosynthesis of its catecholate siderophore enterobactin was impaired. As expected, supplementation with enterobactin conferred pyochelin resistance to the entE mutant. We observed that pyochelin-induced growth inhibition was independent of iron availability and was prevented by addition of the reducing agent ascorbic acid or by anaerobic incubation. Addition of pyochelin to E. coli increased the levels of reactive oxygen species (ROS) while addition of ascorbic acid or enterobactin reduced them. In contrast, addition of the carboxylate-type siderophore, citrate, did not prevent pyochelin-induced ROS increases and their associated toxicity. We have shown that the catecholate siderophore enterobactin protects E. coli against the toxic effects of pyochelin by reducing ROS. Thus, it appears that catecholate siderophores can behave as protectors of oxidative stress. These results support the idea that siderophores can have physiological roles aside from those in iron acquisition.


Color constancy influenced by unnatural spatial structure.
The recognition of spatial structures is important for color constancy because we cannot identify an object's color under different illuminations without knowing which space it is in and how that space is illuminated. To show the importance of the natural structure of environments on color constancy, we investigated the way in which color appearance was affected by unnatural viewing conditions in which a spatial structure was distorted. Observers judged the color of a test patch placed in the center of a small room illuminated by white or reddish lights, as well as two rooms illuminated by white and reddish light, respectively. In the natural viewing condition, an observer saw the room(s) through a viewing window, whereas in an unnatural viewing condition, the scene structure was scrambled by a kaleidoscope-type viewing box. Results of single room condition with one illuminant color showed little difference in color constancy between the two viewing conditions. However, it decreased in the two-rooms condition with a more complex arrangement of space and illumination. The patch's appearance under the unnatural viewing condition was more influenced by simultaneous contrast than its appearance under the natural viewing condition. It also appears that color appearance under white illumination is more stable compared to that under reddish illumination. These findings suggest that natural spatial structure plays an important role for color constancy in a complex environment.


Copper Influences the Antibacterial Outcomes of a β-Lactamase-Activated Prochelator against Drug-Resistant Bacteria.
The unabated rise in bacterial resistance to conventional antibiotics, coupled with collateral damage to normal flora incurred by overuse of broad-spectrum antibiotics, necessitates the development of new antimicrobials targeted against pathogenic organisms. Here, we explore the antibacterial outcomes and mode of action of a prochelator that exploits the production of β-lactamase enzymes by drug-resistant bacteria to convert a nontoxic compound into a metal-binding antimicrobial agent directly within the microenvironment of pathogenic organisms. Compound PcephPT (phenylacetamido-cephem-pyrithione) contains a cephalosporin core linked to 2-mercaptopyridine N-oxide (pyrithione) via one of its metal-chelating atoms, which minimizes its preactivation interaction with metal ions and its cytotoxicity. Spectroscopic and chromatographic assays indicate that PcephPT releases pyrithione in the presence of β-lactamase-producing bacteria. The prochelator shows enhanced antibacterial activity against strains expressing β-lactamases, with bactericidal efficacy improved by the presence of low-micromolar copper in the growth medium. Metal analysis shows that cell-associated copper accumulation by the prochelator is significantly lower than that induced by pyrithione itself, suggesting that the location of pyrithione release influences biological outcomes. Low-micromolar (4-8 μg/mL) minimum inhibitory concentration (MIC) values of PcephPT in ceftriaxone-resistant bacteria compared with median lethal dose (LD50) values greater than 250 μM in mammalian cells suggests favorable selectivity. Further investigation into the mechanisms of prochelators will provide insight for the design of new antibacterial agents that manipulate cellular metallobiology as a strategy against infection.


LEDGF dominant interference proteins demonstrate prenuclear exposure of HIV-1 integrase and synergize with LEDGF depletion to destroy viral infectivity.
Target cell overexpression of the integrase binding domain (IBD) of LEDGF/p75 (LEDGF) inhibits HIV-1 replication. The mechanism and protein structure requirements for this dominant interference are unclear. More generally, how and when HIV-1 uncoating occurs postentry is poorly defined, and it is unknown whether integrase within the evolving viral core becomes accessible to cellular proteins prior to nuclear entry. We used LEDGF dominant interference to address the latter question while characterizing determinants of IBD antiviral activity. Fusions of green fluorescent protein (GFP) with multiple C-terminal segments of LEDGF inhibited HIV-1 replication substantially, but minimal chimeras of either polarity (GFP-IBD or IBD-GFP) were most effective. Combining GFP-IBD expression with LEDGF depletion was profoundly antiviral. CD4(+) T cell lines were rendered virtually uninfectable, with single-cycle HIV-1 infectivity reduced 4 logs and high-input (multiplicity of infection = 5.0) replication completely blocked. We restricted GFP-IBD to specific intracellular locations and found that antiviral activity was preserved when the protein was confined to the cytoplasm or directed to the nuclear envelope. The life cycle block triggered by the cytoplasm-restricted protein manifested after nuclear entry, at the level of integration. We conclude that integrase within the viral core becomes accessible to host cell protein interaction in the cytoplasm. LEDGF dominant interference and depletion impair HIV-1 integration at distinct postentry stages. GFP-IBD may trigger premature or improper integrase oligomerization.


Raspberry anthocyanin consumption prevents diet-induced obesity by alleviating oxidative stress and modulating hepatic lipid metabolism.
Evidence indicates that raspberries have beneficial effects on chronic diseases. The objective of this study was to examine the beneficial effects of raspberry anthocyanin (RA) on high fat diet-induced obesity and investigate the underlying molecular mechanism. C57BL/6 mice were administered a low-fat diet, high-fat diet, and high-fat diet supplemented with RA at a dose of 200 mg kg-1 of food for 12 weeks. It was found that RA reduced the body weight gain by 63.7%. Furthermore, RA significantly elevated serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities and fecal butyric acid level, remarkably reduced the serum and hepatic lipid profiles, and markedly down-regulated the expression of the tumor necrosis factor α (TNFα), interleukin-6 (IL-6), and nuclear factor κB (NF-κB) genes. Metabolomics analysis conducted using gas chromatography time-of-flight mass spectrometry (GC-TOF/MS) indicated that RA administration promoted the recovery of metabolites involved in glycerophospholipid metabolism, insulin signaling pathway, and glutathione metabolism in the livers of obese mice. These findings suggest that RA may ameliorate diet-induced obesity by alleviating oxidative stress and modulating lipid metabolism.


Improving the effectiveness of smoking cessation in primary care: lessons learned.
The 'Smokescreen' smoking cessation programme was introduced in Christchurch in 1995, with an initial study showing six-month, self-reported quit rates of 10% and 17% (with a validated deception rate) in primary and secondary care settings. Substantial modifications were made to try to improve this rate in the primary care setting and the programme has been implemented widely. Our primary aim was to estimate programme utilisation and six-month quit rates for enrolled patients in this general primary care setting. We also aimed to use a wide range of patient, practice and environmental variables to estimate any predictive effect on outcome. Prospective longitudinal cohort study. The nicotine replacement therapy (NRT) -based programme was implemented by Pegasus Health, an independent practitioner association (IPA) situated in the Christchurch urban area, to which the majority of Christchurch-based GPs belong. A cohort of 516 patients enrolling in the programme over a two-month period were contacted six months after their nominated quit date. The main outcome measure was the six-month, self-reported quit rate. Of the 516 participants, 334 (65%) were contacted by mail or telephone. The overall six-month quit rate was 36% (95% Confidence Interval (CI) 31-41). Univariate analysis initially showed duration of NRT (p = 0.03) and age band (p = 0.004) were significant predictors of quitting, while living with a smoker (p = 0.02), having made no previous quit attempts (p = 0.02) and having heart disease (p = 0.01) were all significant predictors of continued smoking at six months. Factors that did not predict whether respondents were smoking at six months included previous use of NRT, sex, ethnicity, who delivered the intervention, years of smoking, cigarette dose, and NZDep96 score. However, there was interaction between these factors as after multivariate analysis the only significant predictors of outcome were having others living in the house who smoked (odds ratio (OR) 0.55, 95% CI 0.33--0.93, p = 0.03) and having made no previous quit attempts (OR 0.29, 95% CI 0.12-0.71, p = 0.02). Both these factors were significantly associated with continuing to smoke. This programme compares favourably with six-month quit rates for NRT-based programmes reported in the international literature of 14-22%. The effectiveness of an NRT-based smoking cessation programme in a general primary care setting appears to have been significantly enhanced by local adaptation, the flexibility of a primary-care-team approach and subsidisation of NRT, together with facilitation responsive to individual practice needs. The success of this programme in helping individual patients quit, as well as its successful implementation in a wide primary care setting, suggests General Practice can play an important role in smoking cessation in a country with a high burden of disease from smoking-related illnesses. The programme is congruent with the current, national, smoking cessation guidelines endorsed by the RNZCGP. Widespread adoption of this kind of model in IPA/primary health organisation (PHO) settings throughout New Zealand should be encouraged and supported.


Stressing out in medieval Denmark: An investigation of dental enamel defects and age at death in two medieval Danish cemeteries.
The influence of early life stress on later life experiences has become a major focus of research in medicine and more recently in bioarchaeology. Dental enamel, which preserves a record of childhood stress events, represents an important resource for this investigation when paired with the information from adult skeletal remains, such as age at death. The purpose of this research was to use a life history approach to the exploration of sex differences in the relationship between childhood stress and adult longevity by examining accentuated striae of Retzius (AS). A medieval Danish sample (n=70) drawn from the rural cemetery of Sejet and the urban cemetery of Ole Wormsgade was considered for AS and age at death. The results suggest sex differences in survivorship, with more stress being associated with reduced survivorship in males and increased survivorship in females. A consideration of AS formation time also suggests a difference in the impact of developmental timing between males and females. These results are interpreted in terms of differential frailty and selective mortality, drawing in both biomedical and cultural perspectives.


Proposed pathogenesis of idiopathic loin pain-hematuria syndrome.
To study loin pain-hematuria syndrome (LPHS) pathogenesis, we evaluated 43 consecutive patients for whom urological evaluation failed to disclose the cause of their recurrent flank pain and hematuria. Each underwent percutaneous kidney biopsy. In 9 patients, the biopsy specimen showed immunoglobulin A nephritis, an established cause of LPHS. We suggest these cases be designated secondary LPHS. They are not included in this analysis. The remaining patients (N = 34) are designated idiopathic (primary) LPHS. They are the basis of this report. Demographics of patients with primary LPHS are mean age of 30.8 +/- 10.3 years; 74% women; 94% white; and history of kidney stones, 47%, although none was obstructing. Primary LPHS kidney biopsy specimens showed red blood cells (RBCs) in multiple tubules, consistent with glomerular hematuria. Glomeruli were normal by means of light and immunofluorescent microscopy; however, more than 50% of biopsy specimens showed unusually thin or thick glomerular basement membranes. To assess whether the biopsy itself caused RBCs in tubules, we compared RBCs in renal tubular cross-sections from primary LPHS biopsies with those of normal kidneys (donors, n = 10). The mean percentage of tubular cross-sections containing RBCs was greater in primary LPHS than normal specimens (7.2% +/- 6.5% versus 1.6% +/- 1.0% [SD]; P < 0.0001), confirming glomerular hematuria in patients with primary LPHS. Primary LPHS pathogenesis includes glomerular hematuria, apparently from structurally abnormal glomerular basement membrane. Primary LPHS pain may be initiated by obstructing RBC casts and perhaps microcrystals in those with a history of urolithiasis. Nevertheless, other factors are needed to explain the severe pain in patients with primary LPHS.


Effects of a manually assisted mechanical force on cutaneous temperature.
Digitized infrared segmental thermometry (DIST) is a tool used for measuring cutaneous temperature (CT). This project ascertains the effect of a manually assisted mechanical force producing a chiropractic adjustment in the lumbar spine after the Activator Methods Chiropractic Technique on CT during 2 different time recording periods (TRPs). Sixty-six healthy subjects (36 women and 30 men) without acute low back conditions or symptoms were recruited. Subjects were randomly divided into 2 groups based on the length of the acclimatization period (8 or 30 minutes; TRP(8) and TRP(30), respectively). In turn, each recording period group was divided into 3 subgroups (n = 11 per subgroup): treatment, sham, and control subgroups. Bilateral DIST was conducted at L-4 (TRP(30)) and L-5 (TRP(8)) using infrared cameras (Subluxation Station Insight 7000; Chiropractic Leadership Alliance, Mahwah, NJ). Before treatment (t(-0.5)), the TRP(8) CT was significantly different between the ipsilateral and the contralateral sides for all subgroups. At 10 minutes (t(10)) after intervention, CT increased significantly (P < .05) for the treatment group but not for the sham and control groups. In contrast, there were no significant differences in the TRP(30) CT before treatment between the ipsilateral and the contralateral sides; but at t(10), CT was significantly (P < .05) greater for all 3 subgroups compared with preintervention CT. Contacting the skin with the instrument with (treatment group TRP(30)) or without (sham group TRP(30)) a thrust with a sustained pressure stronger than the loading principle taught in the Activator Methods Chiropractic Technique protocol or a thrust respecting the standard loading principle (treatment group TRP(8)) of the instrument produced a CT cooling immediately after the adjustment. Furthermore, we observed that when contacting the skin with the instrument with a thrust respecting the standard loading principle (treatment group TRP(8)) of the instrument, it produced a secondary cooling at t(5) followed by a rewarming at t(10). Finally, contacting the skin with the instrument without a thrust and respecting the standard loading principle (sham TRP(8)) of the instrument did not produce a CT change.


Synthesis and characterization of core-shell star copolymers for in vivo PET imaging applications.
The synthesis of core-shell star copolymers via living free radical polymerization provides a convenient route to three-dimensional nanostructures having a poly(ethylene glycol) outer shell, a hydrophilic inner shell bearing reactive functional groups, and a central hydrophobic core. By starting with well-defined linear diblock copolymers, the thickness of each layer, overall size/molecular weight, and the number of internal reactive functional groups can be controlled accurately, permitting detailed structure/performance information to be obtained. Functionalization of these polymeric nanoparticles with a DOTA-ligand capable of chelating radioactive (64)Cu nuclei enabled the biodistribution and in vivo positron emission tomography (PET) imaging of these materials to be studied and correlated directly to the initial structure. Results indicate that nanoparticles with increasing PEG shell thickness show increased blood circulation and low accumulation in excretory organs, suggesting application as in vivo carriers for imaging, targeting, and therapeutic groups.


The bacterial genus Collimonas: mycophagy, weathering and other adaptive solutions to life in oligotrophic soil environments.
This minireview provides a synopsis of past and present research on the biology and ecology of members of the bacterial genus Collimonas. From the distribution, abundance and functional behaviours of these so-called collimonads emerges a general picture of bacterial adaptation to low-nutrient soil environments. Among these adaptations is the ability to extract nutrients from living fungi (mycophagy) and from rocks and minerals (weathering). This unique combination of properties will be discussed in the context of other interactions that collimonads have with their biotic and abiotic surroundings, such as the ability to inhibit fungal growth (fungistasis), protect plant roots from fungal disease (biocontrol), and degrade natural polymers and synthetic pollutants (biodegradation). Future research on Collimonas is expected to take advantage of the genomic tools and resources that are becoming available to uncover and describe the genes and gene functions that distinguish this group of bacteria and are the basis for its phenotypes. Potential applications of collimonads include the control of unwanted fungi, for example in agriculture, their use as biological indicators of soil quality and fertility, and as a source of bioactive compounds.


Providing feedback following Leadership WalkRounds is associated with better patient safety culture, higher employee engagement and lower burnout.
There is a poorly understood relationship between Leadership WalkRounds (WR) and domains such as safety culture, employee engagement, burnout and work-life balance. This cross-sectional survey study evaluated associations between receiving feedback about actions taken as a result of WR and healthcare worker assessments of patient safety culture, employee engagement, burnout and work-life balance, across 829 work settings. 16 797 of 23 853 administered surveys were returned (70.4%). 5497 (32.7% of total) reported that they had participated in WR, and 4074 (24.3%) reported that they participated in WR with feedback. Work settings reporting more WR with feedback had substantially higher safety culture domain scores (first vs fourth quartile Cohen's d range: 0.34-0.84; % increase range: 15-27) and significantly higher engagement scores for four of its six domains (first vs fourth quartile Cohen's d range: 0.02-0.76; % increase range: 0.48-0.70). This WR study of patient safety and organisational outcomes tested relationships with a comprehensive set of safety culture and engagement metrics in the largest sample of hospitals and respondents to date. Beyond measuring simply whether WRs occur, we examine WR with feedback, as WR being done well. We suggest that when WRs are conducted, acted on, and the results are fed back to those involved, the work setting is a better place to deliver and receive care as assessed across a broad range of metrics, including teamwork, safety, leadership, growth opportunities, participation in decision-making and the emotional exhaustion component of burnout. Whether WR with feedback is a manifestation of better norms, or a cause of these norms, is unknown, but the link is demonstrably potent.


Molecular cloning of cDNA for equine ovarian inhibin/activin beta A subunit.
cDNAs encoding equine inhibin/activin beta A subunit were isolated from an equine follicle cDNA library and characterized. Using primers based on the rat inhibin/activin beta A subunit cDNA sequence, a RT-PCR was performed to generate the probe for screening. Four positive clones were isolated. Analysis of the nucleotide sequence of these clones revealed that two pairs of identical clones were present, Eq-beta A-1 (0.9 kb) and Eq-beta A-2 (1.5 kb). Eq-beta A-2 clone contained a complete open reading frame encoding 426 amino acids. The deduced amino acid sequence of equine inhibin/activin beta A subunit showed high similarity (> 90%) to those of five other mammalian species. Northern blot analysis revealed that placenta from mare on day 180 of pregnancy contained a 1.5 kb inhibin/activin beta A subunit mRNA.


HIV-1 infects macrophages by exploiting an endocytic route dependent on dynamin, Rac1 and Pak1.
Recent studies provide compelling evidence that HIV-1 entry in cell lines and lymphocytes proceeds by endocytosis, but these studies are still lacking in macrophages, an important natural target cell for HIV-1. Macrophages exhibit continual and extensive endocytic activity as part of their natural functions, so we investigated the uptake pathways involved in productive HIV-1 entry. We find that caveolae are not utilised by HIV-1, because the main structural proteins, caveolin-1 and 2 are absent from most human leukocytes. We then focused on macropinocytosis; we find that HIV-1 entry into macrophages is sensitive to inhibitors of Na(+)/H(+) exchange, actin rearrangement, dynamin, Rho family GTPases, and Pak1, but not to inhibitors of PI-3 kinase and myosin II. This leads us to conclude that HIV entry into macrophages proceeds by an endocytic pathway that is not classical macropinocytosis. Because of the limitations of a purely pharmacological study such as this, the final elucidation of this pathway awaits the development of reliable forward genetic approaches in authentic macrophages.


Body mass index does not affect postoperative morbidity and oncologic outcomes of total mesorectal excision for rectal adenocarcinoma.
Obesity adds to the technical difficulty of colorectal surgery and is a risk factor for postoperative complications. We hypothesized that obese patients have increased morbidity and poor oncologic outcomes after proctectomy for rectal adenocarcinoma. Adult patients undergoing total mesorectal excision (TME) for rectal adenocarcinoma at a tertiary referral center were retrospectively identified from a prospectively maintained database. Operative characteristics, postoperative complication rates, and oncologic outcomes were compared in patients with BMI > or = 30 kg/m(2) and BMI < 30 kg/m(2). Between 1997 and 2009, 254 patients underwent proctectomy for rectal adenocarcinoma, of whom 27% were obese. There were no significant differences in demographics, comorbidities or preoperative oncologic characteristics between obese and nonobese groups. Patients with BMI > or = 30 kg/m(2) had longer operative times (p = 0.04) and higher intraoperative blood loss (p < 0.001) but comparable postoperative complication rates (p = 0.80), number of lymph nodes retrieved (p = 0.57), margin-negative resections (p = 0.44), and disease-free survival (p = 0.11). Obese patients had longer overall survival (p = 0.05). Tumor stage was the only variable associated with disease-free (p < 0.001) and overall survival (p < 0.001). Despite increased technical difficulty of resection, obesity does not increase the risk of postoperative morbidity or adversely affect oncologic outcomes after total mesorectal excision of rectal adenocarcinoma.


Ameliorative Effects of Hydromethanolic Extract of Citrullus lanatus (Watermelon) Rind on Semen Parameters, Reproductive Hormones and Testicular Oxidative Status Following Nicotine Administration in Male Wistar Rats.
The present study examines the possible ameliorative effects of the hydromethanolic extract of Citrullus lanatus rind (HECL) on some reproductive function and oxidative indices of the testes in male Wistar rats following administration of nicotine. Twenty male rats were assigned into four groups: Group A to D of five rats each. Group A served as control and received 2ml/kg body weight of 10% extract vehicle; Group B received 1mg/kg body weight of nicotine; Group C were co-administered 1mg/kg body weight nicotine and 500 mg/kg body weight of HECL and Group D received only 500mg/kg body weight of HECL. The drugs and extracts were administered orally to the rats for 42days; blood samples were collected by direct cardiac puncture for determination of serum concentrations of testosterone, Follicle Stimulating Hormone and Luteinizing Hormone. The testes were also harvested for determination of semen parameters: motility, morphology, viability and count and testicular tissue processed for superoxide dismutase and malondialdehyde concentration. Compared to Group A control rats, administration of HECL significantly increased sperm count and reproductive hormone concentrations amongst Group B rats (p<0.05). Treatment with nicotine caused a significant reduction in the levels of all reproductive hormones with significant diminution of some sperm parameters: motility, morphology and viability; and decrease in superoxide dismutase and increase in malondialdehyde concentration amongst Group B rats compared to Group A control rats (p<0.05). Co-administration of HECL with nicotine to Group C rats apparently reversed the effects of nicotine resulting in significant increases in sperm count and the reproductive hormones concentration as compared to Group A control rats (p<0.05). Amongst Group D rats, the extract also caused a significant increase in superoxide dismutase concentration and a significant decrease in malondialdehyde concentration compared with the Group A control rats (p<0.05). The findings suggest that the hydromethanolic extract of Citrullus lanatus rind possibly ameliorates the deleterious effects of nicotine on some reproductive indices in male Wistar rats.


The Role of Virtual Reality as a Psychological Intervention for Mental Health Disturbances during the COVID-19 Pandemic: A Narrative Review.
The COVID-19 pandemic spread throughout the world and created many problems. The COVID-19 pandemic caused an increase in mortality and morbidity, including mental health problems. Around the world, the movement control order (MCO) was strictly enforced, but the spread of the infection epidemic was still rampant. The magnitude of the increase in mental health illnesses has caused many individuals to suffer. Given that face-to-face interventions are challenging to carry out during an outbreak, we need to address this critical problem through an online approach, such as virtual reality (VR). This approach is vital to helping patients deal with their existing problems in more pragmatic, practical, and customer-friendly ways. Thus, in the present review, we proposed the development of a virtual digital device for this noble purpose. Various challenges, improvements, and expectations for VR applications were outlined and discussed in this narrative review.


Neural correlates of texture perception during active touch.
Previous studies have shown attenuation of cortical oscillations over bilateral sensorimotor cortex areas during passive perception of smooth textures applied to the skin. However, humans typically explore surfaces using dynamic hand movements. As movements may both modulate texture-related cortical activity and induce movement-related cortical activation, data from passive texture perception cannot be extrapolated to active texture perception. In the present study, we used electroencephalography to investigate cortical oscillatory changes during texture perception throughout active touch exploration. Three natural textured stimuli were selected: smooth silk, soft brushed cotton, and rough hessian. Texture samples were mounted on a purpose-built touch sensor which measured the load and position of the index finger, whilst electroencephalography from 129 channels recorded oscillatory brain activity. The data were fused to investigate oscillatory changes relating to active touch. Changes in oscillatory band power, event-related desynchronisation/synchronisation (ERD/ERS), were investigated in alpha (8-12 Hz) and beta (16-24 Hz) frequency bands. Active texture exploration revealed bilateral activation patterns over sensorimotor cortical areas. Beta-band ERD increased over contralateral sensorimotor regions for soft and smooth textures, and over ipsilateral sensorimotor areas for the smoothest texture. Analysis of covariance revealed that individual differences in perception of softness and smoothness were related to variations in cortical oscillatory activity. Differences may be due to increased high frequency vibrations for smooth and soft textures compared to rough. For the first time, active touch was quantified and fused with electroencephalography data streams, contributing to the understanding of the neural correlates of texture perception during active touch.


Improving HIV proteome annotation: new features of BioAfrica HIV Proteomics Resource.
The Human Immunodeficiency Virus (HIV) is one of the pathogens that cause the greatest global concern, with approximately 35 million people currently infected with HIV. Extensive HIV research has been performed, generating a large amount of HIV and host genomic data. However, no effective vaccine that protects the host from HIV infection is available and HIV is still spreading at an alarming rate, despite effective antiretroviral (ARV) treatment. In order to develop effective therapies, we need to expand our knowledge of the interaction between HIV and host proteins. In contrast to virus proteins, which often rapidly evolve drug resistance mutations, the host proteins are essentially invariant within all humans. Thus, if we can identify the host proteins needed for virus replication, such as those involved in transporting viral proteins to the cell surface, we have a chance of interrupting viral replication. There is no proteome resource that summarizes this interaction, making research on this subject a difficult enterprise. In order to fill this gap in knowledge, we curated a resource presents detailed annotation on the interaction between the HIV proteome and host proteins. Our resource was produced in collaboration with ViralZone and used manual curation techniques developed by UniProtKB/Swiss-Prot. Our new website also used previous annotations of the BioAfrica HIV-1 Proteome Resource, which has been accessed by approximately 10 000 unique users a year since its inception in 2005. The novel features include a dedicated new page for each HIV protein, a graphic display of its function and a section on its interaction with host proteins. Our new webpages also add information on the genomic location of each HIV protein and the position of ARV drug resistance mutations. Our improved BioAfrica HIV-1 Proteome Resource fills a gap in the current knowledge of biocuration.Database URL:http://www.bioafrica.net/proteomics/HIVproteome.html.


Activation of monocyte effector genes and STAT family transcription factors by inflammatory synovial fluid is independent of interferon gamma.
Activated monocytes play an important role in the pathogenesis of inflammatory arthritis. Blood monocytes which enter the inflamed joint become activated upon adherence to extracellular matrix and exposure to a complex inflammatory environment. We have analyzed the mechanism of monocyte activation by soluble factors present in inflammatory synovial fluid (SF). Greater than 75% of inflammatory SFs tested (a total of 22 fluids to date) increased cell surface expression and dramatically increased mRNA levels of monocyte activation markers Fc gamma RI, Fc gamma RIII, and HLA-DRA. This induction was not triggered by adherence, a known activating stimulus, and several lines of evidence showed that induction was not dependent upon interferon gamma (IFN-gamma). Induction was not prevented by neutralizing anti-IFN-gamma antibodies and IFN-gamma was not detected in the SFs using a sensitive enzyme-linked immunosorbent assay. The SFs also were not able to activate the IFN-gamma-activated transcription factor Stat1, thus providing further support for the absence of IFN-gamma. SFs did activate a related signal transducer and activator of transcription (STAT) family factor, termed Stat-SF, which bound specifically to the IFN-gamma response region (GRR), a well-characterized transcription element in the Fc gamma RI promoter. Based upon DNA-binding specificity and mobilities in gel shift assays, and reactivity with specific antisera, Stat-SF likely contains Stat3, or a closely related STAT family member. Neutralization of interleukin 6, a cytokine present in SFs which is known to activate Stat3, abolished the activation of Stat-SF and inhibited the induction of Fc gamma RI expression by SFs. These results demonstrate the activation of monocytes by inflammatory SF and suggest that monocyte activation at an inflammatory site may occur in the absence of IFN-gamma through the triggering of signal transduction pathways that activate STAT transcription factors.


Computed tomography urography for diagnosing bladder cancer.
To evaluate the use of computed tomography urography (CTU) for diagnosing bladder tumours in patients with macroscopic haematuria and aged >40 years. In all, 200 consecutive patients attending a fast-track haematuria clinic were assessed using 'same-day' CTU and flexible cystoscopy. Patients were aged >40 years and had macroscopic haematuria with no urine infection. CTU studies were reported by one uroradiologist and scored on a 3-point scale to quantify the probability of bladder cancer. All flexible cystoscopies were performed by the same cystoscopist with no knowledge of the findings of CTU, and scored using a 3-point scale. Cystoscopy, pathological findings and CTU were then compared. The prevalence of bladder tumours was 24%; when CTU was compared with the histopathological findings, there was one false-positive and three false-negative diagnoses, indicating a sensitivity of 0.93 and a specificity of 0.99, with a 0.98 positive and 0.97 negative predictive value for detecting bladder cancer. A review of the three false-negative cases showed that one was missed on original CTU reporting, the second had the appearance of prostate cancer on CTU and the third was a squamous metaplasia. CTU is an accurate method of detecting bladder tumours in the present patients, and is reliable and accurate for assessing the bladder. Our results support the use of CTU as a first-line screening tool for this high-risk group, the use of which will obviate the need for flexible cystoscopy in patients with a negative CTU and allow those with an obvious tumour to be referred directly for rigid cystoscopy and resection. The remaining patients should be referred for flexible cystoscopy. Such a pathway would accelerate patient assessment by using fewer tests and provide a true 'one-stop' clinic, allowing a comprehensive evaluation with a single test for the upper and lower urinary tract.


Training in hypoxia: modulation of metabolic and cardiovascular risk factors in men.
This study was designed to determine changes in metabolic and cardiovascular risk factors following normobaric hypoxic exercise training in healthy men. Following a randomized baseline maximal exercise test in hypoxia and/or normoxia, 34 physically active subjects were randomly assigned to either a normoxic (N = 14) or a hypoxic (N = 18) training group. Training involved 4 wk of cycling exercise inspiring either a normobaric normoxic (F(IO2) = approximately 20.9%) or a normobaric hypoxic (F(IO2) = approximately 16.0%) gas, respectively, in a double-blind manner. Cycling exercise was performed three times per week for 20-30 min at 70-85% of maximum heart rate determined either in normoxia or hypoxia. Resting plasma concentrations of blood lipids, lipoproteins, total homocysteine, and auscultatory arterial blood pressure responses at rest and in response to submaximal and maximal exercise were measured before and 4 d after physical training. Total power output during the training period was identical in both normoxic and hypoxic groups. Lean body mass increased by 1.4 +/- 1.5 kg following hypoxic training only (P < 0.001). While dietary composition and nutrient intake did not change during the study, both normoxic and hypoxic training decreased resting plasma concentrations of nonesterified fatty acids, total cholesterol, high density lipoprotein (HDL), and low density lipoprotein (LDL) (P < 0.05 - < 0.001). Apolipoproteins AI and B decreased following normoxic training only (P < or = 0.001). Plasma concentrations of resting total homocysteine decreased by 11% following hypoxic training (P < or = 0.05) and increased by 10% (P < 0.05) following normoxic training. These changes were independent of changes in serum vitamin B12 and red cell folate which remained stable throughout. A decreased lactate concentration during submaximal exercise was observed in response to both normoxic and hypoxic training. Hypoxic training decreased maximal systolic blood pressure by 10 +/- 9 mm Hg (P < 0.001) and the rate pressure product by 14 +/- 23 mm Hg x beats x min(-1)/100 (P < or = 0.001) and increased maximal oxygen uptake by 0.47 +/- 0.77 L x min(-1) (P < 0.05). Normoxic and hypoxic training was associated with significant improvements in selected risk factors and exercise capacity. The stimulus of intermittent normobaric hypoxia invoked an additive cardioprotective effect which may have important clinical implications.