Altered stress fibers and integrin expression in the Malpighian epithelium of Drosophila type IV collagen mutants.
Basement membranes (BMs) are highly specialized extracellular matrices (ECMs) that provide support and polarization cues for epithelial cells. Proper adhesion to the BM is pivotal in epithelial cell function and survival. Type IV collagens are the predominant components of all types of BMs, that form an irregular, polygonal lattice and serve as a scaffold for numerous other BM components and BM-associated cells. Mutations in the ubiquitous human BM components COL4A1 and COL4A2 cause a multisystem disorder involving nephropathy. Affected patients develop renal dysfunction and chronic kidney failure with or without hematuria. Mouse Col4a1 and Col4a2 mutants recapitulate the human symptoms. In vertebrates, excretion is accomplished by the kidneys and by the Malpighian tubules in insects, including the fruit fly Drosophila. Our present results with dominant, temperature-sensitive mutation of the Drosophila col4a1 gene demonstrate altered integrin expression and amplified effects of mechanical stress on the Malpighian epithelial cytoskeleton.


Simultaneous optical and electrical recording of single gramicidin channels.
We report here an approach for simultaneous fluorescence imaging and electrical recording of single ion channels in planar bilayer membranes. As a test case, fluorescently labeled (Cy3 and Cy5) gramicidin derivatives were imaged at the single-molecule level using far-field illumination and cooled CCD camera detection. Gramicidin monomers were observed to diffuse in the plane of the membrane with a diffusion coefficient of 3.3 x 10(-8) cm(2)s(-1). Simultaneous electrical recording detected gramicidin homodimer (Cy3/Cy3, Cy5/Cy5) and heterodimer (Cy3/Cy5) channels. Heterodimer formation was observed optically by the appearance of a fluorescence resonance energy transfer (FRET) signal (irradiation of Cy3, detection of Cy5). The number of FRET signals was significantly smaller than the number of Cy3 signals (Cy3 monomers plus Cy3 homodimers) as expected. The number of FRET signals increased with increasing channel activity. In numerous cases the appearance of a FRET signal was observed to correlate with a channel opening event detected electrically. The heterodimers also diffused in the plane of the membrane with a diffusion coefficient of 3.0 x 10(-8) cm(2)s(-1). These experiments demonstrate the feasibility of simultaneous optical and electrical detection of structural changes in single ion channels as well as suggesting strategies for improving the reliability of such measurements.


Integration of HIV testing services into family planning services: a systematic review.
Despite significant interest in integrating sexual and reproductive health (SRH) services into HIV services, less attention has been paid to linkages in the other direction. Where women and girls are at risk of HIV, offering HIV testing services (HTS) during their visits to family planning (FP) services offers important opportunities to address both HIV and unwanted pregnancy needs simultaneously. We conducted a systematic review of studies comparing FP services with integrated HTS to those without integrated HTS or with a lower level of integration (e.g., referral versus on-site services), on the following outcomes: uptake/counseling/offer of HTS, new cases of HIV identified, linkage to HIV care and treatment, dual method use, client satisfaction and service quality, and provider knowledge and attitudes about integrating HTS. We searched three online databases and included studies published in a peer-reviewed journal prior to the search date of June 20, 2017. Of 530 citations identified, six studies ultimately met the inclusion criteria. Three studies were conducted in Kenya, and one each in Uganda, Swaziland, and the USA. Most were in FP clinics. Three were from the Integra Initiative. Overall rigor was moderate, with one cluster-randomized trial. HTS uptake was generally higher with integrated sites versus comparison or pre-integration sites, including in adjusted analyses, though outcomes varied slightly across studies. One study found that women at integrated sites were more likely to have high satisfaction with services, but experienced longer waiting times. One study found a small increase in HIV seropositivity among female patients testing after full integration, compared to a dedicated HIV tester. No studies comparatively measured linkage to HIV care and treatment, dual method use, or provider knowledge/attitudes. Global progress and success for reaching SRH and HIV targets depends on progress in sub-Saharan Africa, where women bear a high burden of both unintended pregnancy and sexually transmitted infections, including HIV. While the evidence base is limited, it suggests that integration of HTS into FP services is feasible and has potential for positive joint outcomes. The success and scale-up of this approach will depend on population needs and health system factors.


2D and 3D Ultrasound in the evaluation of normal and abnormal fetal anatomy in the second and third trimesters in a level III center.
Between July 2000 and December 2003, a total number of 3,472 fetuses was evaluated by two-dimensional (2D) and three-dimensional (3D) ultrasonography. All examinations were carried out as part of a detailed level III ultrasound examination for fetal anomalies. The gestational age was between 11 and 35 weeks. A 3D endovaginal probe (5 - 7 MHz) was used for examinations between 11 and 13 weeks, and an abdominal 3D probe (5 MHz) after 13 weeks. Four different 3D image display modes were employed in visualizing fetal malformations: triplanar orthogonal display; surface display; transparent display; and the combined transparent and color display (= glass body-rendering). In 906 of the 3,472 high-risk pregnancies, fetuses with one to five fetal defects were found (mean 1.17). The total number of detected defects was 1,012, exclusive of 48 fetal heart defects. Fetal heart defects were excluded from this study since a reliable demonstration of these defects was not possible by 3D ultrasound. Comparing the 2D and 3D techniques, 3D sonography proved advantageous in 60.8 % of the defects, with the benefit derived from the exact tomographic survey using the multiplanar view in 69.9 % of these cases, from a more precise demonstration of the defect in the surface view in 25.2 %, from a distinct demonstration in the transparent view in 3.9 %, and from a precise demonstration in the combined transparent and color view in 1.0 %. In 42 of the 1,012 malformations (4.2 %), a defect was accurately identified or verified with 3D ultrasound only. 3D ultrasound proves not only a useful tool in appreciating the severity of a fetal defect, but also provides more convincing evidence of a normal fetus than conventional two-dimensional sonograms in cases with increased risk of a recurrent surface malformation.


Arginine, leucine, and glutamine stimulate proliferation of porcine trophectoderm cells through the MTOR-RPS6K-RPS6-EIF4EBP1 signal transduction pathway.
During the peri-implantation and early placentation periods in pigs, conceptuses (embryo and its extra-embryonic membranes) undergo dramatic morphological changes and differentiation that require the exchange of nutrients (histotroph) and gasses across the trophectoderm and a true epitheliochorial placenta. Of these nutrients, arginine (Arg), leucine (Leu), and glutamine (Gln) are essential components of histotroph; however, little is known about changes in their total amounts in the uterine lumen of cyclic and pregnant gilts and their effects on cell signaling cascades. Therefore, we determined quantities of Arg, Leu, and Gln in uterine luminal fluids and found that total recoverable amounts of these amino acids increased in pregnant but not cyclic gilts between Days 12 and 15 after onset of estrus. We hypothesized that Arg, Leu, and Gln have differential effects on hypertrophy, hyperplasia, and differentiated functions of trophectoderm cells that are critical to conceptus development. Primary porcine trophectoderm (pTr) cells treated with either Arg, Leu, or Gln had increased abundance of phosphorylated RPS6K, RPS6, and EIF4EBP1 compared to basal levels, and this effect was maintained for up to 120 min. When pTr cells were treated with Arg, Leu, and Gln, low levels of pRPS6K and pEIF4EBP1 were detected in the cytosol, but the abundance of nuclear pRPS6K increased. Immunofluorescence analyses revealed abundant amounts of pRPS6 protein in the cytoplasm of pTr cells treated with Arg, Leu, and Gln. These amino acids also increased proliferation of pTr cells. Furthermore, when Arg, Leu, and Gln were combined with siRNAs for either MTOR, RPTOR, or RICTOR, effects of those amino acids on proliferation of pTr cells were significantly inhibited. Collectively, these results indicate that Arg, Leu, and Gln act coordinately to stimulate proliferation of pTr cells through activation of the MTOR-RPS6K-RPS6-EIF4EBP1 signal transduction pathway.


Healthcare and medical graduates of 2009: their reactions to four key proposals in the Scottish Government's strategy for tackling alcohol misuse.
This study compares the views of final year medical, and nursing and allied health professional (NAHP) students in relation to four governmental proposals impacting on the sale and purchase of alcohol. Against a background of political will to address alcohol abuse in Scotland and moves within the National Health Service promoting a shifting of professional roles, self-completed questionnaires were administered in spring 2009 through course websites and lectures to final year medical and NAHP students. Questionnaires were returned by 406 NAHPs and 121 medical students. Over three quarters of all students agreed with the proposed change to reduce the drink driving limit to 50 mg/100 ml blood. Less support was evident for the raising of the minimum legal purchase age for off-sales (37%), the banning of below cost price promotions of alcohol (47%) and minimum retail pricing (37%). However, there were differences between the NAHP and medical students in the case of the final two proposals; over 60% of the medical students agreed they would have a positive impact. For NAHPs, figures were 41% and 31%, respectively. Support for four key proposals outlined by the Scottish Government to address alcohol misuse varied. Only the suggestion to lower the drink driving limit received backing overall and within students in these professions. Effectiveness of proposed restrictions on the price of alcohol was less well regarded except by medical students. Evident gaps in knowledge around health guidelines, and the finding that almost half of NAHPs disagreed that they had the appropriate knowledge to advise patients about responsible drinking advice and alcohol misuse problems, suggest a need for improved undergraduate education and continued professional development with respect to public health aspects of alcohol use.


Sudden infant death syndrome: seasonality and a biphasic model of pathogenesis.
This paper examines the relationship between season, age, and the sudden infant death syndrome (SIDS). It provides a theoretical model for the pathogenesis of SIDS and uses it as a framework to consider risk factor mechanism. A case series analysis was used to examine season and age in relation to SIDS and seasonal pattern and age at death distribution of perinatal risk factors. The source population for the SIDS cases in this study was all live births in the state of Tasmania, Australia, 1975 to 1987 inclusive. Cases were all infants born 1975 to 1987 who died of SIDS on whom birth notification information was available (n = 348). The live birth cohort 1980-87 (n = 55,944) was used as the control population for risk factor identification. The median ages of death for spring, summer, autumn, and winter born infants were 115, 103.5, 91 and 78 days. Spring and summer born infants died at a significantly older median age than winter born infants. The month of birth distribution of SIDS cases did not alter significantly from a uniform, nonseasonal distribution (p greater than 0.25) but month of death was seasonally distributed (p less than 0.01). Premature and low birthweight infants died at an older median age (p less than 0.05) than term and non-low-birthweight infants. An excess of male infant deaths and infant deaths to older mothers occurred during winter (p less than 0.05). The pathogenesis of SIDS can be represented as a biphasic model with three pathways of risk factor operation. In this study, season influenced the age at death of SIDS infants. We propose that risk factors with a strong seasonal distribution are likely to be operating in the postnatal period.


Abrogation of the Chk1-mediated G(2) checkpoint pathway potentiates temozolomide-induced toxicity in a p53-independent manner in human glioblastoma cells.
Temozolomide (TMZ) produces O(6)-methylguanine in DNA, which in turn mispairs with thymine, triggering futile DNA mismatch repair (MMR) and ultimately cell death. We found previously that in p53-proficient human glioma cells, TMZ-induced futile DNA MMR resulted not in apoptosis but rather in prolonged, p53- and p21-associated G(2)-M arrest and senescence. Additionally, p53-deficient cells were relatively more TMZ resistant than p53-deficient glioma cells, which underwent only transient G(2)-M arrest before death by mitotic catastrophe. These results suggested that prolonged G(2)-M arrest might protect cells from TMZ-induced cytotoxicity. In the present study, we therefore focused on the mechanism by which TMZ induces G(2)-M arrest and on whether inhibition of such G(2)-M arrest might sensitize glioma cells to TMZ-induced toxicity. U87MG glioma cells treated with TMZ underwent G(2)-M arrest associated with Chk1 activation and phosphorylation of both cdc25C and cdc2. These TMZ-induced effects were inhibited by the Chk1 kinase inhibitor UCN-01. Although not in itself toxic, UCN-01 increased the cytotoxicity of TMZ 5-fold, primarily by inhibiting cellular senescence and increasing the percentage of cells bypassing G(2)-M arrest and undergoing mitotic catastrophe. In addition to enhancing TMZ-induced cytotoxicity in p53-proficient cells, UCN-01 also blocked TMZ-induced Chk1 activation and transient G(2)-M arrest in p53-deficient U87MG-E6 cells and similarly enhanced TMZ-induced mitotic catastrophe and cell death. Taken together, these results indicate that Chk1 links TMZ-induced MMR to G(2)-M arrest. Furthermore, inhibition of the cytoprotective G(2) arrest pathway sensitizes cells to TMZ-induced cytotoxicity and may represent a novel, mechanism-based means of increasing TMZ efficacy in both p53 wild-type and p53 mutant glioma cells.


MUC4, a novel immunohistochemical marker identified by gene expression profiling, differentiates pleural sarcomatoid mesothelioma from lung sarcomatoid carcinoma.
Sarcomatoid mesothelioma, a histological subtype of malignant pleural mesothelioma, is a very aggressive tumor with a poor prognosis. Histological diagnosis of sarcomatoid mesothelioma largely depends on the histomorphological feature of spindled tumor cells with immunohistochemical reactivity to cytokeratins. Diagnosis also requires clinico-radiological and/or macroscopic evidence of an extrapulmonary location to differentiate it from lung sarcomatoid carcinoma. Although there are promising immunohistochemical antibody panels to differentiate mesothelioma from lung carcinoma, a consensus on the immunohistochemical markers that distinguish sarcomatoid mesothelioma from lung sarcomatoid carcinoma has not been reached and requires further study. We performed whole gene expression analysis of formalin-fixed paraffin-embedded tissue from sarcomatoid mesothelioma and lung sarcomatoid carcinoma and observed significant differences in the expression of MUC4 and other genes between sarcomatoid mesothelioma and lung sarcomatoid carcinoma. Immunohistochemistry demonstrated that MUC4 was expressed in the spindled tumor cells of lung sarcomatoid carcinoma (21/29, 72%) but was not expressed in any sarcomatoid mesothelioma (0/31, 0%). To differentiate sarcomatoid mesothelioma from lung sarcomatoid carcinoma, negative MUC4 expression showed 100% sensitivity and 72% specificity and accuracy rate of 87%, which is higher than immunohistochemical markers such as calretinin, D2-40 and Claudin-4. Therefore, we recommend to include MUC4 as a novel and useful negative immunohistochemical marker for differentiating sarcomatoid mesothelioma from lung sarcomatoid carcinoma.


Long-chain polyunsaturated fatty acids and the pathophysiology of myalgic encephalomyelitis (chronic fatigue syndrome).
Evidence is put forward to suggest that myalgic encephalomyelitis, also known as chronic fatigue syndrome, may be associated with persistent viral infection. In turn, such infections are likely to impair the ability of the body to biosynthesise n-3 and n-6 long-chain polyunsaturated fatty acids by inhibiting the delta-6 desaturation of the precursor essential fatty acids--namely, alpha-linolenic acid and linoleic acid. This would, in turn, impair the proper functioning of cell membranes, including cell signalling, and have an adverse effect on the biosynthesis of eicosanoids from the long-chain polyunsaturated fatty acids dihomo-gamma-linolenic acid, arachidonic acid and eicosapentaenoic acid. These actions might offer an explanation for some of the symptoms and signs of myalgic encephalomyelitis. A potential therapeutic avenue could be offered by bypassing the inhibition of the enzyme delta-6-desaturase by treatment with virgin cold-pressed non-raffinated evening primrose oil, which would supply gamma-linolenic acid and lipophilic pentacyclic triterpenes, and with eicosapentaenoic acid. The gamma-linolenic acid can readily be converted into dihomo-gamma-linolenic acid and thence arachidonic acid, while triterpenes have important free radical scavenging, cyclo-oxygenase and neutrophil elastase inhibitory activities. Furthermore, both arachidonic acid and eicosapentaenoic acid are, at relatively low concentrations, directly virucidal.


A SEER-Medicare population-based study of lymphedema-related claims incidence following breast cancer in men.
Each year there are an estimated 200,000 new breast cancer cases diagnosed in the United States; of these, 1% of cases are in men. Lymphedema can be a devastating complication from breast cancer and its treatment. Currently, almost all lymphedema-related research is based on women and extrapolated to men. We conducted the first population-based study of men with incident breast cancer of any stage, diagnosed from 1998 to 2005, who were 65 years and older in the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database. We utilized claims related to lymphedema and lymphedema treatment in our cohort. We defined 'lymphedema-specific' claims to contain the word 'lymphedema' in the Medicare claim description; similarly, 'lymphedema-related' claims were defined as treatments reimbursed for lymphedema but not necessarily containing the word 'lymphedema' in the Medicare claim description. We identified 628 men with incident breast cancer from 1998 to 2005 who were 65 years and older. The cumulative incidence, censored for deaths, of lymphedema-specific claims at 2, 3, 4, and 5 years was 8.0, 9.2, 10.5, and 10.5%, respectively. The median follow-up was 3.4 years and for those without any event was 4.7 years. The cumulative incidence, censored for deaths, of lymphedema-related claims at 2, 3, 4, and 5 years was 26.9, 32.2, 35.4, and 39.8%. Rates for men were similar to analogous rates for women. Lymphedema is a common complication affecting men with breast cancer as well as women and appropriate treatment and rehabilitation strategies need to be implemented for both genders.


Characterization of the CCR2 chemokine receptor: functional CCR2 receptor expression in B cells.
We have derived anti-human CCR2-specific mAbs by immunization with synthetic peptides corresponding to CCR2 sequences presumably involved in the interaction with its ligand(s). The characterization of these mAbs includes the ability to recognize the CCR2 receptor specifically, as well as the function based on their ability to promote Ca2+ influx or to block MCP-1-induced Ca2+ influx and chemotaxis. One mAb (MCP-1 R02) that is directed to the NH2 terminal domain of the CCR2 receptor has MCP-1 agonist activity, and two that recognize the third extracellular domain (MCP-1R04 and MCP-1 R05) have MCP-1 antagonist activity. We analyzed the presence of CCR2 in several PBL and tonsil-derived leukocyte populations and found expression of this receptor in monocytes, activated T cells, and, surprisingly, in B cells. CCR2 receptor expression in B cells was further corroborated in Southern blot using CCR2-specific probes. Moreover, both MCP-1 and the agonist mAb trigger specific B cell migration via a PTX-sensitive mechanism, indicating the presence of a functional CCR2 receptor in these cells.


Impact of a pertussis epidemic on infant vaccination in Washington state.
Washington State experienced a pertussis epidemic from October 2011 to December 2012. There was wide variation in incidence by county. The objectives of this study were to determine how the pertussis epidemic affected infant vaccination in Washington State and whether the incidence in counties modified this effect. We conducted an ecologic before-after study to compare the proportion of infants up to date (UTD) with a pertussis-containing vaccine at time points before (September 30, 2011), during (September 30, 2012), and after (September 30, 2013) the epidemic. Children aged 3 to 8 months enrolled in the Washington State Immunization Information System with documented county of residence were included. UTD status was determined as ≥ 1, ≥ 2, or ≥ 3 doses of a pertussis-containing vaccine at ages 3, 5, and 7 months, respectively. Generalized linear models with extension to the binomial family and clustered robust standard errors were used to examine differences in the proportion of UTD infants between preepidemic and either epidemic or postepidemic points. The potential modifying effect of pertussis incidence by county was examined. We found no significant difference in statewide UTD status with a pertussis-containing vaccine between preepidemic and either epidemic (absolute difference 2.1%; 95% confidence interval, -1.6 to 5.9) or postepidemic (absolute difference 0.2%; 95% confidence interval, -4.0 to 4.5) time points. There was no significant modification by county pertussis incidence. There was wide variation in the absolute difference in UTD status across counties. A statewide pertussis epidemic does not appear to have significantly changed the proportion of infants who were UTD with a pertussis-containing vaccine.


Lethal toxicity of lipopolysaccharide and tumor necrosis factor in normal and D-galactosamine-treated mice.
The toxic properties of human recombinant tumor necrosis factor (TNF) were investigated in mice made hypersensitive to endotoxin by treatment with D-galactosamine. C3H/TifF mice treated with D-galactosamine were rendered sensitive to the lethal effects of submicrogram amounts of TNF. In the absence of D-galactosamine, TNF caused approximately 80% lethality with 500 micrograms. The duration of sensitization to TNF lasted up to 8 h after D-galactosamine administration, that towards LPS, up to 4 h. As with LPS, with TNF sensitization could be inhibited by uridine administered up to 2 h after D-galactosamine/TNF, showing that the early biochemical alterations in the liver known to be necessary for sensitization to LPS are also necessary for sensitization to TNF. In contrast to LPS, the toxicity of TNF was expressed also in D-galactosamine-treated endotoxin-resistant C3H/HeJ mice. The susceptibility of these mice to TNF was identical to that of endotoxin sensitive mice. In the absence of D-galactosamine the toxicity of TNF in C3H/HeJ mice was comparable to that obtained in C3H/TifF mice, being lethal with amounts of the order of 500 micrograms. The present results support the hypothesis that TNF is a mediator of lethal toxicity of endotoxin.


Single-molecule sequencing resolves the detailed structure of complex satellite DNA loci in Drosophila melanogaster.
Highly repetitive satellite DNA (satDNA) repeats are found in most eukaryotic genomes. SatDNAs are rapidly evolving and have roles in genome stability and chromosome segregation. Their repetitive nature poses a challenge for genome assembly and makes progress on the detailed study of satDNA structure difficult. Here, we use single-molecule sequencing long reads from Pacific Biosciences (PacBio) to determine the detailed structure of all major autosomal complex satDNA loci in Drosophila melanogaster, with a particular focus on the 260-bp and Responder satellites. We determine the optimal de novo assembly methods and parameter combinations required to produce a high-quality assembly of these previously unassembled satDNA loci and validate this assembly using molecular and computational approaches. We determined that the computationally intensive PBcR-BLASR assembly pipeline yielded better assemblies than the faster and more efficient pipelines based on the MHAP hashing algorithm, and it is essential to validate assemblies of repetitive loci. The assemblies reveal that satDNA repeats are organized into large arrays interrupted by transposable elements. The repeats in the center of the array tend to be homogenized in sequence, suggesting that gene conversion and unequal crossovers lead to repeat homogenization through concerted evolution, although the degree of unequal crossing over may differ among complex satellite loci. We find evidence for higher-order structure within satDNA arrays that suggest recent structural rearrangements. These assemblies provide a platform for the evolutionary and functional genomics of satDNAs in pericentric heterochromatin.


Neutrophilic airways inflammation in lung cancer: the role of exhaled LTB-4 and IL-8.
Recent advances in lung cancer biology presuppose its inflammatory origin. In this regard, LTB-4 and IL-8 are recognized to play a crucial role in neutrophil recruitment into airways during lung cancer.Notwithstanding the intriguing hypothesis, the exact role of neutrophilic inflammation in tumour biology remains complex and not completely known.The aim of this study was to give our contribution in this field by investigating LTB-4 and IL-8 in the breath condensate of NSCLC patients and verifying their role in cancer development and progression. We enrolled 50 NSCLC patients and 35 controls. LTB-4 and IL-8 concentrations were measured in the breath condensate and the blood of all the subjects under study using EIA kits. Thirty NSCLC patients and ten controls underwent induced sputum collection and analysis. LTB-4 and IL-8 resulted higher in breath condensate and the blood of NSCLC patients compared to controls. Significantly higher concentrations were found as the cancer stages progressed. A positive correlation was observed between exhaled IL-8 and LTB-4 and the percentage of neutrophils in the induced sputum. The high concentrations of exhaled LTB-4 and IL-8 showed the presence of a neutrophilic inflammation in the airways of NSCLC patients and gave a further support to the inflammatory signalling in lung cancer. These exhaled proteins could represent a suitable non-invasive marker in the diagnosis and monitoring of lung cancer.


Viral burden and disease progression in rhesus monkeys infected with chimeric simian-human immunodeficiency viruses.
To determine the role of viral burden in simian-human immunodeficiency virus (SHIV)-induced disease, cellular provirus and plasma viral RNA levels were measured after inoculation of rhesus monkeys with four different SHIVs. These SHIVs included SHIV-HXBc2 and SHIV-89.6, constructed with env, tat, rev, and vpu derived from either cell line-passaged or primary patient isolates of human immunodeficiency virus type 1; the viral quasispecies SHIV-89.6P derived after in vivo passage of SHIV-89.6; and a molecular clone, SHIV-KB9, derived from SHIV-89.6P. SHIV-HXBc2 and SHIV-89.6 are nonpathogenic in rhesus monkeys; SHIV-89.6P and SHIV-KB9 cause rapid CD4(+) T cell depletion and an immunodeficiency syndrome. Relative SHIV provirus levels were highest during primary infection in monkeys infected with SHIV-89.6P, the virus that caused the most rapid and dramatic CD4(+) T cell depletion. However, by 10 weeks postinoculation, provirus levels were similar in monkeys infected with the pathogenic and nonpathogenic chimeric viruses. The virus infections that resulted in the highest peak and chronic viral RNA levels were the pathogenic viruses SHIV-89.6P and SHIV-KB9. SHIV-89. 6P uniformly caused rapid and profound CD4(+) T cell depletion and immunodeficiency. Infection with the SHIV-KB9 resulted in very low CD4(+) T cell counts without seroconversion in some monkeys and a substantial but less profound CD4(+) T cell depletion and rapid seroconversion in others. Surprisingly, the level of plasma viremia did not differ between SHIV-KB9-infected animals exhibiting these contrasting outcomes, suggesting that host factors may play an important role in AIDS virus pathogenesis.


Enhanced photo-responsiveness in a photoswitchable system model: emergent hormetic catalysis.
Michaelis Menten catalysis by a T-photochromic system has been analyzed numerically. Using an appropriate set of rate constants and quantum yields, we have evidenced an enhanced photo-responsiveness at a medium light intensity: the plot of the initial rate vs. light intensity is bell-shaped. This emergent phenomenon can be qualified as hormetic catalysis. The analysis of the chemical flows shows that a directional rotation occurs within the cyclic scheme. Non equilibrium conditions are provided by two independent sources: the chemical energy dissipation from the irreversible exergonic reaction and the steady transformation of light into heat by T-photochromism. A literature survey, showing that most of the required kinetic features are not so rare, let us anticipate its practical feasibility.


Phase I pharmacokinetic study of tea polyphenols following single-dose administration of epigallocatechin gallate and polyphenon E.
Green tea has been shown to exhibit cancer-preventive activities in preclinical studies. Its principal active components include epigallocatechin gallate (EGCG), epigallocatechin (EGC), epicatechin (EC), and epicatechin gallate, of which EGCG is the most abundant and possesses the most potent antioxidative activity. We performed a Phase I pharmacokinetic study to determine the systemic availability of green tea catechins after single oral dose administration of EGCG and Polyphenon E (decaffeinated green tea catechin mixture). Twenty healthy subjects (five subjects/dose level) were randomly assigned to one of the dose levels (200, 400, 600, and 800 mg based on EGCG content). All subjects were randomly crossed-over to receive the two catechin formulations at the same dose level. Blood and urine samples were collected for up to 24 h after oral administration of the study medication. Tea catechin concentrations in plasma and urine samples were determined using high-performance liquid chromatography with the coulometric electrode array detection system. After EGCG versus Polyphenon E administration, the mean area under the plasma concentration-time curves (AUC) of unchanged EGCG were 22.5 versus 21.9, 35.4 versus 52.2, 101.9 versus 79.7, and 167.1 versus 161.4 min x microg/ml at the 200-, 400-, 600-, and 800-mg dose levels, respectively. EGC and EC were not detected in plasma after EGCG administration and were present at low/undetectable levels after Polyphenon E administration. High concentrations of EGC and EC glucuronide/sulfate conjugates were found in plasma and urine samples after Polyphenon E administration. There were no significant differences in the pharmacokinetic characteristics of EGCG between the two study medications. The AUC and maximum plasma concentration (Cmax) of EGCG after the 800-mg dose of EGCG were found to be significantly higher than those after the 200- and 400-mg dose. The AUC and Cmax of EGCG after the 800-mg dose of Polyphenon E were significantly higher than those after the three lower doses. We conclude that the two catechin formulations resulted in similar plasma EGCG levels. EGC and EC were present in the body after the Polyphenon E administration; however, they were present predominantly in conjugated forms. The systemic availability of EGCG increased at higher doses, possibly due to saturable presystemic elimination of orally administered green tea polyphenols.


Active and legacy mining in an arid urban environment: challenges and perspectives for Copiapó, Northern Chile.
Urban expansion in areas of active and legacy mining imposes a sustainability challenge, especially in arid environments where cities compete for resources with agriculture and industry. The city of Copiapó, with 150,000 inhabitants in the Atacama Desert, reflects this challenge. More than 30 abandoned tailings from legacy mining are scattered throughout its urban and peri-urban area, which include an active copper smelter. Despite the public concern generated by the mining-related pollution, no geochemical information is currently available for Copiapó, particularly for metal concentration in environmental solid phases. A geochemical screening of soils (n = 42), street dusts (n = 71) and tailings (n = 68) was conducted in November 2014 and April 2015. Organic matter, pH and elemental composition measurements were taken. Notably, copper in soils (60-2120 mg/kg) and street dusts (110-10,200 mg/kg) consistently exceeded international guidelines for residential and industrial use, while a lower proportion of samples exceeded international guidelines for arsenic, zinc and lead. Metal enrichment occurred in residential, industrial and agricultural areas near tailings and the copper smelter. This first screening of metal contamination sets the basis for future risk assessments toward defining knowledge-based policies and urban planning. Challenges include developing: (1) adequate intervention guideline values; (2) appropriate geochemical background levels for key metals; (3) urban planning that considers contaminated areas; (4) cost-effective control strategies for abandoned tailings in water-scarce areas; and (5) scenarios and technologies for tailings reprocessing. Assessing urban geochemical risks is a critical endeavor for areas where extreme events triggered by climate change are likely, as the mud flooding that impacted Copiapó in late March 2015.


Ischemia-reperfusion injury and cardioprotection: investigating PTEN, the phosphatase that negatively regulates PI3K, using a congenital model of PTEN haploinsufficiency.
Activation of the PI3K/Akt pathway protects the heart from ischemia-reperfusion injury (IRI). The phosphatase PTEN is the main negative regulator of this pathway. We hypothesized that reduced PTEN levels could protect against IRI. Isolated perfused mouse hearts from PTEN(+/-) and their littermates PTEN(+/+) (WT), were subjected to 35 min global ischemia and 30 min reperfusion, with and without 2, 4 or 6 cycles ischemic preconditioning (IPC). The end point was infarct size, expressed as a percentage of the myocardium at risk (I/R%). PTEN and Akt levels were determined using Western blot analysis. Unexpectedly, there were no significant differences in infarction between PTEN(+/-) and WT (42.1 +/- 5.0% Vs. 45.6 +/- 3.3%). However, the preconditioning threshold was significantly reduced in the PTEN(+/-) Vs. WT, with 4 cycles of IPC being sufficient to reduce I/R%, compared to 6 cycles in the WT (4 cycles IPC: 29.8. +/- 3.69% in PTEN(+/-) Vs. 45.5. +/- 5.08% in WT, P < 0.01). In addition, the ratio between the phospho/total Akt (Ser473 and Thr308) was slightly but significantly increased in the PTEN(+/-) indicating an upregulation of PI3K/Akt pathway. Interestingly, the levels of the other phosphatases that may negatively regulate the PI3K/Akt pathway (PP2A, SHIP2 and PHLPP) were not significantly different between littermates and PTEN(+/-). In conclusion, PTEN haploinsufficiency alone does not induce cardioprotection in this model; however, it reduces the threshold of protection induced by IPC.


Histologically proven, low-grade brainstem gliomas in children: 30-year experience with long-term follow-up at Mayo Clinic.
To evaluate long-term overall survival (OS), progression-free survival (PFS), and outcomes in pathologically proven brainstem low-grade gliomas (BS-LGG) in children. The Mayo Clinic tumor registry identified 48 consecutive children (≤20 y, 52% female) with biopsy-proven BS-LGG treated at Mayo Clinic between January 1971 and December 2004. Medical records were retrospectively reviewed. For analysis, patients were censored at the time of recurrence, death, or last follow-up. The median age at diagnosis was 12 years with a median follow-up of 6.0 years. The majority of tumors were grade I (69%) and pathology was consistent with an astrocytoma in the majority of patients (98%). Gross total resection was obtained in 4, subtotal in 17, and 27 patients were biopsied only. Postoperative radiotherapy (RT) was used in 29 patients. Median OS for the entire group was 14.8 years with a 1-, 5-, and 10-year OS of 85%, 67% and 59%, respectively. Median PFS for the entire group was 7.3 years. Improved survival was associated with undergoing resection versus biopsy-only with 5-year OS rates of 85% and 50% (P=0.002), respectively. A high proportion of patients (42%) had diffuse tumors and 13 patients (27%) had diffuse pontine gliomas (DPGs). DPGs had an OS of 1.8 years with a worse median PFS than non-DPGs (1.8 vs. 11.1 y; P=0.009). RT was used preferentially in patients with poor prognosis such as those who had a biopsy-only procedure (19/27) and DPGs (9/13). OS in this single institution retrospective study in pathologically proven BS-LGG with extensive follow-up displayed favorable long-term outcomes. Improved outcomes were associated with nondiffuse classification.


Green Communication for Wireless Body Area Networks: Energy Aware Link Efficient Routing Approach.
Recent technological advancement in wireless communication has led to the invention of wireless body area networks (WBANs), a cutting-edge technology in healthcare applications. WBANs interconnect with intelligent and miniaturized biomedical sensor nodes placed on human body to an unattended monitoring of physiological parameters of the patient. These sensors are equipped with limited resources in terms of computation, storage, and battery power. The data communication in WBANs is a resource hungry process, especially in terms of energy. One of the most significant challenges in this network is to design energy efficient next-hop node selection framework. Therefore, this paper presents a green communication framework focusing on an energy aware link efficient routing approach for WBANs (ELR-W). Firstly, a link efficiency-oriented network model is presented considering beaconing information and network initialization process. Secondly, a path cost calculation model is derived focusing on energy aware link efficiency. A complete operational framework ELR-W is developed considering energy aware next-hop link selection by utilizing the network and path cost model. The comparative performance evaluation attests the energy-oriented benefit of the proposed framework as compared to the state-of-the-art techniques. It reveals a significant enhancement in body area networking in terms of various energy-oriented metrics under medical environments.


Rapid and sensitive detection of neuron specific enolase with a polydopamine coated plasmonic chip utilizing a rear-side coupling method.
A rapid and sensitive detection of a cancer marker, neuron specific enolase (NSE), is demonstrated by using a disposable silver plasmonic chip functionalized with a mussel-inspired polydopamine (PDA) coating. A plasmonic chip consisting of a diffraction grating coated with a silver thin film is used for the excitation of propagating surface plasmon resonance through a rear-side grating coupling method. Simple and quick bio-functionalization of the sensor surface is performed by PDA coating which requires 20 min for deposition, and allows direct attachment of the capture antibody without using any coupling agents. A fluorescence based sandwich immunoassay is used for the detection of NSE by utilizing surface plasmon enhanced fluorescence (SPF) spectroscopy. The developed biosensor scheme provides approximately linear sensor responses for the sample containing NSE with the concentration around the clinically important value (12 ng mL-1) in both buffer and diluted human serum (25 vol% to a buffer solution). The detection limit for NSE is 0.5 ng mL-1 (11 pM) and 1.4 ng mL-1 (30 pM) in a buffer solution and diluted human serum, respectively. The presented biosensor scheme requires a small amount of the sample down to 10 μL in human serum and a short incubation time (15 min) of the sample solution containing NSE, enabling less invasive and rapid detection of NSE. This is the first example of the sensitive sandwich immunoassay demonstrated by using a plasmonic chip for the measurement of the sample dissolved in a complex medium with a rear side coupling method, which progresses the universal use of the SPF biosensors with a disposable plasmonic chip.


Characteristics and prognostic value of papillary histologic subtype in nonmetastatic renal cell carcinoma in Korea: a multicenter study.
To analyze the characteristics of nonmetastatic papillary renal cell carcinomas (RCC) and the prognostic value of RCC histologic subtyping, based on a large multicenter experience in Korea. A total of 2,905 patients with nonmetastatic RCC (TxN0M0) at the time of surgery were retrospectively enrolled from five institutions between 1999 and 2011 in Korea. Among these, patients with clear cell subtype (n = 2,488, 85.6%) and papillary subtype (n = 192, 6.6%) were included in our study. Patients with papillary subtype did not differ significantly from those with clear cell subtype on the following parameters: age (P = .694), gender (P = .511), body mass index (P = .136), patient performance status (P = .419), symptoms at presentation (P = .419), tumor size (P = .778) and pathologic stage (P = .367). However, high Fuhrman's grades were more common in papillary subtypes compared with clear cell subtypes (P = .001). The 5-year recurrence-free survival rates in patients with clear cell subtype and papillary subtype were 84.9% and 86.7%, respectively (P = .167). The 5-year cancer-specific survival rates in patients with clear cell subtype and papillary subtype were 92.0% and 93.1%, respectively (P = .931). Histologic subtype was not an independent prognostic factor of recurrence-free and cancer-specific survival (P = .107 and P = .998, respectively). Our study suggests that the characteristics and prognosis of papillary subtype might be comparable to those of clear cell subtype in non-metastatic renal cell carcinoma, especially in Asia.


Selective roles of normal and mutant huntingtin in neural induction and early neurogenesis.
Huntington's disease (HD) is a neurodegenerative disorder caused by abnormal polyglutamine expansion in the amino-terminal end of the huntingtin protein (Htt) and characterized by progressive striatal and cortical pathology. Previous reports have shown that Htt is essential for embryogenesis, and a recent study by our group revealed that the pathogenic form of Htt (mHtt) causes impairments in multiple stages of striatal development. In this study, we have examined whether HD-associated striatal developmental deficits are reflective of earlier maturational alterations occurring at the time of neurulation by assessing differential roles of Htt and mHtt during neural induction and early neurogenesis using an in vitro mouse embryonic stem cell (ESC) clonal assay system. We demonstrated that the loss of Htt in ESCs (KO ESCs) severely disrupts the specification of primitive and definitive neural stem cells (pNSCs, dNSCs, respectively) during the process of neural induction. In addition, clonally derived KO pNSCs and dNSCs displayed impaired proliferative potential, enhanced cell death and altered multi-lineage potential. Conversely, as observed in HD knock-in ESCs (Q111 ESCs), mHtt enhanced the number and size of pNSC clones, which exhibited enhanced proliferative potential and precocious neuronal differentiation. The transition from Q111 pNSCs to fibroblast growth factor 2 (FGF2)-responsive dNSCs was marked by potentiation in the number of dNSCs and altered proliferative potential. The multi-lineage potential of Q111 dNSCs was also enhanced with precocious neurogenesis and oligodendrocyte progenitor elaboration. The generation of Q111 epidermal growth factor (EGF)-responsive dNSCs was also compromised, whereas their multi-lineage potential was unaltered. These abnormalities in neural induction were associated with differential alterations in the expression profiles of Notch, Hes1 and Hes5. These cumulative observations indicate that Htt is required for multiple stages of neural induction, whereas mHtt enhances this process and promotes precocious neurogenesis and oligodendrocyte progenitor cell elaboration.


Impact of bottle size on in-home consumption of wine: feasibility and acceptability randomised cross-over study.
Reducing alcohol consumption across populations would prevent many non-communicable diseases. Large packages increase food and non-alcoholic drink consumption and large glasses increase wine consumption. Smaller bottles may reduce alcohol consumption but their impact is uncertain. This study aims to (i) explore the feasibility and acceptability of conducting a large-scale randomised study to assess the impact of bottle size on in-home wine consumption and (ii) estimate the effect size and variance of the intervention on consumption to inform the design of future studies. Cross-over randomised study in which 16 households in Cambridge, England, consuming at least two 750-ml bottles of wine each week, received a pre-set volume of wine biweekly for 4 weeks, in 750-ml and 375-ml bottles, in random order. Consumption was assessed by recording the number of empty and partially full bottles at the end of each biweekly period. At the end of the study, household representatives were interviewed about their experiences of participating in the study. The study procedures proved feasible. Comparable to similar trials, 14% of identified eligible households (18/125) consented to participate in the study. Attrition between consent and study completion was 11% (2/18) and 0% between study periods and 13% of households (2/16) correctly identified the study aim. The study procedures were considered acceptable. After adjusting for guest and out-of-home consumption, the difference in consumption between the 750-ml (3385.2 ml; SD = 1698.5) and 375-ml bottles (3376.7 ml; SD = 1719.0) was 8.4 ml (SD = 1235.4; 95%CI - 596.9, 613.8). Results suggest a possible order effect, with households receiving the 375-ml bottles first consuming more wine out of the 750-ml bottles and vice versa. This might also reflect an increase in consumption with study duration. Households receiving the 375-ml bottles first (6315.9 ml; SD = 3293.5) also drank less wine overall than those receiving the 750-ml bottles first (7335.4 ml; SD = 3735.4). The findings support the feasibility and acceptability of running a large-scale randomised study to assess the impact of bottle size on in-home wine consumption. Due to the heterogeneous patterning of results, a future study will be powered using the variance observed in the current study to detect a meaningful reduction of 250 ml of wine when consumed from smaller compared with larger bottles. Open Science Framework (OSF): rmk43; May 23, 2017.


Epilepsy-associated stigma in sub-Saharan Africa: the social landscape of a disease.
Many studies in developed regions of the world have confirmed that stigma contributes substantially to the psychological and social burden of epilepsy. Relatively few studies of epilepsy-associated stigma have been conducted in Africa, where much of the world's burden of epilepsy exists. In sub-Saharan Africa (SSA), particularly in rural regions, close family ties, communal living situations, and traditional belief systems undoubtedly influence the expression of stigmatization. A review of the epidemiologic, anthropologic, and sociologic studies of epilepsy in SSA provides significant insights into how people with epilepsy (PWE) are perceived by their communities and families and how these perceptions translate into limited social and economic opportunities and possibly worsen the physical vulnerability of PWE in this region. The medical community is not exempt from the social process of stigmatization, and poor public health infrastructure and medical services undoubtedly contribute to the cycle of epilepsy-associated stigma through wide treatment gaps, poor seizure control, and high rates of seizure-related injury. In this review, we extrapolate data from existing studies of epilepsy in SSA coupled with our own experience providing epilepsy care in the region to give an overview of the social landscape of this common, devastating condition.


Pattern and clinical characteristics of firearm injuries.
To determine the pattern and characteristics of patients admitted with firearm injuries (FAIs) and establish the morbidity and mortality associated with these injuries. Retrospective cross-sectional study. Kenyatta National Hospital (KNH), January 2004 to December 2005. All patients admitted with physically evident firearm injury. There were a total of 717 patients recorded with FAIs constituting 0.6% of the total number of patients seen in the casualty. Of these, 421 (58.7%) were admitted and treated as in- patients. A firearm was used in 6.7% of the 6300 assault cases recorded in 2004 and in 9.7% of the 3079 cases recorded in 2005. The increase from 6.7% in 2004 to 9.7% in 2005 was statistically significant (p < 0.05). There were 370 (87.9%) males and 49 (11.6%) females giving a male to female ratio of 7.5:1. The mean age was 29.7 +/- 10.9 years with a range of 3 to 66 years. At least 262 (62.2%) of the 421 admitted FAI casualties were treated under general anaesthesia (GA). The average duration of operation per patient was 2 +/- 1.5 hours. FAIs are on the increase and affect all age groups but is largely a disease of a young male adult in the 3rd and 4th decade of life. Mortality is higher with increasing age while female victims are fewer but on average six years younger than males. The lower extremities are the commonest target among the survivors. However, abdominal wounds tend to be the most lethal, accounting for greater mortality.


The feasibility of parameterizing four-state equilibria using relaxation dispersion measurements.
Coupled equilibria play important roles in controlling information flow in biochemical systems, including allosteric molecules and multidomain proteins. In the simplest case, two equilibria are coupled to produce four interconverting states. In this study, we assessed the feasibility of determining the degree of coupling between two equilibria in a four-state system via relaxation dispersion measurements. A major bottleneck in this effort is the lack of efficient approaches to data analysis. To this end, we designed a strategy to efficiently evaluate the smoothness of the target function surface (TFS). Using this approach, we found that the TFS is very rough when fitting benchmark CPMG data to all adjustable variables of the four-state equilibria. After constraining a portion of the adjustable variables, which can often be achieved through independent biochemical manipulation of the system, the smoothness of TFS improves dramatically, although it is still insufficient to pinpoint the solution. The four-state equilibria can be finally solved with further incorporation of independent chemical shift information that is readily available. We also used Monte Carlo simulations to evaluate how well each adjustable parameter can be determined in a large kinetic and thermodynamic parameter space and how much improvement can be achieved in defining the parameters through additional measurements. The results show that in favorable conditions the combination of relaxation dispersion and biochemical manipulation allow the four-state equilibrium to be parameterized, and thus coupling strength between two processes to be determined.


Measurement of red blood cell aggregation in disposable capillary tubes.
A new method is described in this paper that allows measurement of red blood cell (RBC) aggregation indexes in disposable glass tubes within minutes. Light transmission through the RBC suspension filled into a microhematocrit capillary at stasis is recorded during RBC aggregation; a novel method assures an initial dispersion of aggregates in the capillary. The resulting light transmittance-time data are analyzed to calculate various parameters. Measurement of erythrocyte sedimentation rate (ESR) and RBC aggregation using well established methods and the newly developed capillary tube aggregometer in blood samples with a wide range of RBC aggregation indicated significant correlations between these parameters. Additionally, light transmittance during complete disaggregation allows estimating hematocrit, thereby enabling hematocrit correction of the measured and calculated parameters. The newly developed capillary tube RBC aggregometer is suitable for use as a method to rapidly monitor disease activity and the acute phase response, especially at the point-of-care (e.g., health care facilities, physician's office) and for field studies.


Domain-general and domain-specific influences on emerging numerical cognition: Contrasting uni-and bidirectional prediction models.
Domain-general skills such as executive functions (EFs), and domain-specific skills such as non-symbolic number sense and symbolic understanding are often pitted against each other as predictors of emerging maths. Here we aimed to investigate early childhood relations between these foundational skills with a balanced, longitudinal design. One hundred and seventy 3- and 4-year-old-children were tested at two time points, 5 months apart, on four domain-general executive and five domain-specific numeracy tasks. A latent EF factor was a strong predictor of symbolic maths and of their growth. In addition, stronger symbolic maths at Time 1 was correlated with later stronger EF, but symbolic maths did not predict EF growth. Our findings provide novel insights into dynamic interplay between general and specific cognitive skills contributing to preschool maths.


Hemoglobin Hakkari: an autosomal dominant form of beta thalassemia with inclusion bodies arising from de novo mutation in exon 2 of beta globin gene.
Certain beta globin gene mutations produce a thalassemia major phenotype in the heterozygous state. While most such patients have thalassemia intermedia, we describe a young Guatemalan child with a de novo mutation in the beta globin gene, codon 31 T --> G (Hemoglobin Hakkari), who developed severe anemia at the age of 10 months and remains transfusion-dependent. The substitution of B13 leucine with arginine in the beta globin results in alteration of a critical heme contact point resulting in an extremely unstable variant hemoglobin and a clinical picture that is characterized by ineffective erythropoiesis and numerous intracytoplasmic inclusions within the erythrocyte precursors of the bone marrow. .


Trophic Transfer and Transformation of CeO2 Nanoparticles along a Terrestrial Food Chain: Influence of Exposure Routes.
The trophic transfer and transformation of CeO2 nanoparticles (NPs) through a simulated terrestrial food chain were investigated using a radiotracer technique and X-ray absorption near edge structure (XANES). Radioactive 141CeO2 NPs were applied to head lettuce ( Lactuca sativa), treated via root exposure in its potting soil (5.5 or 11 mg/plant) for 30 days or foliar exposure (7.2 mg/plant, with half of the leaves treated and the other half not) for 7 days. Subsequently, two groups of land snails ( Achatina fulica) were exposed to 141Ce via either a direct (i.e., feeding on the lettuce leaves with 141Ce-contaminated surfaces) or an indirect/trophic (i.e., feeding on the lettuce leaves with systemically distributed 141Ce) route. To evaluate the influence of exposure routes, the Ce contents of the lettuce, snail tissues, and feces were determined by radioactivity measurements. The results show that both assimilation efficiencies (AEs) and food ingestion rates of Ce are greater for the trophic (indirect) exposure. The low AEs indicate that the CeO2 NPs ingested by snails were mostly excreted subsequently, and those that remained in the body were mainly concentrated in the digestive gland. XANES analysis shows that >85% of Ce was reduced to Ce(III) in the digestive gland under direct exposure, whereas Ce in the rest of the food chain (including feces) was largely in its original oxidized (IV) state. This study suggests that CeO2 NPs present in the environment may be taken up by producers and transferred to consumers along food chains and trophic transfer may affect food safety.


Dimensional analysis of the ventricle of an in situ perfused trout heart using echocardiography.
The linear dimensions of the ventricle of an in situ perfused trout heart were measured with an ultrasound scanner. Using a pyramidal model, linear dimensions taken from real-time, two-dimensional, echotomographic images of the ventricle were used to calculate ventricular end-diastolic and end-systolic volumes and, by the difference between them, stroke volumes for various filling and output pressures. Stroke volumes calculated from the ultrasound measurements were significantly correlated with actual stroke volumes determined from cardiac output and heart rate and had a regression line slope close to one. Increases in stroke volume with greater filling pressures resulted from increased ventricular end-diastolic volume with constant end-systolic volume; end-systolic volumes were negligible. Maintenance of stroke volume with increasing output pressure was by increases in both end-diastolic and end-systolic volumes. Decreases in stroke volume with very high output pressures may result from convergence of end-systolic and end-diastolic volumes.


Neurological evidence linguistic processes precede perceptual simulation in conceptual processing.
There is increasing evidence from response time experiments that language statistics and perceptual simulations both play a role in conceptual processing. In an EEG experiment we compared neural activity in cortical regions commonly associated with linguistic processing and visual perceptual processing to determine to what extent symbolic and embodied accounts of cognition applied. Participants were asked to determine the semantic relationship of word pairs (e.g., sky - ground) or to determine their iconic relationship (i.e., if the presentation of the pair matched their expected physical relationship). A linguistic bias was found toward the semantic judgment task and a perceptual bias was found toward the iconicity judgment task. More importantly, conceptual processing involved activation in brain regions associated with both linguistic and perceptual processes. When comparing the relative activation of linguistic cortical regions with perceptual cortical regions, the effect sizes for linguistic cortical regions were larger than those for the perceptual cortical regions early in a trial with the reverse being true later in a trial. These results map upon findings from other experimental literature and provide further evidence that processing of concept words relies both on language statistics and on perceptual simulations, whereby linguistic processes precede perceptual simulation processes.


Origin of ecdysis: fossil evidence from 535-million-year-old scalidophoran worms.
With millions of extant species, ecdysozoans (Scalidophora, Nematoida and Panarthropoda) constitute a major portion of present-day biodiversity. All ecdysozoans secrete an exoskeletal cuticle which must be moulted periodically and replaced by a larger one. Although moulting (ecdysis) has been recognized in early Palaeozoic panarthropods such as trilobites and basal groups such as anomalocaridids and lobopodians, the fossil record lacks clear evidence of ecdysis in early scalidophorans, largely because of difficulties in recognizing true exuviae. Here, we describe two types of exuviae in microscopic scalidophoran worms from the lowermost Cambrian Kuanchuanpu Formation ( ca 535 Ma) of China and reconstruct their moulting process. These basal scalidophorans moulted in a manner similar to that of extant priapulid worms, extricating themselves smoothly from their old tubular cuticle or turning their exuviae inside out like the finger of a glove. This is the oldest record of moulting in ecdysozoans. We also discuss the origin of ecdysis in the light of recent molecular analyses and the significance of moulting in the early evolution of animals.


Overexpression of NDRG1 is an indicator of poor prognosis in hepatocellular carcinoma.
Hepatocellular carcinoma is a highly lethal cancer that typically has poor prognosis. Prognostic markers can help in its clinical management and in understanding the biology of poor prognosis. Through an earlier gene expression study, we identified N-Myc downregulated gene 1 (NDRG1) to be significantly highly expressed in hepatocellular carcinoma compared to nontumor liver. As NDRG1 is a differentiation-related gene with putative metastasis suppressor activity, we investigated the clinical significance of its overexpression. Quantitative real-time polymerase chain reaction using an independent set of patient samples confirmed the significant overexpression of NDRG1 in hepatocellular carcinoma compared to nontumor liver samples (P<0.001). Additionally, high levels of NDRG1 transcript correlated with shorter overall survival (P<0.001), late tumor stage (P=0.001), vascular invasion (P=0.003), large tumor size (P=0.011), and high Edmondson-Steiner histological grade (P=0.005). Using immunohistochemistry, NDRG1 protein was found to be significantly overexpressed in hepatocellular carcinoma samples compared to nontumor liver or cirrhotic and benign liver lesions (P<0.001). Among the hepatocellular carcinoma samples, those which are moderately and poorly differentiated express higher levels of NDRG1 protein than those which are well-differentiated (P<0.005). Additionally, hepatocellular carcinomas with vascular invasion also express elevated levels of NDRG1 protein compared to those without vascular invasion (significant at P<0.005). Our results suggest NDRG1 to be a likely tumor marker for hepatocellular carcinoma, the overexpression of which is correlated with tumor differentiation, vascular invasion, and overall survival. Its significantly elevated expression in hepatocellular carcinoma could be a useful indicator of tumor aggressiveness and therefore patient prognosis.


Some characteristics of the resistance transfer factor (RTF) episome as determined by inactivation with tritium, P32, and gamma radiation.
The resistance transfer factor (RTF) episome was studied by measuring its inactivation by Co(60) gamma radiation, by incorporated P(32), and by tritium incorporated as tritium-labeled thymine. The D(37) for Co(60) irradiation was 7 to 9 x 10(4) rad. Growth of the bacteria harboring the RTF in BUdR (bromouracil deoxyriboside) increased the sensitivity of the RTF to the gamma radiation. The RTF was markedly inactivated by tritium after growth of the host (thymine requiring) bacteria in tritium-labeled thymine, thus further establishing the presence of thymine in the genome of the RTF. Assuming the efficiency of inactivation by P(32) to be 10%, the phosphorus content of the RTF was estimated to be about 2 x 10(5) P atoms/episome. The data suggest the RTF contains double stranded DNA with a molecular weight of the order of 3 to 8 x 10(7).


Integrative High Dimensional Multiple Testing with Heterogeneity under Data Sharing Constraints.
Identifying informative predictors in a high dimensional regression model is a critical step for association analysis and predictive modeling. Signal detection in the high dimensional setting often fails due to the limited sample size. One approach to improving power is through meta-analyzing multiple studies which address the same scientific question. However, integrative analysis of high dimensional data from multiple studies is challenging in the presence of between-study heterogeneity. The challenge is even more pronounced with additional data sharing constraints under which only summary data can be shared across different sites. In this paper, we propose a novel data shielding integrative large-scale testing (DSILT) approach to signal detection allowing between-study heterogeneity and not requiring the sharing of individual level data. Assuming the underlying high dimensional regression models of the data differ across studies yet share similar support, the proposed method incorporates proper integrative estimation and debiasing procedures to construct test statistics for the overall effects of specific covariates. We also develop a multiple testing procedure to identify significant effects while controlling the false discovery rate (FDR) and false discovery proportion (FDP). Theoretical comparisons of the new testing procedure with the ideal individual-level meta-analysis (ILMA) approach and other distributed inference methods are investigated. Simulation studies demonstrate that the proposed testing procedure performs well in both controlling false discovery and attaining power. The new method is applied to a real example detecting interaction effects of the genetic variants for statins and obesity on the risk for type II diabetes.


Evaluation of the infestation rate of Blaesoxipha alcedo in the carrion roller scarab Canthon cyanellus cyanellus and its effect on reproductive behavior.
This study documents Blaesoxipha alcedo (Aldrich; Diptera: Sarcophagidae) parasitizing the necrophagous ball roller beetle Canthon cyanellus cyanellus LeConte collected over 2 yr, and evaluates the reproductive behavior of parasitized beetles. Up to 52% of the beetles collected in the field had been parasitized and exhibited modified sexual behavior. In general, 21% of the males and 24% of the females had been parasitized. The number of parasitized males peaked in August 2000 (52%), but that of females peaked in May 2001 (52%). The lowest percentage of parasitized males (7%) was recorded in September 2000, June 2001, and July 2001; for females, parasitism was lowest in June 2000 (10%). The larva of this sarcophagid fly penetrates the host through the tegumentary membrane, leaving a scar. The larvae feed on the gonads of male and female C. c. cyanellus, castrating them, causing the loss of their ability to recognize individuals of the opposite sex, preventing reproduction, and lowering overall reproductive success.


Using a clinic based creativity initiative to reduce HIV related stigma at the Infectious Diseases Institute, Mulago National Referral Hospital, Uganda.
Stigma has been associated with chronic health conditions such as HIV/AIDS, leprosy, tuberculosis, Mental illness and Epilepsy. Different forms of stigma have been identified: enacted stigma, perceived stigma, and self stigma. Stigma is increasingly regarded as a key driver of the HIV/AIDS epidemic and has a major impact on public health interventions. The initiative was to provide activities in the clinic while patients waited to be seen by healthcare professionals. It was envisaged this would contribute to reduction of clinic based stigma felt by clients. This was a repeated cross-sectional survey (October-November 2005 and March-April 2007) that was conducted at the Infectious Diseases Institute clinic (IDC) at Mulago, the national referral hospital in Uganda. We utilized quantitative (survey) and qualitative (key informants, focus group discussions) methods to collect the data. Data were collected on stigma before the creativity initiative intervention was implemented, and a second phase survey was conducted to assess effectiveness of the interventions. Clients who attended the IDC before the creativity intervention were about twice as likely to fear catching an infection as those who came after the intervention. The proportion that had fears to be seen by a friend or relative at the clinic decreased. Thus during the implementation of the Creativity intervention, HIV related stigma was reduced in this clinic setting. The creativity intervention helped to build self esteem and improved communication among those attending the clinic; there was observed ambiance at the clinic and clients became empowered, with creative, communication and networking skills. Improved knowledge and communication are key in addressing self stigma among HIV positive individuals.


Breastfeeding Practices Among Women Living with HIV in KwaZulu-Natal, South Africa: An Observational Study.
Exclusive breastfeeding (EBF) is the safest infant feeding option in resource-limited settings, though women living with HIV have the lowest rates of EBF. Barriers to EBF in the absences of a formal intervention in women living with HIV in KwaZulu-Natal, where the prevalence of HIV among pregnant women is among the highest in the world, are understudied. Thus, this study sought to describe barriers to EBF and examine differences in social support, disclosure status, mood, and HIV-related stigma among women with different feeding methods. Women living with HIV enrolled in preventing mother-to-child transmission treatment (n = 156) were interviewed postpartum (M = 13.1 weeks) at a district hospital and self-reported infant feeding method, reasons not breastfeeding (if applicable), and HIV disclosure status. Mood, HIV-related stigma, functional social support, and HIV-related social support were also assessed. No participants reported mixed feeding, 30% reported EBF, and 70% reported exclusive formula feeding. Commonly reported reasons for not breastfeeding included fear of HIV transmission to the infant and being away from the infant for extended periods of time. Social support (p = 0.02) and HIV-related social support (p < 0.01) were significantly higher in women who had attempted breastfeeding compared to women who never attempted breastfeeding. Rates of EBF in this sample are lower than in other recent studies, suggesting this sample experiences multiple barriers to EBF. Healthcare providers should seek to correct misconceptions regarding HIV transmission and breastfeeding practices. Social and logistical support for EBF may be important considerations for future interventions.


Antibody response is reduced following vaccination with 7-valent conjugate pneumococcal vaccine in adult methotrexate-treated patients with established arthritis, but not those treated with tumor necrosis factor inhibitors.
To study the influence of antiinflammatory treatments, including methotrexate (MTX) and tumor necrosis factor (TNF) inhibitors, on antibody response following vaccination using a 7-valent conjugate pneumococcal vaccine in adult patients with established arthritis. Patients with rheumatoid arthritis (RA) or spondylarthropathy (SpA) (including psoriatic arthritis) were vaccinated (n = 505). All patients were stratified into 6 prespecified groups based on diagnosis and treatment (RA patients receiving MTX, RA patients receiving anti-TNF agents and MTX, RA patients receiving TNF inhibitors as monotherapy, SpA patients receiving anti-TNF agents and MTX, SpA patients receiving TNF inhibitors as monotherapy, and SpA patients receiving nonsteroidal antiinflammatory drugs [NSAIDs] and/or analgesics). SpA patients receiving only NSAIDs/analgesics served as a control group. All patients received 1 dose (0.5 ml) of vaccine intramuscularly. Levels of IgG antibodies against 23F and 6B serotypes were measured at vaccination and at 4-6 weeks following vaccination, using standardized enzyme-linked immunosorbent assays. Positive antibody response was defined as an antibody response ratio (ARR) (i.e., ratio of post- to prevaccination antibody levels) of ≥2. The ARR differed significantly between the groups. A better ARR was seen among patients in the control group compared to those in groups treated with MTX or MTX in combination with TNF inhibitors. Among patients treated with TNF inhibitors as monotherapy, ARRs for both serotypes were lower numerically, but were not significantly different, compared to those in controls. Ongoing MTX treatment was predictive of reduced response (odds ratio 0.41 [95% confidence interval 0.24-0.68], P = 0.001). Higher age was associated with impaired positive antibody response. Concomitant prednisolone treatment elicited better positive antibody response in patients with RA. Treatment with MTX and higher age were predictive of an impaired antibody response to the 7-valent conjugate pneumococcal vaccine in this cohort of patients with chronic arthritis. TNF inhibitors did not significantly affect antibody responses.


edn1 and hand2 Interact in early regulation of pharyngeal arch outgrowth during zebrafish development.
Endothelin-1 (Edn1) signaling provides a critical input to development of the embryonic pharygneal arches and their skeletal derivatives, particularly the articulating joints and the ventral skeleton including the lower jaw. Previous work in zebrafish has mostly focused on the role of Edn1 in dorsal-ventral (DV) patterning, but Edn1 signaling must also regulate tissue size, for with severe loss of the pathway the ventral skeleton is not only mispatterned, but is also prominently hypoplastic--reduced in size. Here we use mutational analyses to show that in the early pharyngeal arches, ventral-specific edn1-mediated proliferation of neural crest derived cells is required for DV expansion and outgrowth, and that this positive regulation is counterbalanced by a negative one exerted through a pivotal, ventrally expressed Edn1-target gene, hand2. We also describe a new morphogenetic cell movement in the ventral first arch, sweeping cells anterior in the arch to the region where the lower jaw forms. This movement is negatively regulated by hand2 in an apparently edn1-independent fashion. These findings point to complexity of regulation by edn1 and hand2 at the earliest stages of pharyngeal arch development, in which control of growth and morphogenesis can be genetically separated.


Hyperglycemia augments ischemic brain injury: in vivo MR imaging/spectroscopic study with nicardipine in cats with occluded middle cerebral arteries.
Hyperglycemia is often associated with an increased frequency of cerebrovascular disease and exacerbation of neuronal injury in focal ischemic cerebral infarction. We used a combination of high-field proton MR imaging and 1H and 31P MR spectroscopy to investigate whether hyperglycemia would adversely influence cerebral metabolism and eventual infarct size following unilateral occlusion of the middle cerebral artery (MCA) of cats pretreated with the calcium channel blocker nicardipine. Normoglycemic animals injected with 10 micrograms/kg of nicardipine (8 micrograms.kg-1.hr-1 maintenance dose) manifested only mild disturbances in phosphorus metabolism and cerebral pH regulation compared with untreated controls, and showed a significant reduction in infarct size 7 hr after MCA occlusion. By comparison, hyperglycemic cats (plasma glucose, 200-300 mg/dl) had significantly reduced cerebral high-energy phosphates, elevated lactic acid, and larger ischemic lesions in the occluded MCA territory, irrespective of whether they were treated with nicardipine. These results indicate that moderate hyperglycemia can exaggerate ischemic brain damage by enhancing formation of tissue lactic acid and impairing normal phosphorus metabolism. One implication of this study is that dextrose should not be provided to patients with acute ischemic stroke.


Suppressive effects of cannabidiol on antigen-specific antibody production and functional activity of splenocytes in ovalbumin-sensitized BALB/c mice.
Cannabidiol (CBD) and cannabis-based medicines are potential therapeutic agents. Because the immune system has been widely demonstrated to be affected by psychoactive cannabinoids, such as Delta(9)-tetrahydrocannabinol, the objective of the present studies is to investigate the immunomodulatory effect of CBD, the major non-psychoactive cannabinoid in marijuana. BALB/c mice were intraperitoneally administered with a single dose of CBD (5-20 mg/kg) prior to ovalbumin (OVA) sensitization, and the serum production of antigen-specific antibodies was measured 7 days post OVA sensitization. The serum level of OVA-specific IgM was significantly attenuated by a high dose of CBD (20 mg/kg), and OVA-specific IgG(1) and IgG(2a) by all 3 doses of CBD. Concordantly, splenocytes of mice administered with CBD (5 or 20 mg/kg) produced less IL-2, IL-4 and IFN-gamma than those of vehicle-treated controls, upon ex vivo stimulation with phorbol ester plus calcium ionophore. Likewise, T-cell mitogen (concanavalin A)-induced proliferation of splenocytes was also markedly suppressed in mice administered with CBD. Furthermore, the observed ex vivo effects of CBD on cytokine production and T-cell proliferation were confirmed in splenocytes directly exposed to CBD (1-8 microM) in vitro, indicating a direct effect by CBD. Taken together, the results demonstrated that CBD markedly suppressed antigen-specific antibody production in OVA-sensitized mice, and suggest that CBD-mediated suppression of humoral immunity could be mediated by the impaired functions of splenocytes.


Effects of marijuana use on impulsivity and hostility in daily life.
Marijuana use is increasingly prevalent among young adults. While research has found adverse effects associated with marijuana use within experimentally controlled laboratory settings, it is unclear how recreational marijuana use affects day-to-day experiences in users. The present study sought to examine the effects of marijuana use on within-person changes in impulsivity and interpersonal hostility in daily life using smartphone administered assessments. Forty-three participants with no substance dependence reported on their alcohol consumption, tobacco use, recreational marijuana use, impulsivity, and interpersonal hostility over the course of 14 days. Responses were analyzed using multilevel modeling. Marijuana use was associated with increased impulsivity on the same day and the following day relative to days when marijuana was not used, independent of alcohol use. Marijuana was also associated with increased hostile behaviors and perceptions of hostility in others on the same day when compared to days when marijuana was not used. These effects were independent of frequency of marijuana use or alcohol use. There were no significant effects of alcohol consumption on impulsivity or interpersonal hostility. Marijuana use is associated with changes in impulse control and hostility in daily life. This may be one route by which deleterious effects of marijuana are observed for mental health and psychosocial functioning. Given the increasing prevalence of recreational marijuana use and the potential legalization in some states, further research on the potential consequences of marijuana use in young adults' day-to-day life is warranted.


Epidemiology of COVID-19 infections on routine polymerase chain reaction (PCR) and serology testing in Coastal Kenya.
Background: There are limited studies in Africa describing the epidemiology, clinical characteristics and serostatus of individuals tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We tested routine samples from the Coastal part of Kenya between 17 th March 2020 and 30 th June 2021. Methods: SARS-CoV-2 infections identified using reverse transcription polymerase chain reaction (RT-PCR) and clinical surveillance data at the point of sample collection were used to classify as either symptomatic or asymptomatic. IgG antibodies were measured in sera samples, using a well validated in-house enzyme-linked immunosorbent assay (ELISA). Results: Mombasa accounted for 56.2% of all the 99,694 naso-pharyngeal/oro-pharyngeal swabs tested, and males constituted the majority tested (73.4%). A total of 7737 (7.7%) individuals were SARS-CoV-2 positive by RT-PCR. The majority (i.e., 92.4%) of the RT-PCR positive individuals were asymptomatic. Testing was dominated by mass screening and travellers, and even at health facility level 91.6% of tests were from individuals without symptoms. Out of the 97,124 tests from asymptomatic individuals 7,149 (7%) were positive and of the 2,568 symptomatic individuals 588 (23%) were positive. In total, 2458 serum samples were submitted with paired naso-pharyngeal/oro-pharyngeal samples and 45% of the RT-PCR positive samples and 20% of the RT-PCR negative samples were paired with positive serum samples. Symptomatic individuals had significantly higher antibody levels than asymptomatic individuals and become RT-PCR negative on repeat testing earlier than asymptomatic individuals. Conclusions: In conclusion, the majority of SARS-CoV-2 infections identified by routine testing in Coastal Kenya were asymptomatic. This reflects the testing practice of health services in Kenya, but also implies that asymptomatic infection is very common in the population. Symptomatic infection may be less common, or it may be that individuals do not present for testing when they have symptoms.


NPM1 histone chaperone is upregulated in glioblastoma to promote cell survival and maintain nucleolar shape.
Glioblastoma (grade IV glioma) is the most common and aggressive adult brain tumor. A better understanding of the biology of glioblastoma cells is crucial to identify molecular targets stimulating cell death. NPM1 (nucleophosmin) is a multifunctional chaperone that plays an important role in cancer development. Herein, NPM1 was analyzed by immunohistochemistry in human astrocytic gliomas. NPM1 was detected in all tumors but with a significantly higher staining intensity in grade IV than in low grade tumors. Depletion of NPM1 had only modest effects on the viability of U251MG, U1242MG, and U343MGa Cl2:6 glioma cells, despite alterations in nucleolar morphology. Glioma cell cultures depleted of NPM1 exposed to micromolar levels of actinomycin D were more prone to cell death (apoptosis) compared to cultures retaining NPM1. We had previously found that NPM1 binds to linker histone H1.5. Here we could show that silencing of H1.5 triggered glioma cell apoptosis as evidenced by a marked increase in both the numbers of cleaved caspase-3(+) cells and in the amounts of cleaved PARP. Enforced expression of NPM1 suppressed apoptosis in H1.5 depleted glioma cells. Although our studies would suggest little effectiveness of targeting NPM1 alone there could be potential using it as a combination treatment.


Microbial population analysis of the salivary glands of ticks; a possible strategy for the surveillance of bacterial pathogens.
Ticks are one of the most important blood-sucking vectors for infectious microorganisms in humans and animals. When feeding they inject saliva, containing microbes, into the host to facilitate the uptake of blood. An understanding of the microbial populations within their salivary glands would provide a valuable insight when evaluating the vectorial capacity of ticks. Three tick species (Ixodes ovatus, I. persulcatus and Haemaphysalis flava) were collected in Shizuoka Prefecture of Japan between 2008 and 2011. Each tick was dissected and the salivary glands removed. Bacterial communities in each salivary gland were characterized by 16S amplicon pyrosequencing using a 454 GS-Junior Next Generation Sequencer. The Ribosomal Database Project (RDP) Classifier was used to classify sequence reads at the genus level. The composition of the microbial populations of each tick species were assessed by principal component analysis (PCA) using the Metagenomics RAST (MG-RAST) metagenomic analysis tool. Rickettsia-specific PCR was used for the characterization of rickettsial species. Almost full length of 16S rDNA was amplified in order to characterize unclassified bacterial sequences obtained in I. persulcatus female samples. The numbers of bacterial genera identified for the tick species were 71 (I. ovatus), 127 (I. persulcatus) and 59 (H. flava). Eighteen bacterial genera were commonly detected in all tick species. The predominant bacterial genus observed in all tick species was Coxiella. Spiroplasma was detected in Ixodes, and not in H. flava. PCA revealed that microbial populations in tick salivary glands were different between tick species, indicating that host specificities may play an important role in determining the microbial complement. Four female I. persulcatus samples contained a high abundance of several sequences belonging to Alphaproteobacteria symbionts. This study revealed the microbial populations within the salivary glands of three species of ticks, and the results will contribute to the knowledge and prediction of emerging tick-borne diseases.


Characterizing ingestive behavior through licking microstructure: Underlying neurobiology and its use in the study of obesity in animal models.
Ingestive behavior is controlled by multiple distinct peripheral and central physiological mechanisms that ultimately determine whether a particular food should be accepted or avoided. As rodents consume a fluid they display stereotyped rhythmic tongue movements, and by analyzing the temporal distribution of pauses of licking, it is possible through analyses of licking microstructure to uncover dissociable evaluative and motivational variables that contribute to ingestive behavior. The mean number of licks occurring within each burst of licking (burst and cluster size) reflects the palatability of the consumed solution, whereas the frequency of initiating novel bouts of licking behavior (burst and cluster number) is dependent upon the degree of gastrointestinal inhibition that accrues through continued fluid ingestion. This review describes the analysis of these measures within a context of the behavioral variables that come to influence the acceptance or avoidance of a fluid, and the neurobiological mechanisms that underlie alterations in the temporal distribution of pauses of licks. The application of these studies to models of obesity in animals is also described.


RNAi depleted Drosophila cell extracts to dissect signaling pathways leading to actin polymerization.
Dissection of signal transduction pathways leading to actin polymerization has been performed in cytosolic extracts. In such assays, the implication of an effector molecule is demonstrated by the loss of actin polymerization upon its depletion and the restoration of actin polymerization upon its add-back. Two major limitations in the wide use of this approach have been the availability of immunodepleting antibodies and the functional redundancy for many classes of effector molecules encoded by vertebrate genomes. To circumvent these limitations, we developed extracts derived from S2 Drosophila cells, which are competent for actin polymerization. In this system, depleted extracts are simply obtained from cells cultured with long double stranded RNAs in the medium. We validated the method by showing that beads coated with the C-terminal domain of Wave2 were no longer able to trigger actin polymerization in an extract depleted of the Arp2/3 complex. We also examined the complete set of Drosophila small GTPases of the Rho family for their ability to polymerize actin in such extracts, and found that only dCdc42 was able to induce actin polymerization. Using RNAi depleted extract, we confirmed that dCdc42 triggers actin polymerization in a Wasp dependent manner.


The Use of Retinoids for the Prevention and Treatment of Skin Cancers: An Updated Review.
Retinoids are natural and synthetic vitamin A derivatives that are effective for the prevention and the treatment of non-melanoma skin cancers (NMSC). NMSCs constitute a heterogenous group of non-melanocyte-derived skin cancers that impose substantial burdens on patients and healthcare systems. They include entities such as basal cell carcinoma and cutaneous squamous cell carcinoma (collectively called keratinocyte carcinomas), cutaneous lymphomas and Kaposi's sarcoma among others. The retinoid signaling pathway plays influential roles in skin physiology and pathology. These compounds regulate diverse biological processes within the skin, including proliferation, differentiation, angiogenesis and immune regulation. Collectively, retinoids can suppress skin carcinogenesis. Both topical and systemic retinoids have been investigated in clinical trials as NMSC prophylactics and treatments. Desirable efficacy and tolerability in clinical trials have prompted health regulatory bodies to approve the use of retinoids for NMSC management. Acceptable off-label uses of these compounds as drugs for skin cancers are also described. This review is a comprehensive outline on the biochemistry of retinoids, their activities in the skin, their effects on cancer cells and their adoption in clinical practice.


A cluster RCT to improve workplace mental health in a policing context: Findings of a mixed-methods implementation evaluation.
We conducted a cluster randomized trial of a workplace mental health intervention in an Australian police department. The intervention was co-designed and co-implemented with the police department. Intervention elements included tailored mental health literacy training for all members of participating police stations, and a leadership development and coaching program for station leaders. This study presents the results of a mixed-methods implementation evaluation of the trial. Descriptive quantitative analyses characterized the extent of participation in intervention activities, complemented by a qualitative descriptive analysis of transcripts of 60 semistructured interviews with 53 persons and research team field notes. Participation rates in the multicomponent leadership development activities were highly variable, ranging from <10% to approximately 60% across stations. Approximately 50% of leaders and <50% of troops completed the mental health literacy training component of the intervention. Barriers to implementation included rostering challenges, high staff turnover and changes, competing work commitments, staff shortages, limited internal personnel resources to deliver the mental health literacy training, organizational cynicism, confidentiality concerns, and limited communication about the intervention by station command or station champions. Facilitators of participation were also identified, including perceived need for and benefits of the intervention, engagement at various levels, the research team's ability to create buy-in and manage stakeholder relationships, and the use of external, credible leadership development coaches. Implementation fell far short of expectations. The identified barriers and facilitators should be considered in the design and implementation of similar workplace mental health interventions.


Elucidating the function of non catalytic domains of collagenases and aggrecanases.
Metalloproteinases that degrade extracellular matrix molecules play important roles in development and progression of various diseases. Among them, collagenases are unique as they have an ability to degrade triple helical interstitial collagens into 3/4 and 1/4 fragments, a crucial step for collagenolysis in the tissue. Collagenases, consisting of a catalytic domain and a hemopexin domain, requires both domains for collagenolysis. The enzymes unwind triple helical collagen before they hydrolyze the peptide bonds. Aggrecanases are also multidomain metalloproteinases belonging to the ADAMTS family, and the noncatalytic ancillary domains also play an important role in recognition of aggrecan and their activities. Attenuation of collagenase and aggrecanase activities will be achieved by inhibitors or antibodies that interact directly with those noncatalytic ancillary domains (exosite inhibitors). Such molecules will be attractive for therapy as they will be highly selective because they are based on the unique mechanism of each proteinase.


Lymphedema: Improving Screening and Treatment Among At-Risk Breast Cancer Survivors.
Inadequate peripheral lymphedema screening is a persistent gap in the provision of quality survivorship care at many breast cancer centers. Lymphedema should be identified and treated during the subclinical stage, when it is more likely reversible. The purpose of this project was to integrate a screening process for patients with breast cancer at risk for lymphedema, using bioimpedance spectroscopy via the L-Dex®, in a breast cancer center. A protocol was developed that included specific criteria for early patient referral to physical therapy.


Multi-Omics Integration Highlights the Role of Ubiquitination in CCl4-Induced Liver Fibrosis.
Liver fibrosis is the excessive accumulation of extracellular matrix proteins that occurs in chronic liver disease. Ubiquitination is a post-translational modification that is crucial for a plethora of physiological processes. Even though the ubiquitin system has been implicated in several human diseases, the role of ubiquitination in liver fibrosis remains poorly understood. Here, multi-omics approaches were used to address this. Untargeted metabolomics showed that carbon tetrachloride (CCl4)-induced liver fibrosis promotes changes in the hepatic metabolome, specifically in glycerophospholipids and sphingolipids. Gene ontology analysis of public deposited gene array-based data and validation in our mouse model showed that the biological process "protein polyubiquitination" is enriched after CCl4-induced liver fibrosis. Finally, by using transgenic mice expressing biotinylated ubiquitin (bioUb mice), the ubiquitinated proteome was isolated and characterized by mass spectrometry in order to unravel the hepatic ubiquitinated proteome fingerprint in CCl4-induced liver fibrosis. Under these conditions, ubiquitination appears to be involved in the regulation of cell death and survival, cell function, lipid metabolism, and DNA repair. Finally, ubiquitination of proliferating cell nuclear antigen (PCNA) is induced during CCl4-induced liver fibrosis and associated with the DNA damage response (DDR). Overall, hepatic ubiquitome profiling can highlight new therapeutic targets for the clinical management of liver fibrosis.


n-3 Fatty acids inhibit transcription of human IL-13: implications for development of T helper type 2 immune responses.
Fish oil supplementation during pregnancy has been associated with lower levels of cord blood IL-13, suggesting that the administration of n-3 fatty acids may attenuate the development of allergic disease. The present study aimed to investigate the mechanism by which n-3 fatty acid administration influences the production of IL-13. Pregnant BALB/c mice were fed nutritionally complete high-fat diets (15 %, w/w) with an n-3 fatty acid-enriched (DHA 1 %, w/w) or control diet (0 % DHA) immediately following delivery. Pups were exposed during suckling and weaned to the maternal diet for the remainder of the study. The production of IL-13, IL-4, IL-10 and interferon-γ from the splenocytes of ovalbumin (ova)-sensitised animals was assessed following in vitro ova stimulation or unstimulated conditions. Human T helper type 2 (Th2) cells were mitogen-stimulated in the presence or absence of DHA (10 μM) and assessed for IL-13 and IL-4 expression using intracellular flow cytometry. The influence on transcriptional activation was studied using a human IL-13 promoter reporter construct and electromobility shift assay. Ova-activated splenocytes from DHA-fed mice produced less IL-13 (57.2 (se 21.7) pg/ml) and IL-4 (7.33 (SE 3.4) pg/ml) compared with cells from the animals fed the control diet (161.5 (SE 45.0), P< 0.05; 33.2 (SE 11.8), P< 0.05). In vitro, DHA inhibited the expression of IL-13 protein from human Th2 cells as well as transcriptional activation and binding of the transcription factors cyclic AMP response element binding and activating transcription factor 2 to the human IL-13 promoter. These data indicate the potential of n-3 fatty acids to attenuate IL-13 expression, and suggest that they may subsequently reduce allergic sensitisation and the development of allergic disease.


Current trends in the treatment of polycystic ovary syndrome with desire for children.
Polycystic ovary syndrome (PCOS), one of the most frequent endocrine diseases, affects approximately 5%-10% of women of childbearing age and constitutes the most common cause of female sterility regardless of the need or not for treatment, a change in lifestyle is essential for the treatment to work and ovulation to be restored. Obesity is the principal reason for modifying lifestyle since its reduction improves ovulation and the capacity for pregnancy and lowers the risk of miscarriage and later complications that may occur during pregnancy (gestational diabetes, pre-eclampsia, etc). When lifestyle modification is not sufficient, the first step in ovulation induction is clomiphene citrate. The second-step recommendation is either exogenous gonadotrophins or laparoscopic ovarian surgery. Recommended third-line treatment is in vitro fertilization. Metformin use in PCOS should be restricted to women with glucose intolerance.


Fish in habitats with higher motorboat disturbance show reduced sensitivity to motorboat noise.
Anthropogenic noise can negatively impact many taxa worldwide. It is possible that in noisy, high-disturbance environments, the range and severity of impacts could diminish over time, but the influence of previous disturbance remains untested in natural conditions. This study demonstrates the effects of motorboat noise on the physiology of an endemic cichlid fish in Lake Malawi. Exposure to motorboats (driven 20-100 m from fish) and loudspeaker playback of motorboat noise both elevated the oxygen-consumption rate at a single lower-disturbance site, characterized by low historic and current motorboat activity. Repeating this assay at further lower-disturbance sites revealed a consistent effect of elevated oxygen consumption in response to motorboat disturbance. However, when similar trials were repeated at four higher-disturbance sites, no effect of motorboat exposure was detected. These results demonstrate that disturbance history can affect local population responses to noise. Action regarding noise pollution should consider the past, as well as the present, when planning for the future.