Disruption of cytochrome P4501A2 in mice leads to increased susceptibility to hyperoxic lung injury.
Hyperoxia contributes to acute lung injury in diseases such as acute respiratory distress syndrome. Cytochrome P450 (CYP) 1A enzymes have been implicated in hyperoxic lung injury, but the mechanistic role of CYP1A2 in pulmonary injury is not known. We hypothesized that mice lacking the gene Cyp1a2 (which is predominantly expressed in the liver) will be more sensitive to lung injury and inflammation mediated by hyperoxia and that CYP1A2 will play a protective role by attenuating lipid peroxidation and oxidative stress in the lung. Eight- to ten-week-old WT (C57BL/6) or Cyp1a2(-/-) mice were exposed to hyperoxia (>95% O2) or maintained in room air for 24-72 h. Lung injury was assessed by determining the ratio of lung weight/body weight (LW/BW) and by histology. Extent of inflammation was determined by measuring the number of neutrophils in the lung as well as cytokine expression. The Cyp1a2(-/-) mice under hyperoxic conditions showed increased LW/BW ratios, lung injury, neutrophil infiltration, and IL-6 and TNF-α levels and augmented lipid peroxidation, as evidenced by increased formation of malondialdehyde- and 4-hydroxynonenal-protein adducts and pulmonary isofurans compared to WT mice. In vitro experiments showed that the F2-isoprostane PGF2-α is metabolized by CYP1A2 to a dinor metabolite, providing evidence for a catalytic role for CYP1A2 in the metabolism of F2-isoprostanes. In summary, our results support the hypothesis that hepatic CYP1A2 plays a critical role in the attenuation of hyperoxic lung injury by decreasing lipid peroxidation and oxidative stress in vivo.


Optical add-drop filters based on photonic crystal ring resonators.
We present here an optical add-drop filter (ADF) design based on ultra-compact photonic crystal ring resonators (PCRRs). The normalized transmission spectra for single-ring and dual-ring configurations have been investigated by using the two-dimensional finite-difference time-domain (FDTD) technique in a square lattice dielectric-rod photonic-crystal structure. With the introduction of four scatterers at the corners of quasisquare- ring PCRR, high wavelength selectivity and close to 100% drop efficiency can be obtained. Both backward- and forward-dropping were achieved by controlling the coupling efficiency between two PCRR rings for resonant modes with different symmetry. The resonant-mode quality factor Q and the wavelength tunability were also analyzed, opening opportunities for PCRRs as ultra-compact filters, optical add-drop multiplexers, electrooptical N x N switches and electrooptical modulators.


Formation of glasses from liquids and biopolymers.
Glasses can be formed by many routes. In some cases, distinct polyamorphic forms are found. The normal mode of glass formation is cooling of a viscous liquid. Liquid behavior during cooling is classified between "strong" and "fragile," and the three canonical characteristics of relaxing liquids are correlated through the fragility. Strong liquids become fragile liquids on compression. In some cases, such conversions occur during cooling by a weak first-order transition. This behavior can be related to the polymorphism in a glass state through a recent simple modification of the van der Waals model for tetrahedrally bonded liquids. The sudden loss of some liquid degrees of freedom through such first-order transitions is suggestive of the polyamorphic transition between native and denatured hydrated proteins, which can be interpreted as single-chain glass-forming polymers plasticized by water and cross-linked by hydrogen bonds. The onset of a sharp change in d<r(2)>dT(<r(2)> is the Debye-Waller factor and T is temperature) in proteins, which is controversially indentified with the glass transition in liquids, is shown to be general for glass formers and observable in computer simulations of strong and fragile ionic liquids, where it proves to be close to the experimental glass transition temperature. The latter may originate in strong anharmonicity in modes ("bosons"), which permits the system to access multiple minima of its configuration space. These modes, the Kauzmann temperature T(K), and the fragility of the liquid, may thus be connected.


Prospective Correlation of Magnetic Resonance Tumor Regression Grade With Pathologic Outcomes in Total Neoadjuvant Therapy for Rectal Adenocarcinoma.
Total neoadjuvant therapy (TNT) is a newly established standard treatment for rectal adenocarcinoma. Current methods to communicate magnitudes of regression during TNT are subjective and imprecise. Magnetic resonance tumor regression grade (MR-TRG) is an existing, but rarely used, regression grading system. Prospective validation of MR-TRG correlation with pathologic response in patients undergoing TNT is lacking. Utility of adding diffusion-weighted imaging to MR-TRG is also unknown. We conducted a multi-institutional prospective imaging substudy within NRG-GI002 (ClinicalTrials.gov identifier: NCT02921256) examining the ability of MR-based imaging to predict pathologic complete response (pCR) and correlate MR-TRG with the pathologic neoadjuvant response score (NAR). Serial MRIs were needed from 110 patients. Three radiologists independently, then collectively, reviewed each MRI for complete response (mriCR), which was tested for positive predictive value (PPV), negative predictive value (NPV), sensitivity, and specificity with pCR. MR-TRG was examined for association with the pathologic NAR score. All team members were blinded to pathologic data. A total of 121 patients from 71 institutions met criteria: 28% were female (n = 34), 84% White (n = 101), and median age was 55 (24-78 years). Kappa scores for T- and N-stage after TNT were 0.38 and 0.88, reflecting fair agreement and near-perfect agreement, respectively. Calling an mriCR resulted in a kappa score of 0.82 after chemotherapy and 0.56 after TNT reflected near-perfect agreement and moderate agreement, respectively. MR-TRG scores were associated with pCR (P < .01) and NAR (P < .0001), PPV for pCR was 40% (95% CI, 26 to 53), and NPV was 84% (95% CI, 75 to 94). MRI alone is a poor tool to distinguish pCR in rectal adenocarcinoma undergoing TNT. However, the MR-TRG score presents a now validated method, correlated with pathologic NAR, which can objectively measure regression magnitude during TNT.


Low is large: spatial location and pitch interact in voice-based body size estimation.
The binding of incongruent cues poses a challenge for multimodal perception. Indeed, although taller objects emit sounds from higher elevations, low-pitched sounds are perceptually mapped both to large size and to low elevation. In the present study, we examined how these incongruent vertical spatial cues (up is more) and pitch cues (low is large) to size interact, and whether similar biases influence size perception along the horizontal axis. In Experiment 1, we measured listeners' voice-based judgments of human body size using pitch-manipulated voices projected from a high versus a low, and a right versus a left, spatial location. Listeners associated low spatial locations with largeness for lowered-pitch but not for raised-pitch voices, demonstrating that pitch overrode vertical-elevation cues. Listeners associated rightward spatial locations with largeness, regardless of voice pitch. In Experiment 2, listeners performed the task while sitting or standing, allowing us to examine self-referential cues to elevation in size estimation. Listeners associated vertically low and rightward spatial cues with largeness more for lowered- than for raised-pitch voices. These correspondences were robust to sex (of both the voice and the listener) and head elevation (standing or sitting); however, horizontal correspondences were amplified when participants stood. Moreover, when participants were standing, their judgments of how much larger men's voices sounded than women's increased when the voices were projected from the low speaker. Our results provide novel evidence for a multidimensional spatial mapping of pitch that is generalizable to human voices and that affects performance in an indirect, ecologically relevant spatial task (body size estimation). These findings suggest that crossmodal pitch correspondences evoke both low-level and higher-level cognitive processes.


Uptake of a 21-gene expression assay in breast cancer practice: views of academic and community-based oncologists.
Advances in personalized medicine have produced novel tests and treatment options for women with breast cancer. Relatively little is known about the process by which such tests are adopted into oncology practice. The objectives of the present study were to understand the experiences of medical oncologists with multigene expression profile (gep) tests, including their adoption into practice in early-stage breast cancer, and the perceptions of the oncologists about the influence of test results on treatment decision-making. We conducted a qualitative descriptive study involving interviews with medical oncologists from academic and community cancer centres or hospitals in 8 communities in Ontario. A 21-gene breast cancer assay was used as the example of gep testing. Qualitative analytic techniques were used to identify the main themes. Of 28 oncologists who were approached, 21 (75%) participated in the study [median age: 43 years; 12 women (57%)]. Awareness and knowledge of gep testing were derived from several sources: international scientific meetings, participation in clinical studies, discussions with respected colleagues, and manufacturer-sponsored meetings. Oncologists observed that incorporating gep testing into their clinical practice resulted in several changes, including longer consultation times, second visits, and taking steps to minimize treatment delays. Oncologists expressed divergent opinions about the strength of evidence and added value of gep testing in guiding treatment decisions. Incorporation of gep testing into clinical practice in early-stage breast cancer required oncologists to make changes to their usual routines. The opinions of oncologists about the quality of evidence underpinning the test affected how much weight they gave to test results in treatment decision-making.


The human ERG1 channel polymorphism, K897T, creates a phosphorylation site that inhibits channel activity.
Single-nucleotide polymorphisms (SNPs) in the human ether-a-go-go-related gene 1, hERG1, are associated with cardiac arrhythmias. The Kv11.1 channels encoded by hERG1 are also essential for rhythmic excitability of the pituitary, where they are regulated by thyroid hormone through a signal transduction cascade involving the phosphatidylinositol 3-kinase (PI3K) and the Ser/Thr-directed protein phosphatase, PP5. Here, we show that the hERG1 polymorphism at codon 897, which is read as a Thr instead of a Lys, creates a phosphorylation site for the Akt protein kinase on the Kv11.1 channel protein. Consequently, hormonal signaling through the PI3K signaling cascade, which normally stimulates K897 channels through PP5-mediated dephosphorylation, inhibits T897 channels through Akt-mediated phosphorylation. Thus, hormonal regulation of Kv11.1 in humans with the T897 polymorphism is predicted to prolong the QT interval of cardiac myocytes. A systematic bioinformatics search for SNPs in human ion channel genes identified 15 additional candidates for such "phosphorylopathies," which are predicted to create or destroy putative phosphorylation sites. Changes in protein phosphorylation might represent a general mechanism for the interaction of genetic variation and environment on human health.


Multidisciplinary care for Guillain-Barré syndrome.
Guillain-Barré syndrome is a significant cause of new long-term disability, which is thought to be amenable to multidisciplinary care, but the evidence base for its effectiveness is unclear. To assess the effectiveness of multidisciplinary care in adults with Guillain-Barré syndrome, especially the types of approaches that are effective (settings, intensity) and the outcomes that are affected. We searched the Cochrane Neuromuscular Disease Group Specialized Register (20 May 2010), The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library 2010, Issue 2), MEDLINE (1966 to May 2010), EMBASE (1980 to May 2010), CINAHL (1982 to May 2010), AMED (1985 to May 2010), PEDro (1982 to May 2010) and LILACS (1982 to May 2010). We checked the bibliographies of papers identified and contacted their authors and known experts in the field seeking published and unpublished trials. Randomised and controlled clinical trials that compared multidisciplinary care in Guillain-Barré syndrome with either routinely available local services or lower levels of intervention, or studies that compared multidisciplinary care in different settings or at different levels of intensity.Studies of 'other designs' (such as observational studies) were included only in the Discussion since such studies could only be of limited contribution to the best evidence synthesis. We performed a 'best evidence' synthesis based on methodological quality. No randomised controlled trials or controlled clinical trials were identified. We summarised the results of three observational studies in the Discussion section of this review. In the absence of randomised controlled trials or controlled clinical trials, the 'best' evidence to date comes from three 'very low quality' observational studies. These provide some support for improved disability in the short term (less than 6 months) with high intensity inpatient multidisciplinary rehabilitation; and for improved quality of life, as measured by a reduction in handicap (participation). These conclusions are tentative and the gap in current research should not be interpreted as proof that multidisciplinary care is ineffective. Further research is needed into appropriate study designs; outcome measurement; caregiver needs; and the evaluation of optimal settings, type, intensity or frequency and cost-effectiveness of multidisciplinary care in the Guillain-Barré syndrome population.


Mapping the operational landscape of microRNAs in synthetic gene circuits.
MicroRNAs are a class of short, noncoding RNAs that are ubiquitous modulators of gene expression, with roles in development, homeostasis, and disease. Engineered microRNAs are now frequently used as regulatory modules in synthetic biology. Moreover, synthetic gene circuits equipped with engineered microRNA targets with perfect complementarity to endogenous microRNAs establish an interface with the endogenous milieu at the single-cell level. The function of engineered microRNAs and sensor systems is typically optimized through extensive trial-and-error. Here, using a combination of synthetic biology experimentation in human embryonic kidney cells and quantitative analysis, we investigate the relationship between input genetic template abundance, microRNA concentration, and output under microRNA control. We provide a framework that employs the complete operational landscape of a synthetic gene circuit and enables the stepwise development of mathematical models. We derive a phenomenological model that recapitulates experimentally observed nonlinearities and contains features that provide insight into the microRNA function at various abundances. Our work facilitates the characterization and engineering of multi-component genetic circuits and specifically points to new insights on the operation of microRNAs as mediators of endogenous information and regulators of gene expression in synthetic biology.


Phosphorylated Akt Expression as a Favorable Prognostic Factor for Patients Undergoing Curative Resection and Adjuvant Chemoradiotherapy for Proximal Extrahepatic Bile Duct Cancer.
To evaluate the prognostic significance of phosphorylated Akt (p-Akt), phosphorylated mammalian target of rapamycin (p-mTOR), and total phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expressions in patients undergoing adjuvant chemoradiotherapy (CRT) for proximal extrahepatic bile duct (EHBD) cancer. Sixty-three patients with proximal EHBD cancer who underwent curative resection followed by adjuvant CRT were enrolled into this study. Postoperative radiotherapy was delivered to tumor bed and regional lymph nodes up to a median of 40 Gy (range, 40 to 54 Gy). Fifty-nine patients also received fluoropyrimidine chemotherapy as a radiosensitizer. p-Akt, p-mTOR, and PTEN expression were assessed with immunohistochemical staining on the tissue microarray. p-Akt, p-mTOR, and PTEN were expressed in 23 (36.5%), 17 (27.0%), and 24 patients (38.1%), respectively. p-Akt expression was associated with distant metastasis and overall survival (OS), but not with locoregional recurrence. The 5-year distant metastasis-free and OS rates were 25.8% versus 58.2% (P=0.007), and 27.5% versus 50.2% (P=0.0167) in patients with negative and positive expression, respectively. On multivariate analysis, nodal involvement was the only significant prognosticator predicting inferior distant metastasis-free survival (P=0.0105), whereas p-Akt expression had a borderline significance (P=0.0541). As for OS, p-Akt expression was a marginally significant prognosticator (P=0.0635), whereas other risk factors lost the statistical significance. p-Akt expression tended to be associated with a favorable prognosis in patients undergoing curative resection followed by adjuvant CRT for proximal EHBD cancer.


A near-perfect invisibility cloak constructed with homogeneous materials.
A near-perfect, non-singular cylindrical invisibility cloak with diamond cross section is achieved by a two-step coordinate transformation. A small line segment is stretched and then blown up into a diamond space, and finally the cloak consisting of four kinds and eight blocks of homogeneous transformation media is obtained. Numerical simulations confirm the well performance of the cloak. The operation bandwidth of the cloak is also investigated. Our scheme is promising to create a simple and well-performed cloak in practice.


Widely tunable, narrow-linewidth, subnanosecond pulse generation in an electronically tuned Ti:sapphire laser.
Widely tunable subnanosecond Ti:sapphire laser radiation pumped with a cw Q -switched laser-diode-pumped Nd:YAG laser has been demonstrated in a simple laser system with a configuration of variable cavity length. Laser wavelengths can be continuously tuned by adjustment of the rf of an intracavity acousto-optic tunable filter with a computer through the whole range of the laser gain. During tuning of the whole spectral range, there is no need to realign any optics in the laser, except for moving the mirror to track the change of the rf. The peak powers of the output pulses at a pump level of 300 mW are comparable with those of conventional tunable picosecond Ti:sapphire lasers at a cw pump level of almost 10 W.


Autocatalysis: at the root of self-replication.
Autocatalysis is a fundamental concept, used in a wide range of domains. From its most general definition, that is, a process in which a chemical compound is able to catalyze its own formation, several different systems can be described. We detail the different categories of autocatalyses, and compare them on the basis of their mechanistic, kinetic, and dynamic properties. It is shown how autocatalytic patterns can be generated by different systems of chemical reactions. The notion of autocatalysis covers a large variety of mechanistic realizations with very similar behaviors; it is proposed that its key signature is its kinetic pattern expressed in a mathematical form. This notion, while describing dynamic behaviors at the most fundamental level, is at the basis for developing higher-level concepts towards life: autocatalytic sets, and autopoietic systems.


Analyzing Hospital Transfers Using INTERACT Acute Care Transfer Tools: Lessons from MOQI.
We explored the differences in potentially avoidable/unavoidable hospital transfers in a retrospective analysis of Interventions to Reduce Acute Care Transfers (INTERACT) Acute Transfer Tools (ACTs) completed by advanced practice registered nurses (APRNs) working in the Missouri Quality Improvement (QI) Initiative (MOQI). Cross-sectional descriptive study of 3996 ACTs for 32.5 calendar months from 2014 to 2016. Univariate analyses examined differences between potentially avoidable vs unavoidable transfers. Multivariate logistic regression analysis of candidate factors identified those contributing to avoidable transfers. Sixteen nursing homes (NHs), ranging from 120 to 321 beds, in urban, metro, and rural communities within 80 miles of a large midwestern city. A total of 5168 residents with a median age of 82 years. Data from 3946 MOQI-adapted ACTs. A total of 54% of hospital transfers were identified as avoidable. QI opportunities related to avoidable transfers were earlier detection of new signs/symptoms (odds ratio [OR] = 2.35; 95% confidence interval [CI] = 1.61-3.42; P < .001); discussions of resident/family preference (OR = 2.12; 95% CI = 1.38-3.25; P < .001); advance directive/hospice care (OR = 2.25; 95% CI = 1.33-3.82; P = .003); better communication about condition (OR = 4.93; 95% CI = 3.17-7.68; P < .001); and condition could have been managed in the NH (OR = 16.63; 95% CI = 10.9-25.37; P < .001). Three factors related to unavoidable transfers were bleeding (OR = .59; 95% CI = .46-.77; P < .001), nausea/vomiting (OR = .7; 95% CI = .54-.91; P = .007), and resident/family preference for hospitalization (OR = .79; 95% CI = .68-.93; P = .003). Reducing avoidable hospital transfers in NHs requires challenging assumptions about what is avoidable so QI efforts can be directed to improving NH capacity to manage ill residents. The APRNs served as the onsite coaches in the use and adoption of INTERACT. Changes in health policy would provide a revenue stream to support APRN presence in NH, a role that is critical to improving resident outcomes by increasing staff capacity to identify illness and guide system change. J Am Geriatr Soc 67:1953-1959, 2019.


Gradients of lipid storage, photosynthesis and plastid differentiation in developing soybean seeds.
This study establishes a topographical framework for functional investigations on the regulation of lipid biosynthesis and its interaction with embryo photosynthesis in developing soybean seed. Structural observations, combined with molecular and functional parameters, revealed the gradual transformation of chloroplasts into storage organelles, starting from inner regions going outwards. This is evidenced by electron microscopy, confocal laser scanning microscopy, in situ hybridization and histochemical/biochemical data. As a consequence of plastid differentiation, photosynthesis becomes distributed along a gradient within the developing embryo. Electron transport rate, effective quantum yield and O2 production rate are maximal in the embryo periphery, as documented by imaging pulse-amplitude-modulated fluorescence and O2 release via microsensors. The gradual loss of photosynthetic capacity was accompanied by a similarly gradual accumulation of starch and lipids. Noninvasive nuclear magnetic resonance spectroscopy of mature seeds revealed steep gradients in lipid deposition, with the highest concentrations in inner regions. The inverse relationship between photosynthesis and lipid biosynthesis argues against a direct metabolic involvement of photosynthesis in lipid biosynthesis during the late storage stage, but points to a role for photosynthetic oxygen release. This hypothesis is verified in a companion paper.


Natural extracellular matrix scaffolds recycled from human salivary digests: a morphometric study.
A challenge in engineering tissues is to supply parenchymal cells with suitable scaffolds which ideally reproduce the extracellular matrix (ECM). This study tested the hypothesis of preserving the 'residual connective tissue' remaining after mechanical and enzymatic release of cells from human submandibular gland biopsies (that we named 'natural ExtraCellular Matrix scaffolds', nECMsc) to be used as recycled natural scaffolds. The objective was to test whether nECMsc and native salivary tissue were comparable morphologically, in ECM proteins composition, and in cell seeding efficiency. Following cell isolation procedures, nECMsc were kept, either fresh or frozen (sectioned into 12-μm-thick slices), and examined with high-resolution electron microscopy (HRSEM) for its three-dimensional structure, and with picrosirius red staining and immunogold staining for ECM protein composition and distribution, respectively. nECMsc were seeded with human epithelial cells and fibroblasts to assess cell attachment and proliferation in short-term experiments. Under HRSEM, nECMsc had comparable fiber arrangement to original glands. Histochemical and immunogold-labeling examinations revealed the presence of collagen types I, III, and IV. Seeded epithelial cells and fibroblasts attached, proliferated (14-55%), and were alive (86-99%) after 4-8 days of culture. nECMsc retained native ECM proteins and maintained their distribution. Seeded cells remained viable on nECMsc.


Glycofullerenes Inhibit Particulate Matter Induced Inflammation and Loss of Barrier Proteins in HaCaT Human Keratinocytes.
Exposure to particulate matter (PM) has been linked to pulmonary and cardiovascular dysfunctions, as well as skin diseases, etc. PM impairs the skin barrier functions and is also involved in the initiation or exacerbation of skin inflammation, which is linked to the activation of reactive oxygen species (ROS) pathways. Fullerene is a single C60 molecule which has been reported to act as a good radical scavenger. However, its poor water solubility limits its biological applications. The glyco-modification of fullerenes increases their water solubility and anti-bacterial and anti-virus functions. However, it is still unclear whether it affects their anti-inflammatory function against PM-induced skin diseases. Hence, glycofullerenes were synthesized to investigate their effects on PM-exposed HaCaT human keratinocytes. Our results showed that glycofullerenes could reduce the rate of PM-induced apoptosis and ROS production, as well as decrease the expression of downstream mitogen-activated protein kinase and Akt pathways. Moreover, PM-induced increases in inflammatory-related signals, such as cyclooxygenase-2, heme oxygenase-1, and prostaglandin E2, were also suppressed by glycofullerenes. Notably, our results suggested that PM-induced impairment of skin barrier proteins, such as filaggrin, involucrin, repetin, and loricrin, could be reduced by pre-treatment with glycofullerenes. The results of this study indicate that glycofullerenes could be potential candidates for treatments against PM-induced skin diseases and that they exert their protective effects via ROS scavenging, anti-inflammation, and maintenance of the expression of barrier proteins.


Neo-adjuvant chemotherapy alone or with regional hyperthermia for localised high-risk soft-tissue sarcoma: a randomised phase 3 multicentre study.
The optimum treatment for high-risk soft-tissue sarcoma (STS) in adults is unclear. Regional hyperthermia concentrates the action of chemotherapy within the heated tumour region. Phase 2 studies have shown that chemotherapy with regional hyperthermia improves local control compared with chemotherapy alone. We designed a parallel-group randomised controlled trial to assess the safety and efficacy of regional hyperthermia with chemotherapy. Patients were recruited to the trial between July 21, 1997, and November 30, 2006, at nine centres in Europe and North America. Patients with localised high-risk STS (> or = 5 cm, Fédération Nationale des Centres de Lutte Contre le Cancer [FNCLCC] grade 2 or 3, deep to the fascia) were randomly assigned to receive either neo-adjuvant chemotherapy consisting of etoposide, ifosfamide, and doxorubicin (EIA) alone, or combined with regional hyperthermia (EIA plus regional hyperthermia) in addition to local therapy. Local progression-free survival (LPFS) was the primary endpoint. Efficacy analyses were done by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT 00003052. 341 patients were enrolled, with 169 randomly assigned to EIA plus regional hyperthermia and 172 to EIA alone. All patients were included in the analysis of the primary endpoint, and 332 patients who received at least one cycle of chemotherapy were included in the safety analysis. After a median follow-up of 34 months (IQR 20-67), 132 patients had local progression (56 EIA plus regional hyperthermia vs 76 EIA). Patients were more likely to experience local progression or death in the EIA-alone group compared with the EIA plus regional hyperthermia group (relative hazard [RH] 0.58, 95% CI 0.41-0.83; p=0.003), with an absolute difference in LPFS at 2 years of 15% (95% CI 6-26; 76% EIA plus regional hyperthermia vs 61% EIA). For disease-free survival the relative hazard was 0.70 (95% CI 0.54-0.92, p=0.011) for EIA plus regional hyperthermia compared with EIA alone. The treatment response rate in the group that received regional hyperthermia was 28.8%, compared with 12.7% in the group who received chemotherapy alone (p=0.002). In a pre-specified per-protocol analysis of patients who completed EIA plus regional hyperthermia induction therapy compared with those who completed EIA alone, overall survival was better in the combined therapy group (HR 0.66, 95% CI 0.45-0.98, p=0.038). Leucopenia (grade 3 or 4) was more frequent in the EIA plus regional hyperthermia group compared with the EIA-alone group (128 of 165 vs 106 of 167, p=0.005). Hyperthermia-related adverse events were pain, bolus pressure, and skin burn, which were mild to moderate in 66 (40.5%), 43 (26.4%), and 29 patients (17.8%), and severe in seven (4.3%), eight (4.9%), and one patient (0.6%), respectively. Two deaths were attributable to treatment in the combined treatment group, and one death was attributable to treatment in the EIA-alone group. To our knowledge, this is the first randomised phase 3 trial to show that regional hyperthermia increases the benefit of chemotherapy. Adding regional hyperthermia to chemotherapy is a new effective treatment strategy for patients with high-risk STS, including STS with an abdominal or retroperitoneal location. Deutsche Krebshilfe, Helmholtz Association (HGF), European Organisation of Research and Treatment of Cancer (EORTC), European Society for Hyperthermic Oncology (ESHO), and US National Institute of Health (NIH).


Feasibility and acceptability of a Takeaway Masterclass aimed at encouraging healthier cooking practices and menu options in takeaway food outlets.
To evaluate the feasibility and acceptability of the Takeaway Masterclass, a three-hour training session delivered to staff of independent takeaway food outlets that promoted healthy cooking practices and menu options. A mixed-methods study design. All participating food outlets provided progress feedback at 6 weeks post-intervention. Baseline and 6-week post-intervention observational and self-reported data were collected in half of participating takeaway food outlets. North East England. Independent takeaway food outlet owners and managers. Staff from eighteen (10 % of invited) takeaway food outlets attended the training; attendance did not appear to be associated with the level of deprivation of food outlet location. Changes made by staff that required minimal effort or cost to the business were the most likely to be implemented and sustained. Less popular changes included using products that are difficult (or expensive) to source from suppliers, or changes perceived to be unpopular with customers. The Takeaway Masterclass appears to be a feasible and acceptable intervention for improving cooking practices and menu options in takeaway food outlets for those who attended the training. Further work is required to increase participation and retention and explore effectiveness, paying particular attention to minimising adverse inequality effects.


Helping hands, healthy body? Oxytocin receptor gene and prosocial behavior interact to buffer the association between stress and physical health.
Providing help or support to others buffers the associations between stress and physical health. We examined the function of the neurohormone oxytocin as a biological mechanism for this stress-buffering phenomenon. Participants in a longitudinal study completed a measure of charitable behavior, and over the next two years provided assessments of stressful life events and physician-diagnosed physical ailments. Results indicated that charitable behavior buffered the associations between stressful events and new-onset ailments among individuals with the AA/AG genotypes of oxytocin receptor gene (OXTR) variant rs53576, but not among those with the GG genotype. These results suggest that oxytocin function may significantly affect health and may help explain the associations between prosocial behavior and health. More broadly, these findings are consistent with a role for the caregiving behavioral system in health and well-being.


Fatal Lyme carditis and endodermal heterotopia of the atrioventricular node.
A fatal case of Lyme carditis occurring in a Suffolk farmworker is reported. Post-mortem examination of the heart showed pericarditis, focal myocarditis and prominent endocardial and interstitial fibrosis. The additional finding of endodermal heterotopia ('mesothelioma') of the atrioventricular node raises the possibility that this could also be related to Lyme infection and account for the relatively frequent occurrence of atrioventricular block in this condition. Lyme disease should always be considered in a case of atrioventricular block, particularly in a young patient from a rural area. The heart block tends to improve and therefore only temporary pacing may be required.


Characterization of late response genes sequentially expressed during renewed growth of fibroblastic cells.
Many proto-oncogenes are rapidly and transiently activated during the early stages of the cellular transition from a resting G0 state to the DNA synthesis (S) phase. To get better understanding of the gene complexity involved at later stages, we isolated, by cDNA cloning, and identified 17 genes that are activated sequentially during the period of time from proto-oncogene expression to the onset of DNA synthesis in the hamster CCL39 fibroblastic cell line. When protein synthesis is inhibited, induced expression of these genes is unaffected for 10 of them, enhanced for four, in a fashion similar to the immediate-early response genes, and inhibited for three, as observed for delayed early-response genes. In addition to rhoG, a new member of the ras homolog gene family (Vincent et al., 1992), cDNA sequencing indicated that six of them correspond to cytoskeletal proteins (alpha-tubulin, vascular alpha-actin and skeletal gamma-actin), extracellular matrix protein (thrombospondin), secreted protease (plasminogen activator inhibitor-1) and energy-linked transporter (mitochondrial proton/phosphate symporter). This overall survey shows that numerous differentially regulated gene activations are associated with the cell cycle progression, and suggests that proteins involved in cellular reshaping participate actively in the control of cellular growth.


Amyotrophic lateral sclerosis caregiver burden and patients' quality of life during COVID-19 pandemic.
Objective: To assess patients Quality of life (QoL) and the burden of their caregivers during Covid-19 pandemic and specifically the impact of two-month lockdown period. Methods: In April 2020, a total of 60 patients and 59 caregivers were administered by phone scales assessing patients' QoL (McGill QoL Questionnaire), general health status (EQ-5D-5L), and caregiver burden (Zarit Burden Interview). The administration was repeated one month after the end of lockdown measures, with the addition of a qualitative questionnaire (COVID-QoL Questionnaire) exploring family reorganization and personal perception of lock down. Results: QoL and perceived health status did not worsen during lockdown, while caregiver burden increased (p = 0.01). Patient's QoL and caregiver burden were inversely correlated at T1 (ZBI total score mildly correlated with Mc Gill existential subscore, p = 0.02, rho = 0.30 and with Mc Gill total score, p = 0.05, rho = 0.265). No significant correlations were found at T2. According to the COVID-QoL questionnaire, caregivers perceived lower family help compared to patients (p < 0.001). Conclusions: Restricted measures of lockdown period during COVID-19 pandemic did not result in a significant reduction of QoL in our cohort of ALS patients, while caregiver burden significantly increased. ALS motor impairment may have played a role in the unchanged life conditions of patients. Instead, the restriction of family help for primary caregivers could be responsible of their increased burden, reflecting the importance of a wide social support in the management of this clinical condition.


Optimization of the dorsal skinfold window chamber model and multi-parametric characterization of tumor-associated vasculature.
The dorsal skinfold window chamber (DSWC) model is a unique tool that enables analysis of various aspects of tumor biology and therapeutic response. Although the protocol for the murine DSWC model is standardized, certain tumors fail to grow or require a particular environment to promote growth. Given such limitations, we optimized the DSWC model for a slow-growing tumor that regresses spontaneously in the standard protocol. We further characterized the vascular network in the tumor model compared with that of non-tumor-bearing mice and observed significant differences in multiple parameters related to vascular structure and function.


Action plans with brief patient education for exacerbations in chronic obstructive pulmonary disease.
Exacerbations of chronic obstructive pulmonary disease (COPD) are a major driver of decline in health status and impose high costs on healthcare systems. Action plans offer a form of self-management that can be delivered in the outpatient setting to help individuals recognise and initiate early treatment for exacerbations, thereby reducing their impact. To compare effects of an action plan for COPD exacerbations provided with a single short patient education component and without a comprehensive self-management programme versus usual care. Primary outcomes were healthcare utilisation, mortality and medication use. Secondary outcomes were health-related quality of life, psychological morbidity, lung function and cost-effectiveness. We searched the Cochrane Airways Group Specialised Register along with CENTRAL, MEDLINE, Embase and clinical trials registers. Searches are current to November 2015. We handsearched bibliographic lists and contacted study authors to identify additional studies. We included randomised controlled trials (RCT) and quasi-RCTs comparing use of an action plan versus usual care for patients with a clinical diagnosis of COPD. We permitted inclusion of a single short education component that would allow individualisation of action plans according to management needs and symptoms of people with COPD, as well as ongoing support directed at use of the action plan. We used standard methodological procedures expected by Cochrane. For meta-analyses, we subgrouped studies via phone call follow-up directed at facilitating use of the action plan. This updated review includes two additional studies (and 976 additional participants), for a total of seven parallel-group RCTs and 1550 participants, 66% of whom were male. Participants' mean age was 68 years and was similar among studies. Airflow obstruction was moderately severe in three studies and severe in four studies; mean post bronchodilator forced expiratory volume in one second (FEV1) was 54% predicted, and 27% of participants were current smokers. Four studies prepared individualised action plans, one study an oral plan and two studies standard written action plans. All studies provided short educational input on COPD, and two studies supplied ongoing support for action plan use. Follow-up was 12 months in four studies and six months in three studies.When compared with usual care, an action plan with phone call follow-up significantly reduced the combined rate of hospitalisations and emergency department (ED) visits for COPD over 12 months in one study with 743 participants (rate ratio (RR) 0.59, 95% confidence interval (CI) 0.44 to 0.79; high-quality evidence), but the rate of hospitalisations alone in this study failed to achieve statistical significance (RR 0.69, 95% CI 0.47 to 1.01; moderate-quality evidence). Over 12 months, action plans significantly decreased the likelihood of hospital admission (odds ratio (OR) 0.69, 95% CI 0.49 to 0.97; n = 897; two RCTs; moderate-quality evidence; number needed to treat for an additional beneficial outcome (NNTB) 19 (11 to 201)) and the likelihood of an ED visit (OR 0.55, 95% CI 0.38 to 0.78; n = 897; two RCTs; moderate-quality evidence; NNTB over 12 months 12 (9 to 26)) compared with usual care.Results showed no significant difference in all-cause mortality during 12 months (OR 0.88, 95% CI 0.59 to 1.31; n = 1134; four RCTs; moderate-quality evidence due to wide confidence interval). Over 12 months, use of oral corticosteroids was increased with action plans compared with usual care (mean difference (MD) 0.74 courses, 95% CI 0.12 to 1.35; n = 200; two RCTs; moderate-quality evidence), and the cumulative prednisolone dose was significantly higher (MD 779.0 mg, 95% CI 533.2 to 10248; n = 743; one RCT; high-quality evidence). Use of antibiotics was greater in the intervention group than in the usual care group (subgrouped by phone call follow-up) over 12 months (MD 2.3 courses, 95% CI 1.8 to 2.7; n = 943; three RCTs; moderate-quality evidence).Subgroup analysis by ongoing support for action plan use was limited; review authors noted no subgroup differences in the likelihood of hospital admission or ED visits or all-cause mortality over 12 months. Antibiotic use over 12 months showed a significant difference between subgroups in studies without and with ongoing support.Overall quality of life score on St George's Respiratory Questionnaire (SGRQ) showed a small improvement with action plans compared with usual care over 12 months (MD -2.8, 95% CI -0.8 to -4.8; n = 1009; three RCTs; moderate-quality evidence). Low-quality evidence showed no benefit for psychological morbidity as measured with the Hospital Anxiety and Depression Scale (HADS). Use of COPD exacerbation action plans with a single short educational component along with ongoing support directed at use of the action plan, but without a comprehensive self-management programme, reduces in-hospital healthcare utilisation and increases treatment of COPD exacerbations with corticosteroids and antibiotics. Use of COPD action plans in this context is unlikely to increase or decrease mortality. Whether additional benefit is derived from periodic ongoing support directed at use of an action plan cannot be determined from the results of this review.


Speciation despite globally overlapping distributions in Penicillium chrysogenum: the population genetics of Alexander Fleming's lucky fungus.
Eighty years ago, Alexander Fleming described the antibiotic effects of a fungus that had contaminated his bacterial culture, kick starting the antimicrobial revolution. The fungus was later ascribed to a putatively globally distributed asexual species, Penicillium chrysogenum. Recently, the species has been shown to be genetically diverse, and possess mating-type genes. Here, phylogenetic and population genetic analyses show that this apparently ubiquitous fungus is actually composed of at least two genetically distinct species with only slight differences detected in physiology. We found each species in air and dust samples collected in and around St Mary's Hospital where Fleming worked. Genotyping of 30 markers across the genome showed that preserved fungal material from Fleming's laboratory was nearly identical to derived strains currently in culture collections and in the same distinct species as a wild progenitor strain of current penicillin producing industrial strains rather than the type species P. chrysogenum. Global samples of the two most common species were found to possess mating-type genes in a near 1:1 ratio, and show evidence of recombination with little geographic population subdivision evident. However, no hybridization was detected between the species despite an estimated time of divergence of less than 1MYA. Growth studies showed significant interspecific inhibition by P. chrysogenum of the other common species, suggesting that competition may facilitate species maintenance despite globally overlapping distributions. Results highlight under-recognized diversity even among the best-known fungal groups and the potential for speciation despite overlapping distribution.


A measure of perceived pain and tobacco smoking interrelations: pilot validation of the pain and smoking inventory.
Mounting evidence indicates that pain can motivate cigarette smoking behavior, smokers have reliably endorsed the use of tobacco to cope with pain, and there is reason to suspect that pain may impede smoking cessation. Smoking-related outcome expectancies are among the best predictors of cigarette consumption and relapse, and the goal of this pilot study was to develop a standardized measure of how tobacco smokers perceive their pain and smoking behavior to be interrelated (i.e. pain as a motivator of smoking, expectancies for smoking as a means of coping with pain, and pain as a barrier to quitting). Participants (N = 75) completed an online survey that was designed to assess interrelations between pain and tobacco smoking. The nine-item Pain and Smoking Inventory (PSI) demonstrated excellent internal consistency (α = .95). As expected, PSI scores were higher among smokers with chronic pain (vs. no pain), and were positively associated with established indices of both pain and tobacco smoking dependence. These results corroborate the notion that smokers can effectively communicate perceptions of interrelations between their pain and smoking behavior. Future research is needed to establish reliability and validity of the PSI among larger, more diverse samples of smokers with varying levels of pain. Future work should also examine PSI scores as predictors of smoking cessation outcomes, and whether PSI data may usefully inform the development of tailored interventions for smokers in pain.


Assessing the essentiality of the decaprenyl-phospho-d-arabinofuranose pathway in Mycobacterium tuberculosis using conditional mutants.
In Mycobacterium tuberculosis the decaprenyl-phospho-d-arabinofuranose (DPA) pathway is a validated target for the drugs ethambutol and benzothiazinones. To identify other potential drug targets in the pathway, we generated conditional knock-down mutants of each gene involved using the TET-PIP OFF system. dprE1, dprE2, ubiA, prsA, rv2361c, tkt and rpiB were confirmed to be essential under non-permissive conditions, whereas rv3807c was not required for survival. In the most vulnerable group, DprE1-depleted cells died faster in vitro and intracellularly than those lacking UbiA and PrsA. Downregulation of DprE1 and UbiA resulted in similar phenotypes, namely swelling of the bacteria, cell wall damage and lysis as observed at the single cell level, by real time microscopy and electron microscopy. By contrast, depletion of PrsA led to cell elongation and implosion, which was suggestive of a more pleiotropic effect. Drug sensitivity assays with known DPA-inhibitors supported the use of conditional knock-down strains for target-based whole-cell screens. Together, our work provides strong evidence for the vulnerability of all but one of the enzymes in the DPA pathway and generates valuable tools for the identification of lead compounds targeting the different biosynthetic steps. PrsA, phosphoribosyl-pyrophosphate synthetase, appears to be a particularly attractive new target for drug discovery.


Perspectives from the Avian Phylogenomics Project: Questions that Can Be Answered with Sequencing All Genomes of a Vertebrate Class.
The rapid pace of advances in genome technology, with concomitant reductions in cost, makes it feasible that one day in our lifetime we will have available extant genomes of entire classes of species, including vertebrates. I recently helped cocoordinate the large-scale Avian Phylogenomics Project, which collected and sequenced genomes of 48 bird species representing most currently classified orders to address a range of questions in phylogenomics and comparative genomics. The consortium was able to answer questions not previously possible with just a few genomes. This success spurred on the creation of a project to sequence the genomes of at least one individual of all extant ∼10,500 bird species. The initiation of this project has led us to consider what questions now impossible to answer could be answered with all genomes, and could drive new questions now unimaginable. These include the generation of a highly resolved family tree of extant species, genome-wide association studies across species to identify genetic substrates of many complex traits, redefinition of species and the species concept, reconstruction of the genomes of common ancestors, and generation of new computational tools to address these questions. Here I present visions for the future by posing and answering questions regarding what scientists could potentially do with available genomes of an entire vertebrate class.


A case of Buerger's disease.
Buerger's disease (thromboangiitis obliterans) is an uncommon vascular occlusive disease most commonly affecting the lower extremities of young male tobacco smokers. We report the case of a thirty-five-year-old man who presented with nonhealing ulcerations of several toes. Arteriography revealed severe bilateral vascular occlusion. The patient failed to respond to intensive topical care, nifedipine, pentoxifylline, hyperbaric oxygen, intravenous antibiotics, and whirlpool. He continued to smoke, and eventually became septic, requiring amputations of his gangrenous left second toe and right leg below the knee.


A review of the impact and effectiveness of nurse-led care in dermatology.
To identify systematically, summarize and critically appraise the current evidence regarding the impact and effectiveness of nurse-led care in dermatology. A diverse range of nurse-led models of care exist in dermatology. Primary studies have been conducted evaluating these models, but review and synthesis of the findings from these studies have not been undertaken. Systematic searches of CINAHL, MEDLINE, British Nursing Index (BNI) and the RCN Library Catalogue from 1990 until March 2005. The searches were supplemented by an extensive hand search of the literature through references identified from retrieved articles and by contact with experts in the field. Fourteen relevant publications were identified and included findings from both primary and secondary care. The evidence indicates that nurses are treating a number of dermatological conditions, primarily using treatment protocols, across a broad range of clinical settings. However, some nurses working in primary care, lack confidence to treat some of these conditions and the educational needs of these nurses are frequently unmet. A reduction in the severity of the condition and more effective use of topical therapies are benefits of nurse interventions on service delivery. Faster access to treatment, a reduction in referrals to the general practitioner or dermatologist and an increase in knowledge of their condition are benefits reported by patients. Findings of the review are generally positive. However, there are methodological weaknesses and under researched issues, e.g. cost effectiveness of nurse-led care and the prescription of medicines by nurses for patients with dermatological conditions that point to the need for further rigorous evaluation. Nurse-led care is an integral element of the dermatology service offered to patients. This review highlights the impact of this care and the issues that require consideration by those responsible for the development of nurse-led models of care in dermatology.


Optical processing between two metallically hybrid microdisks.
A waveguide is bridged between two metallically hybrid microdisks for achieving nonlocal optical processing. The whispering gallery mode (WGM) is split into two modes, one is intrinsic WGM and the other is oscillating mode (OM) whose photons oscillate between two microdisks through the waveguide. The characteristics of OM on the waveguide's width and length are studied. The results show the fundamental WGM oscillation only happens at certain lengths and widths of the waveguide, and strong optical interaction between two microdisks is accompanied by a high-loss waveguide. The optical processing through a waveguide between two individual resonators or photon producers may be applied to dynamic control on the photon states.


Caring for a family member with intellectual disability and epilepsy: practical, social and emotional perspectives.
To examine the caregiving impact of those who support a family member with intellectual disability and epilepsy. An online, qualitative international survey was conducted via the auspices of the International Bureau of Epilepsy with various stakeholders who support individuals who have intellectual disability and epilepsy. Qualitative comments were analyzed from respondents who identified themselves as family members (n=48; 36%) who referred specifically to the impact of supporting a family member with these combined disabilities. Four main domains, which were comprised of ten themes, were derived from the qualitative data using Braun and Clarke's qualitative framework. These domains comprised (1) practical concerns, (2) disrupted family dynamics, (3) emotional burden and (4) positive experiences. In combination these themes illustrate the pervasive impact on family life for those supporting an individual with complex needs. Financial concerns, coordination and responsibility of care, diverted attention from other family members and social isolation all contributed a significant burden of care for family members. Positive aspects were, however, also cited including the closeness of the family unit and a fostering of altruistic behavior. The study provides an insight into an under-researched area. The burden of caring for a family member across the lifespan has a largely negative and pervasive impact. Targeted service provision could contribute to an amelioration of the challenges faced by these families.


Thermal and laser properties of Yb:LuAG for kW thin disk lasers.
Thin disk laser experiments with Yb:LuAG (Yb:Lu(3)Al(5)O(12)) were performed leading to 5 kW of output power and an optical-to-optical efficiency exceeding 60%. Comparative analyses of the laser relevant parameters of Yb:LuAG and Yb:YAG were carried out. While the spectroscopic properties were found to be nearly identical, investigations of the thermal conductivities revealed a 20% higher value for Yb:LuAG at Yb(3+)-doping concentrations of about 10%. Due to the superior thermal conductivity with respect to Yb:YAG, Yb:LuAG offers thus the potential of improved performance in high power thin disk laser applications.


Rivaroxaban versus enoxaparin for thromboprophylaxis after total knee arthroplasty.
We investigated the efficacy of rivaroxaban, an orally active direct factor Xa inhibitor, in preventing venous thrombosis after total knee arthroplasty. In this randomized, double-blind trial, 2531 patients who were to undergo total knee arthroplasty received either oral rivaroxaban, 10 mg once daily, beginning 6 to 8 hours after surgery, or subcutaneous enoxaparin, 40 mg once daily, beginning 12 hours before surgery. The primary efficacy outcome was the composite of any deep-vein thrombosis, nonfatal pulmonary embolism, or death from any cause within 13 to 17 days after surgery. Secondary efficacy outcomes included major venous thromboembolism (i.e., proximal deep-vein thrombosis, nonfatal pulmonary embolism, or death related to venous thromboembolism) and symptomatic venous thromboembolism. The primary safety outcome was major bleeding. The primary efficacy outcome occurred in 79 of 824 patients (9.6%) who received rivaroxaban and in 166 of 878 (18.9%) who received enoxaparin (absolute risk reduction, 9.2%; 95% confidence interval [CI], 5.9 to 12.4; P<0.001). Major venous thromboembolism occurred in 9 of 908 patients (1.0%) given rivaroxaban and 24 of 925 (2.6%) given enoxaparin (absolute risk reduction, 1.6%; 95% CI, 0.4 to 2.8; P=0.01). Symptomatic events occurred less frequently with rivaroxaban than with enoxaparin (P=0.005). Major bleeding occurred in 0.6% of patients in the rivaroxaban group and 0.5% of patients in the enoxaparin group. The incidence of drug-related adverse events, mainly gastrointestinal, was 12.0% in the rivaroxaban group and 13.0% in the enoxaparin group. Rivaroxaban was superior to enoxaparin for thromboprophylaxis after total knee arthroplasty, with similar rates of bleeding. (ClinicalTrials.gov number, NCT00361894.)


Evidence-based guidelines for treating bipolar disorder: recommendations from the British Association for Psychopharmacology.
The British Association for Psychopharmacology guidelines specify the scope and target of treatment for bipolar disorder. They are based explicitly on the available evidence and presented, similar to previous Clinical Practice guidelines, as recommendations to aid clinical decision-making for practitioners. They may also serve as a source of information for patients and carers. The recommendations are presented together with a more detailed review of the available evidence. A consensus meeting, involving experts in bipolar disorder and its treatment, reviewed key areas and considered the strength of evidence and clinical implications. The guidelines were drawn up after extensive feedback from participants and interested parties. The strength of supporting evidence was rated. The guidelines cover the diagnosis of bipolar disorder, clinical management and strategies for the use of medicines in short-term treatment of episodes, relapse prevention and stopping treatment.


Personality trends in keratoconus. An analysis.
Patients with keratoconus frequently are described as having peculiar personality characteristics, despite the lack of controlled studies in the ophthalmologic or psychiatric literature. We studied 109 subjects, using a standardized personality inventory (the Millon Clinical Multiaxial Inventory) that measures 20 personality scales-both normal and pathologic. Subjects were divided into three age-matched groups: (1) patients with keratoconus, (2) patients with other chronic eye diseases, and (3) normal controls. Results indicated that although chronic eye disease, including keratoconus, did have an impact on personality functioning in young and middle-aged adults, no specific complex of personality characteristics attributable to keratoconus could be identified. Patients with keratoconus differed from normal controls in much the same way as did patients with other chronic eye diseases, being less conforming and more passive-aggressive, paranoid, and hypomanic. They tended to more disorganized patterns of thinking and scored higher on substance abuse indicators. The influence of keratoconus on personality may be a function of the timing and nature of its onset in the context of the patient's psychosocial development.


The X-linked lymphoproliferative disease gene product SAP associates with PAK-interacting exchange factor and participates in T cell activation.
SLAM (signaling lymphocyte activation molecule)-associated protein (SAP) is a Src homology 2 (SH2) domain-containing adaptor expressed in T cells and natural killer cells. Its essential role in immune responses is underscored by the recent finding that mutations in SAP result in a rare but fatal X-linked lymphoproliferative disease (XLP). Although SAP is known to associate with SLAM-family receptors, the exact molecular mechanism by which SAP regulates lymphocyte signaling remains elusive. We here report that in T cells, SAP associates with the PAK-interacting exchange factor (PIX), a guanine nucleotide exchange factor (GEF) specific for Rac/Cdc42 GTPases. Moreover, SAP, PIX, and an activated form of Cdc42 form a complex in mammalian cells. We demonstrate that the SAP-PIX interaction is specific and is mediated by the C-terminal region of the SAP SH2 domain and the PIX SH3 domain. We further show that SAP is required for the recruitment of PIX to the SLAM-family receptors. Interestingly, overexpression of SAP, but not its homolog EAT-2, leads to a synergistic activation of nuclear factor of activating T cells (NFAT) in combination with a calcium signal in T cells. This SAP-mediated activation appears to be receptor-dependent and can be blocked by a dominant negative form of PIX. Taken together, our data strongly suggest that, in addition to the known SAP-interacting kinase Fyn, PIX may be another key player in SAP-mediated T cell activation.


A modification of the Bohr method to determine airways deadspace for non-uniform inspired gas tensions.
The Bohr method is a technique to determine airways deadspace using a tracer gas such as carbon dioxide or nitrogen. It is based on the assumption that the inspired concentration of the tracer gas is constant throughout inspiration. However, in some lung function measurement techniques where inspired concentration of the tracer gas may be required to vary, or where rapid injection of the tracer gas is made in real time, uniform inspired concentration is difficult or impossible to achieve, which leads to inaccurate estimation of deadspace using the Bohr equation. One such lung function measurement technique is the inspired sinewave technique. In this paper, we proposed a modification of the Bohr method, relaxing the requirement of absolute uniformity of tracer concentration in the inspired breath. The new method used integration of flow and concentration. A computer algorithm sought an appropriate value of deadspace to satisfy the mass balance equation for each breath. A modern gas mixing apparatus with rapid mass flow controllers was used to verify the procedure. Experiments on a tidally ventilated bench lung showed that the new method estimated dead space within 10% of the actual values whereas the traditional Bohr deadspace gave more than 50% error. The new method improved the accuracy of deadspace estimation when the inspired concentration is not uniform. This improvement would lead to more accurate diagnosis and more accurate estimations of other lung parameters such as functional residual capacity and pulmonary blood flow.


Blocking of feline immunodeficiency virus infection by a monoclonal antibody to CD9 is via inhibition of virus release rather than interference with receptor binding.
A monoclonal antibody, MAb vpg15, inhibits feline immunodeficiency virus (FIV) infection in tissue culture. The antibody is directed to a determinant of the feline cell surface marker, CD9, implying that CD9 may serve as a viral receptor or coreceptor in this system. In cells expressing CD9, MAb vpg15 markedly delayed acute virus infection in terms of reverse transcriptase activity detected in cell culture supernatants. This effect was evident if the antibody was added before, immediately after, or 24 h after virus infection. Binding experiments showed that MAb vpg15 did not block virus binding to the cells. PCR analyses at various intervals postinfection also indicated that MAb vpg15 did not block virus uptake, reverse transcription of viral RNA, or integration into host cell DNA. Multiply spliced mRNAs were detected up to 24 h postinfection in both control and MAb vpg15-treated cells. However, viral mRNAs were markedly diminished in MAb vpg15-treated cells after this time, consistent with a failure of the FIV infection to spread in the cell culture. Treatment of chronically infected cells with MAb vpg15 also caused a sharp diminution in viral particle production, while viral mRNA levels were the same in both untreated and MAb-treated infected cells. Analyses of intracellular and extracellular levels of virus-associated antigens showed an enhanced accumulation of intracellular p24. These findings are consistent with the interpretation that MAb vpg15 acts at a posttranscriptional stage by interfering with the assembly and/or release of virus from the cell.


The arbovirus vector Culex torrentium is more prevalent than Culex pipiens in northern and central Europe.
Two species of arbovirus vector, Culex torrentium and Culex pipiens (Diptera: Culicidae), occur in several European countries, but difficulties in their accurate identification and discrimination have hampered both detailed and large-scale distribution and abundance studies. Using a molecular identification method, we identified to species 2559 larvae of Cx. pipiens/torrentium collected from 138 sites in 13 European countries ranging from Scandinavia to the Mediterranean coast. In addition, samples of 1712 males of Cx. pipiens/torrentium collected at several sites in the Czech Republic were identified to species based on the morphology of their hypopygia. We found that the two species occur together in large areas of Europe, and that Cx. torrentium dominates in northern Europe and Cx. pipiens dominates south of the Alps. The transition in dominance occurs in central Europe, where both species are roughly equally common. There was a strong correlation between the length of the growing season at different sites and occurrences of the two species. As the growing season increases, the proportion and detection of Cx. torrentium decrease, whereas those of Cx. pipiens increase. The present findings have important consequences for the interpretation of the results of studies on major enzootic and link-vectors of mosquito-borne bird-associated viruses (i.e. Sindbis, West Nile and Usutu viruses), especially in central Europe and Scandinavia.


Bronchioloalveolar carcinoma: a model for investigating the biology of epidermal growth factor receptor inhibition.
Bronchioloalveolar carcinoma (BAC) is a previously uncommon subset of non-small cell lung cancer (NSCLC) with unique epidemiology, pathology, clinical features, radiographic presentation, and natural history compared with other NSCLC subtypes. Recent data suggest that the incidence of BAC is increasing, notably in younger nonsmoking women. Despite reports of prolonged survival after repeated surgical resection of multifocal lesions and slow growth kinetics, advanced bilateral or recurrent diffuse BAC remains incurable, with the vast majority of patients dying of respiratory failure or intercurrent pneumonia within 5 years. Limited data suggest that chemotherapy may yield poor results in BAC. However, anecdotal reports of prolonged complete response to tyrosine kinase inhibitors of the epidermal growth factor receptor (EGFR), a member of the human epidermal growth factor receptor (erbB) family, have raised considerable interest in studying this NSCLC subset. Here we present clinical data and preliminary results of correlative science studies analyzing human epidermal growth factor receptor pathways from the following two prospective Southwest Oncology Group clinical trials performed in advanced stage BAC: S9714 testing a 96-h continuous infusion of paclitaxel (Taxol) and S0126 evaluating the small molecule EGFR inhibitor gefitinib (ZD1839 or Iressa). These studies provide a biological rationale for investigating BAC as a model of predictive markers of EGFR inhibition.


Functional data analysis of single-trial auditory evoked potentials recorded in the awake rat.
Evoked potentials (EPs) reflect neural processing and are widely used to study sensory perception. However, methods of analyzing EP have been limited mostly to the conventional ensemble averaging of EP response trials to a repeated stimulus, and less so to single-trials analysis. Here we applied a new approach - functional data analysis (FDA) - to study auditory EP in the rat model of tinnitus, in which overdoses of salicylate (SS) are known to alter sound perception characteristically, as the same way as in humans. Single-trial auditory EPs were analyzed, after being collected on a daily basis from an awake rat, which had been surgically implanted with intracranial electrodes over its auditory cortex. Single-trial EP integrals were generated with sound stimuli (tones and clicks) presented systematically over an intensity range. The results were approximated using the cubic spline to give sets of smoothed response-level functions in dependence on the sound intensity. These functional data were analyzed using the methods of FDA. Comparisons between daily intensity series for each sound type were done using cross-distance measures based on the response-level functions in both the original form and the first-derivative form. From the results of FDA, the first-derivative form was found to provide a clearer separation when EP data from control groups were compared to the data from SS groups. This is also true when the daily data were compared within the more variable SS-group itself. In addition, at the high-intensity region where SS-action is presumably strong, we also observed characteristic changes in two statistical parameters, mean and skewness, of the cross-distance representations. Results suggested that FDA is a sensitive approach for EP studies, and it can become a powerful tool for the research in neural science, particularly neuropharmacology.


Psychological distress in Ghana: associations with employment and lost productivity.
Mental health disorders account for 13% of the global burden of disease, a burden that low-income countries are generally ill-equipped to handle. Research evaluating the association between mental health and employment in low-income countries, particularly in sub-Saharan Africa, is limited. We address this gap by examining the association between employment and psychological distress. We analyzed data from the Ghana Socioeconomic Panel Survey using logistic regression (N = 5,391 adults). In multivariable analysis, we estimated the association between employment status and psychological distress, adjusted for covariates. We calculated lost productivity from unemployment and from excess absence from work that respondents reported was because of their feelings of psychological distress. Approximately 21% of adults surveyed had moderate or severe psychological distress. Increased psychological distress was associated with increased odds of being unemployed. Men and women with moderate versus mild or no psychological distress had more than twice the odds of being unemployed. The association of severe versus mild or no distress with unemployment differed significantly by sex (P-value for interaction 0.004). Among men, the adjusted OR was 12.4 (95% CI: 7.2, 21.3), whereas the association was much smaller for women (adjusted OR = 3.8, 95% CI: 2.5, 6.0). Extrapolating these figures to the country, the lost productivity associated with moderate or severe distress translates to approximately 7% of the gross domestic product of Ghana. Psychological distress is strongly associated with unemployment in Ghana. The findings underscore the importance of addressing mental health issues, particularly in low-income countries.


Insomnia Really Hurts: Effect of a Bad Night's Sleep on Pain Increases With Insomnia Severity.
Insomnia and chronic pain are highly prevalent conditions and are often comorbid. Somatic complaints other than pain are also often observed in insomnia. Poor sleep and pain are known to mutually reinforce each other. However, it is unknown whether the habitual severity of insomnia modulates the acute effect of a particularly bad night's sleep on the next day's pain severity, and whether it modulates the acute effect of pain on the following night's sleep quality. Using data from 3,508 volunteers (2,684 female, mean age 50.09 y), we addressed these questions in addition to the associations between the habitual severity of insomnia, somatic complaints, and pain. Results indicated that people suffering from more severe habitual insomnia showed stronger mutual acute within-day reactivity of pain and poor sleep quality. The same increased reactivity was found in people with more severe habitual pain. Interestingly, the acute within-day mutual reactivity of pain and sleep quality showed consistent asymmetry. Pain worsened more after a particularly bad night's sleep than it improved after a particularly good night's sleep. Likewise, sleep worsened more after a day with more-than-usual pain than it improved after a day with less-than-usual pain. Future interventions may profit from addressing this asymmetric mutual reactivity especially in people with severe comorbid insomnia and chronic pain.


Post-meal perceivable satiety and subsequent energy intake with intake of partially hydrolysed guar gum.
Partially hydrolysed guar gum (PHGG), a soluble dietary fibre, has been shown to provide many health benefits. Previous studies had suggested that the combination of PHGG with protein provided a significant satiation effect on visual analogue scales (VAS). What was lacking was only the effect of administration of small doses of PHGG on post-meal satiation and subsequent energy intake. The objectives of the present investigations were to find the subjective perception of post-meal satiety with acute and long term administration of small amounts of PHGG alone with food, its effects on subsequent energy intake and the comparative effects among different types of soluble fibres. The following three separate studies were conducted: in study 1, healthy subjects (n 12) consumed PHGG along with breakfast, lunch and an evening snack; in study 2, healthy subjects (n 24) consumed 2 g of PHGG or dextrin along with yogurt as breakfast for 2 weeks; in study 3, healthy subjects (n 6) took 6 g each of either PHGG or indigestible dextrin or inulin along with lunch. In all the studies, various satiety parameters were measured on VAS before and after consumption of PHGG. The addition of PHGG showed significant (P < 0.05) acute (studies 1 and 3) and long-term (studies 1 and 2) satiety effects compared to the control and/or an equal amount of carbohydrate or other types of soluble fibre. Study 2 also indicated that the prolonged consumption of PHGG may significantly (P < 0.05) reduce energy intake from whole-day snacking. PHGG could be an ideal natural soluble fibre for delivering acute and long term satiety effects for comfortable appetite control.


Application of a Common Data Model (CDM) to rank the paediatric user and prescription prevalence of 15 different drug classes in South Korea, Hong Kong, Taiwan, Japan and Australia: an observational, descriptive study.
To measure the paediatric user and prescription prevalence in inpatient and ambulatory settings in South Korea, Hong Kong, Taiwan, Japan and Australia by age and gender. A further objective was to list the most commonly used drugs per drug class, per country. Hospital inpatient and insurance paediatric healthcare data from the following databases were used to conduct this descriptive drug utilisation study: (i) the South Korean Ajou University School of Medicine database; (ii) the Hong Kong Clinical Data Analysis and Reporting System; (iii) the Japan Medical Data Center; (iv) Taiwan's National Health Insurance Research Database and (v) the Australian Pharmaceutical Benefits Scheme. Country-specific data were transformed into the Observational Medical Outcomes Partnership Common Data Model. Children (≤18 years) with at least 1 day of observation in any of the respective databases from January 2009 until December 2013 were included. For each drug class, we assessed the per-protocol overall user and prescription prevalence rates (per 1000 persons) per country and setting. Our study population comprised 1 574 524 children (52.9% male). The highest proportion of dispensings was recorded in the youngest age category (<2 years) for inpatients (45.1%) with a relatively high user prevalence of analgesics and antibiotics. Adrenergics, antihistamines, mucolytics and corticosteroids were used in 10%-15% of patients. For ambulatory patients, the highest proportion of dispensings was recorded in the middle age category (2-11 years, 67.1%) with antibiotics the most dispensed drug overall. Country-specific paediatric drug utilisation patterns were described, ranked and compared between four East Asian countries and Australia. The widespread use of mucolytics in East Asia warrants further investigation.


Effects of living near an urban motorway on the wellbeing of local residents in deprived areas: Natural experimental study.
Health and wellbeing are partly shaped by the neighbourhood environment. In 2011, an eight kilometre (five mile) extension to the M74 motorway was opened in Glasgow, Scotland, constructed through a predominantly urban, deprived area. We evaluated the effects of the new motorway on wellbeing in local residents. This natural experimental study involved a longitudinal cohort (n = 365) and two cross-sectional samples (baseline n = 980; follow-up n = 978) recruited in 2005 and 2013. Adults from one of three study areas-surrounding the new motorway, another existing motorway, or no motorway-completed a postal survey. Within areas, individual measures of motorway proximity were calculated. Wellbeing was assessed with the mental (MCS-8) and physical (PCS-8) components of the SF-8 scale at both time points, and the short Warwick-Edinburgh Mental Well-being Scale (SWEMWBS) at follow-up only. In multivariable linear regression analyses, cohort participants living nearer to the new M74 motorway experienced significantly reduced mental wellbeing over time (MCS-8: -3.6, 95% CI -6.6 to -0.7) compared to those living further away. In cross-sectional and repeat cross-sectional analyses, an interaction was found whereby participants with a chronic condition living nearer to the established M8 motorway experienced reduced (MCS-8: -3.7, 95% CI -8.3 to 0.9) or poorer (SWEMWBS: -1.1, 95% CI -2.0 to -0.3) mental wellbeing compared to those living further away. We found some evidence that living near to a new motorway worsened local residents' wellbeing. In an area with an existing motorway, negative impacts appeared to be concentrated in those with chronic conditions, which may exacerbate existing health inequalities and contribute to poorer health outcomes. Health impacts of this type of urban regeneration intervention should be more fully taken into account in future policy and planning.


Apigenin inhibits in vitro and in vivo tumorigenesis in cisplatin-resistant colon cancer cells by inducing autophagy, programmed cell death and targeting m-TOR/PI3K/Akt signalling pathway.
Colon cancer is one of the deadly malignancies and the second most frequent cancer in the world. The development of drug resistance and dearth of the viable drug options forms an obstacle in the treatment of colon cancer. Herein, the anticancer potential of Apigenin was examined against cisplatin-resistant colon cancer cells. The proliferation rate of the cisplatin-resistant colon cancer cell line HT-29 was assessed by WST-1 assay. Autophagy was detected by electron microscopy. Apoptotic cell death was analysed by propidium iodide (PI) staining. Cell cycle analysis was performed by flow cytometry. Protein expression was determined by immuno blotting. Xenografted mice models were used for in vivo evaluation of Apigenin. The results showed that Apigenin could considerably inhibit the proliferation of colon cancer cells. The anticancer activity of Apigenin against the HT-29 colon cancer cells was found to be due to induction of autophagy and apoptosis. The Apigenin-triggered apoptosis and autophagy were also linked with alteration in the apoptosis and autophagy-related protein expression. Furthermore, it was found that Apigenin could inhibit the m-TOR/PI3K/AKT signalling pathway in the cisplatin-resistant colon cancer cells. The effects of Apigenin were also examined in vivo in xenografted mice models and it was revealed that Apigenin inhibited the growth of xenografted tumors. Taken together, these results indicate that Apigenin could inhibit the growth of cisplatin-resistant colon cancer cells in vitro and in vivo and may be used for the improvement of therapy of colon cancer.


Prospective changes in global DNA methylation and cancer incidence and mortality.
Methylation of repetitive elements Alu and LINE-1 in humans is considered a surrogate for global DNA methylation. Previous studies of blood-measured Alu/LINE-1 and cancer risk are inconsistent. We studied 1259 prospective methylation measurements from blood drawn 1-4 times from 583 participants from 1999 to 2012. We used Cox regression to evaluate time-dependent methylation as a biomarker for cancer risk and mortality, and linear regression to compare mean differences in methylation over time by cancer status and analyse associations between rate of methylation change and cancer. Time-dependent LINE-1 methylation was associated with prostate cancer incidence (HR: 1.38, 95% CI: 1.01-1.88) and all-cancer mortality (HR: 1.41, 95% CI: 1.03-1.92). The first measurement of Alu methylation (HR: 1.39, 95% CI: 1.08-1.79) was associated with all-cancer mortality. Participants who ultimately developed cancer had lower mean LINE-1 methylation than cancer-free participants 10+ years pre-diagnosis (P<0.01). Rate of Alu methylation change was associated with all-cancer incidence (HR: 3.62, 95% CI: 1.09-12.10). Our results add longitudinal data on blood Alu and LINE-1 methylation and cancer, and potentially contribute to their use as early-detection biomarkers. Future larger studies are needed and should account for the interval between blood sample collection and cancer diagnosis.


Signal processing for metagenomics: extracting information from the soup.
Traditionally, studies in microbial genomics have focused on single-genomes from cultured species, thereby limiting their focus to the small percentage of species that can be cultured outside their natural environment. Fortunately, recent advances in high-throughput sequencing and computational analyses have ushered in the new field of metagenomics, which aims to decode the genomes of microbes from natural communities without the need for cultivation. Although metagenomic studies have shed a great deal of insight into bacterial diversity and coding capacity, several computational challenges remain due to the massive size and complexity of metagenomic sequence data. Current tools and techniques are reviewed in this paper which address challenges in 1) genomic fragment annotation, 2) phylogenetic reconstruction, 3) functional classification of samples, and 4) interpreting complementary metaproteomics and metametabolomics data. Also surveyed are important applications of metagenomic studies, including microbial forensics and the roles of microbial communities in shaping human health and soil ecology.


Maternal smoking and the risk of orofacial clefts: Susceptibility with NAT1 and NAT2 polymorphisms.
Orofacial clefts are etiologically heterogeneous malformations. One probable cause is maternal smoking during pregnancy. The effect of maternal smoking may be modified by genes involved in biotransformation of toxic compounds derived from tobacco. We investigated whether polymorphic variants of fetal acetyl-N-transferases 1 (NAT1) and 2 (NAT2) interact with maternal cigarette smoking during early pregnancy to increase the risk of delivering an infant with an orofacial cleft. In a California population-based case-control study, we genotyped 421 infants born with an isolated cleft and 299 nonmal-formed controls for 2 NAT1 and 3 NAT2 single nucleotide polymorphisms Although smoking was independently associated with increased risks for both isolated cleft lip +/- cleft palate and isolated cleft palate, no independent associations were found for NAT1 1088 or 1095 genotypes or for NAT2 acetylator status. However, the infant NAT1 1088 and 1095 polymorphisms were strongly associated with the risk of clefts among smoking mothers; infants with NAT1 1088 genotype AA versus TT (odds ratio [OR] = 3.9; 95% confidence interval = 1.1-17.2) and with NAT1 1095 genotype AA versus CC (OR = 4.2; 1.2-18.0). Infant NAT2 acetylator status did not appreciably affect susceptibility of the fetus to the teratogenic effects of maternal smoking. Our results suggest that maternal smoking during pregnancy may increase risk for orofacial clefts particularly among smokers whose fetuses have polymorphic variants of NAT1, an enzyme involved in phase II detoxification of tobacco smoke constituents.


Plasma testosterone correlates with morph type across breeding substages in male white-throated Sparrows.
White-throated sparrows (Zonotrichia albicollis) exhibit a genetic polymorphism that affects plumage and behavior in both sexes. White-striped morphs are more territorially aggressive, whereas tan-striped morphs provision nestlings at a higher rate. We investigated testosterone physiology in this species in an effort to understand hormonal mechanisms for the observed differences in aggression and parental care between the morphs. We found a small but significant difference in plasma testosterone between free-living white-striped and tan-striped males over the course of the breeding season. This difference correlates with previously observed differences in aggressive behavior and suggests that testosterone may mediate these differences. Testosterone remained higher in white-striped males relative to tan-striped males when males were provisioning nestlings and fledglings. Thus, testosterone may also contribute to the relatively reduced levels of parental care exhibited by white-striped males. In contrast to males, plasma testosterone did not differ between free-living white-striped and tan-striped females, which suggests that testosterone does not mediate differences in aggression between female morphs. Injection with gonadotropin-releasing hormone led to greater testosterone secretion in both captive and free-living males and captive females but did not differ by morph. Therefore, we conclude that differences in plasma testosterone between the morphs are due to differences in testosterone regulation upstream of the pituitary.


Thermoregulation, Fluid Balance, and Sweat Losses in American Football Players.
Numerous studies have reported on the thermoregulation and hydration challenges athletes face in team and individual sports during exercise in the heat. Comparatively less research, however, has been conducted on the American Football player. Therefore, the purpose of this article is to review data collected in laboratory and field studies and discuss the thermoregulation, fluid balance, and sweat losses of American Football players. American Football presents a unique challenge to thermoregulation compared with other sports because of the encapsulating nature of the required protective equipment, large body size of players, and preseason practice occurring during the hottest time of year. Epidemiological studies report disproportionately higher rates of exertional heat illness and heat stroke in American Football compared with other sports. Specifically, larger players (e.g., linemen) are at increased risk for heat ailments compared with smaller players (e.g., backs) because of greater body mass index, increased body fat, lower surface area to body mass ratio, lower aerobic capacity, and the stationary nature of the position, which can reduce heat dissipation. A consistent finding across studies is that larger players exhibit higher sweating rates than smaller players. Mean sweating rates from 1.0 to 2.9 L/h have been reported for college and professional American Football players, with several studies reporting 3.0 L/h or more in some larger players. Sweat sodium concentration of American Football players does not seem to differ from that of athletes in other sports; however, given the high volume of sweat loss, the potential for sodium loss is higher in American Football than in other sports. Despite high sweating rates with American Football players, the observed disturbances in fluid balance have generally been mild (mean body mass loss ≤2 %). The majority of field-based studies have been conducted in the northeastern part of the United States, with limited studies in different geographical regions (i.e., southeast) of the United States. Further, there have been a limited number of studies examining body core temperature of American Football players during preseason practice, especially at the high school level. Future field-based research in American Football with various levels of competition in hotter geographical regions of the United States is warranted.


Predicting the outcome of surgery for the proximal interphalangeal joint in Dupuytren's disease.
We prospectively studied the outcome of limited Dupuytren's fasciectomy, in combination with joint release if necessary, for disease involving 49 proximal interphalangeal joints (PIPJs) to identify factors that predispose to recurrent PIPJ contracture. Thirty-seven patients were treated over a 4-year period. The flexion contracture of the PIPJ was measured before surgery, immediately after surgery, and at more than 1 year after surgery. A mean preoperative flexion contracture of 67 degrees +/- 22 degrees was corrected to 6 degrees +/- 10 degrees at the time of surgery and 25 degrees +/- 25 degrees at the follow-up evaluation. There was a positive correlation between the severity of the preoperative flexion contracture and recurrent deformity, with a preoperative contracture greater than 60 degrees leading to significantly worse outcome. Incomplete correction of PIPJ flexion contracture during surgery and poor postoperative compliance with therapy were also associated with worse recurrent joint contractures. The digit involved and the necessity for joint release did not significantly affect outcome. In the absence of recurrent Dupuytren's disease, severe preoperative deformity, incomplete correction at surgery, and noncompliance with therapy predispose patients to worse PIPJ contracture. Prognostic II.


Vitamin C protects carboplatin-exposed oocytes from meiotic failure.
CBP (carboplatin) is a second-generation chemotherapeutic drug of platinum compound commonly applied in the treatment of sarcomas and germ cell tumours. Although it is developed to replace cisplatin, which has been proven to have a variety of side effects during cancer treatment, CBP still exhibits a certain degree of toxicity including neurotoxicity, nephrotoxicity, hematotoxicity and myelosuppression. However, the underlying mechanisms regarding how CBP influences the female reproductive system especially oocyte quality have not yet been fully determined. Here, we report that CBP exposure led to the oocyte meiotic defects by impairing the dynamics of the meiotic apparatus, leading to a remarkably aberrant spindle organisation, actin polymerisation and mitochondrial integrity. Additionally, CBP exposure caused compromised sperm binding and fertilisation potential of oocytes by due to an abnormal distribution of cortical granules and its component ovastacin. More importantly, we demonstrated that vitamin C supplementation prevented meiotic failure induced by CBP exposure and inhibited the increase in ROS levels, DNA damage accumulation and apoptotic incidence. Taken together, our findings demonstrate the toxic effects of CBP exposure on oocyte development and provide a potential effective way to improve the quality of CBP-exposed oocytes in vitro.


Injectable cartilage using polyethylene oxide polymer substrates.
This study demonstrates that polyethylene oxide gels, which are biocompatible and biodegradable synthetic polymers, can be utilized for the encapsulation of isolated chondrocytes and maintenance of three-dimensional spatial support for new tissue development. Chondrocytes isolated from the glenohumeral and humeroradioulnar joints of a calf were added to a 20% polyethylene oxide solution in Ham's F-12 medium to generate a final cellular density of 10 x 10(6)/mL. The polymer-chondrocyte constructs were injected through a 22-gauge needle in 500-microliters aliquots subcutaneously in 12 nude mice and incubated for 6 and 12 weeks in vivo. Histologic and biochemical analyses including deoxyribonucleic acid and glycosaminoglycan quantitative analyses confirmed the presence of actively proliferating chondrocytes with production of a well-formed cartilaginous matrix in the transplanted samples. Control specimens from eight implantation sites consisting of chondrocytes alone or polyethylene oxide substrates did not demonstrate any gross or histologic evidence of neocartilage formation. These findings demonstrate the potential use of an injectable and moldable polymer substrate that can support cell proliferation and matrix synthesis after subcutaneous transplantation for neocartilage generation. The use of functional biologic tissue substitutes may serve as an alternative solution to current methods of augmentation or reconstruction of structural craniofacial contour deformities.


Macroevolution of complex retroviruses.
Retroviruses can leave a "fossil record" in their hosts' genomes in the form of endogenous retroviruses. Foamy viruses, complex retroviruses that infect mammals, have been notably absent from this record. We have found an endogenous foamy virus within the genomes of sloths and show that foamy viruses were infecting mammals more than 100 million years ago and codiverged with their hosts across an entire geological era. Our analysis highlights the role of evolutionary constraint in maintaining viral genome structure and indicates that accessory genes and mammalian mechanisms of innate immunity are the products of macroevolutionary conflict played out over a geological time scale.


Molecular insight into how HIV-1 Vpr protein impairs cell growth through two genetically distinct pathways.
Vpr, a small HIV auxiliary protein, hijacks the CUL4 ubiquitin ligase through DCAF1 to inactivate an unknown cellular target, leading to cell cycle arrest at the G(2) phase and cell death. Here we first sought to delineate the Vpr determinants involved in the binding to DCAF1 and to the target. On the one hand, the three α-helices of Vpr are necessary and sufficient for binding to DCAF1; on the other hand, nonlinear determinants in Vpr are required for binding to the target, as shown by using protein chimeras. We also underscore that a SRIG motif conserved in the C-terminal tail of Vpr proteins from HIV-1/SIVcpz and HIV-2/SIVsmm lineages is critical for G(2) arrest. Our results suggest that this motif may be predictive of the ability of Vpr proteins from other SIV lineages to mediate G(2) arrest. We took advantage of the characterization of a subset of G(2) arrest-defective, but DCAF1 binding-proficient mutants, to investigate whether Vpr interferes with cell viability independently of its ability to induce G(2) arrest. These mutants inhibited cell colony formation in HeLa cells and are cytotoxic in lymphocytes, unmasking a G(2) arrest-independent cytopathic effect of Vpr. Furthermore these mutants do not block cell cycle progression at the G(1) or S phases but trigger apoptosis through caspase 3. Disruption of DCAF1 binding restored efficiency of colony formation. However, DCAF1 binding per se is not sufficient to confer cytopathicity. These data support a model in which Vpr recruits DCAF1 to induce the degradation of two host proteins independently required for proper cell growth.


Correlation of Nitrogen Sorption and Confocal Laser Scanning Microscopy for the Analysis of Amino Group Distributions on Mesoporous Silica.
Aminopropylalkoxysilanes are frequently used for the functionalization of mesoporous silica. The analysis of amino group distributions on arrays of silica nanochannels by a combination of nitrogen sorption and confocal laser scanning microscopy provides valuable insight into the mechanisms underlying the interaction of these silanes with mesoporous silica surfaces. Tendencies towards external surface functionalization, non-uniform distribution in the pores, and hydrolysis of the silica framework are shown to depend to a large extent on the mobility of the aminopropylalkoxysilane molecules, which can be adjusted by the number and type of alkoxy groups.


Alcohol potently modulates climbing fiber-->Purkinje neuron synapses: role of metabotropic glutamate receptors.
Consumption of alcoholic beverages produces alterations in motor coordination and equilibrium that are responsible for millions of accidental deaths. Studies indicate that ethanol produces these alterations by affecting the cerebellum, a brain region involved in the control of motor systems. Purkinje neurons of the cerebellar cortex have been shown to be particularly important targets of ethanol. However, its mechanism of action at these neurons is poorly understood. We hypothesized that ethanol could modulate Purkinje neuron function by altering the excitatory input provided by the climbing fiber from the inferior olive, which evokes a powerful all-or-none response denoted as the complex spike. To test this hypothesis, we performed whole-cell patch-clamp electrophysiological and Ca2+ imaging experiments in acute slices from rat cerebella. We found that ethanol potently inhibits the late phase of the complex spike and that this effect is the result of inhibition of type-1 metabotropic glutamate receptor-dependent responses at the postsynaptic level. Moreover, ethanol inhibited climbing fiber long-term depression, a form of synaptic plasticity that also depends on activation of these metabotropic receptors. Our findings identify the climbing fiber-->Purkinje neuron synapse as an important target of ethanol in the cerebellar cortex and indicate that ethanol significantly affects cerebellar circuits even at concentrations as low as 10 mm (legal blood alcohol level in the United States is below 0.08 g/dl = 17 mm).