HIV drug resistance threshold survey using specimens from voluntary counselling and testing sites in Hanoi, Vietnam.
In countries where antiretroviral therapy has been available or is being rapidly expanded, the World Health Organization (WHO) recommends surveillance for transmitted HIV drug resistance (HIVDR) by threshold surveillance methods using specimens from antenatal clinics or voluntary counselling and testing (VCT) sites. The aim of this study was to implement the HIVDR threshold survey in VCT sites in Vietnam, where HIV prevalence is high. Estimating transmitted resistance in the infected population will enable the appropriateness of current antiretroviral drug regimens to be assessed and will inform plans for future HIVDR surveillance. Consecutive blood specimens were collected from 70 newly diagnosed HIV-positive clients 18-24 years of age at two sites in Hanoi, Vietnam. Informed consent and serum specimens were obtained from each eligible client, with serum frozen at -70 degrees C until shipping to Thailand for resistance testing using the TruGene system. From February until August 2006, 559 clients were eligible to participate in this survey. Of the 535 clients (95.7%) who agreed to participate, 70 (13%) were HIV-positive and were included in the survey. Of the 70 specimens sent for genotyping, 52 consecutive samples were amplified, 49 of which could be genotyped. Only 1 of 49 genotyped specimens had mutations associated with drug resistance (L74V and Y181C) in the reverse transcriptase gene, indicating that the prevalence of transmitted HIVDR to all drugs and drug classes evaluated was <5%. The prevalence of transmitted HIVDR was low in Hanoi as determined using threshold surveillance methods. The Ministry of Health plans to repeat this survey methodology in one more province and to confirm these findings by expanded HIVDR surveillance.


Internet Gaming Disorder Among Slovenian Primary Schoolchildren: Findings From a Nationally Representative Sample of Adolescents.
Background and aims Since the inclusion of Internet Gaming Disorder (IGD) in the latest (fifth) edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) as a tentative disorder, a few psychometric screening instruments have been developed to assess IGD, including the 9-item Internet Gaming Disorder Scale - Short-Form (IGDS9-SF) - a short, valid, and reliable instrument. Methods Due to the lack of research on IGD in Slovenia, this study aimed to examine the psychometric properties of the IGDS9-SF in addition to investigating the prevalence rates of IGD in a nationally representative sample of eighth graders from Slovenia (N = 1,071). Results The IGDS9-SF underwent rigorous psychometric scrutiny in terms of validity and reliability. Construct validation was investigated with confirmatory factor analysis to examine the factorial structure of the IGDS9-SF and a unidimensional structure appeared to fit the data well. Concurrent and criterion validation were also investigated by examining the association between IGD and relevant psychosocial and game-related measures, which warranted these forms of validity. In terms of reliability, the Slovenian version IGDS9-SF obtained excellent results regarding its internal consistency at different levels, and the test appears to be a valid and reliable instrument to assess IGD among Slovenian youth. Finally, the prevalence rates of IGD were found to be around 2.5% in the whole sample and 3.1% among gamers. Discussion and conclusion Taken together, these results illustrate the suitability of the IGDS9-SF and warrants further research on IGD in Slovenia.


Endometrial side population cells: potential adult stem/progenitor cells in endometrium.
Uterine endometrium is one of the most important organs for species preservation. However, the physiology of human endometrium remains poorly understood, because the human endometrium undergoes rapid and large changes during each menstrual cycle and it is very difficult to investigate human endometrium as one organ. This remarkable regenerative capacity of human endometrium strongly suggests the existence of adult stem cells, and physiology of endometrium cannot be explained without adult stem cells. Therefore, investigating endometrial stem/progenitor cells should lead to a breakthrough in understanding the normal endometrial physiology and the pathophysiology of endometrial neoplastic disorders, such as endometriosis and endometrial cancer. Several cell populations have been discovered as putative endometrial stem/progenitor cells. Emerging evidence reveals that the endometrial side population (SP) is one of the potential endometrial stem/progenitor populations. Of all the endometrial stem/progenitor cell candidates, the endometrial SP (ESP) is best investigated in vitro and in vivo, and has the largest number of references. In this review, we provide an overview of the accumulating evidence for the ESP cells, both directly from human endometria and from cultured endometrial cells. Furthermore, SP cells are compared to other potential stem/progenitor cells, and we discuss their stem cell properties. We also discuss the difficulties and unsolved issues in endometrial stem cell biology.


Innovative surgical endoscopes in video-assisted thoracic surgery.
In the past three decades, rod lens endoscopes had facilitated the development and wide spread applications of video-assisted thoracic surgery (VATS). With the rise of uniportal VATS in recent years, innovations in surgical instruments should once again complement the advancement in surgical technique. While articulated flexible endoscopes have expand the field of view, and can alter viewing direction with minimal maneuvers, they still suffer from problems like trocar crowding and interference with other instruments. Magnetic anchored endoscopes, on the other hand, may provide unique benefits to VATS by replacing the endoscope rigid rod body with magnetic linkage, thus overcoming the challenge of port crowding in single incision surgery. Most magnetic anchored endoscopes reported in literature are not designed for thoracic surgeries. Many of these designs do not allow tilting of endoscopic view, rely on micromotors for actuation, or are ergonomically unfit to be operated within the spatial constraints seen in VATS application. Considering these limitations, we have designed two novel magnetic anchored and steered endoscopes targeted for uniportal VATS. Both designs could be wirelessly actuated by magnetic interaction. One has a silicone rubber formed soft body for compactness, lightweight and safety, while another is a 40 mm long capsule optimized for VATS spatial constraints.


Effect of inhaled essential oils on mental exhaustion and moderate burnout: a small pilot study.
The objective of this pilot study was to determine the effectiveness of a mixture of essential oils (peppermint, basil, and helichrysum) on mental exhaustion, or moderate burnout (ME/MB) using a personal inhaler. This was a randomized, controlled, double-blind pilot study. Data were collected 3 times a day for 3 weeks (Monday-Friday). The first week was baseline for both groups, the second week was intervention (aromatherapy or placebo), and the third week was washout. Participants used a personal inhaler at home or at work. The subjects comprised a convenience sample of 13 women and 1 man who each had self-assessed ME/MB. Participants were randomized to receive a personal inhaler containing either a mixture of essential oils or rose water (as used in Indian cooking). The outcome measures were a 0-10 scale with 10=worst feeling of burnout, 0=no feeling of burnout. There was a qualitative questionnaire rating aroma and a questionnaire listing perceived stressors. While both groups had a reduction in perception of ME/MB, the aromatherapy group had a much greater reduction. The results suggest that inhaling essential oils may reduce the perceived level of mental fatigue/burnout. Further research is warranted.


Risk factors for repetitive ileocolic resection in patients with Crohn's disease: results of an observational cohort study.
Surgical recurrence rates among patients with Crohn's disease with ileocolic resection (ICR) remain high, and factors predicting surgical recurrence remain controversial. We aimed to identify risk and protective factors for repetitive ICRs among patients with Crohn's disease in a large cohort of patients. Data on 305 patients after first ICR were retrieved from our cross-sectional and prospective database (median follow-up: 15 yr [0-52 yr]). Data were compared between patients with 1 (ICR = 1, n = 225) or more than 1 (ICR >1, n = 80) resection. Clinical phenotypes were classified according to the Montreal Classification. Gender, family history of inflammatory bowel disease, smoking status, type of surgery, immunomodulator, and biological therapy before, parallel to and after first ICR were analyzed. The mean duration from diagnosis until first ICR did not differ significantly between the groups, being 5.93 ± 7.65 years in the ICR = 1 group and 5.36 ± 6.35 years in the ICR >1 group (P = 0.05). Mean time to second ICR was 6.7 ± 5.74 years. In the multivariate logistic regression analysis, ileal disease location (odds ratio [OR], 2.42; 95% confidence interval [CI], 1.02-5.78; P = 0.05) was a significant risk factor. A therapy with immunomodulators at time of or within 1 year after first ICR (OR, 0.23; 95% CI, 0.09-0.63; P < 0.01) was a protective factor. Neither smoking (OR, 1.16; 95% CI, 0.66-2.06) nor gender (male OR, 0.85; 95% CI, 0.51-1.42) or family history (OR, 1.68; 95% CI, 0.84-3.36) had a significant impact on surgical recurrence. Immunomodulators have a protective impact regarding surgical recurrence after ICR. In contrast, ileal disease location constitutes a significant risk factor for a second ICR.


Multiple differentiation capacity of STRO-1+/CD146+ PDL mesenchymal progenitor cells.
Although mesenchymal progenitor cells can be isolated from periodontal ligament (PDL) tissues using stem cell markers STRO-1 and CD146, the proportion of these cells that have the capacity to differentiate into multiple cell lineages remains to be determined. This study was designed to quantify the proportions of primary human PDL cells that can undergo multilineage differentiation and to compare the magnitude of these capabilities relative to bone marrow-derived mesenchymal stem cells (MSCs) and parental PDL (PPDL) cells. PDL mesenchymal progenitor (PMP) cells were isolated from PPDL cells using the markers STRO-1 and CD146. The colony-forming efficiency and multilineage differentiation potential of PMP, PPDL, and MSCs under chondrogenic, osteogenic, and adipogenic conditions were determined. Flow cytometry revealed that on average 2.6% of PPDL cells were STRO-1(+)/CD146(+), whereas more than 63% were STRO-1(-)/CD146(-). Colony-forming efficiency of STRO-1(+)/CD146(+) PMP cells (19.3%) and MSCs (16.7%) was significantly higher than that of PPDL cells (6.8%). Cartilage-specific genes, early markers of osteoblastic differentiation, and adipogenic markers were significantly upregulated under appropriate conditions in PMP cells and MSCs compared to either their noninduced counterparts or induced PPDL cells. Consistent with these findings, immunohistochemistry revealed substantial accumulation of cartilaginous macromolecules, mineralized calcium nodules, and lipid vacuoles under chondrogenic, osteogenic, or adipogenic conditions in PMP and MSC cultures, respectively, compared to noninduced controls or induced PPDL cells. Thus STRO-1(+)/CD146(+) PMP cells demonstrate multilineage differentiation capacity comparable in magnitude to MSCs and could potentially be utilized for regeneration of the periodontium and other tissues.


Lycium barbarum polysaccharides inhibit proliferation and migration of bladder cancer cell lines BIU87 by suppressing Pi3K/AKT pathway.
The aim of this study was to verify whether Lycium barbarum polysaccharides inhibits proliferation and migration of BIU87 cells through Pi3K/AKT pathway. Different concentrations of Lycium barbarum polysaccharides were used to incubate with BIU87cells. LY-294002 and IGF-1 were used to inhibit and activate Pi3K/AKT pathway respectively. MTT were used to investigate the proliferation of BIU87cells. Transwell chambers and wound healing were used to test the migratory ability of BIU87cells. Western blotting were used to investigate the expressions of P21,P27,MMP-2, MMP-9, AKT and p-AKT in BIU87cells. Compared with the control group, the proliferation and migration of BIU87cells and the expression of p-AKT were significantly decreased in the study group; the inhibitory effect of the downregulation of p-AKT by LY-294002on the induction of BIU87cells proliferation and migration was identical to that of Lycium barbarum polysaccharides; upregulation of p-AKT by IGF-1 reversed the Lycium barbarum polysaccharides-induced inhibition of BIU87cells dedifferentiation. In conclusion, LBP inhibits the proliferation and migration of BIU87 cells by suppressing Pi3K/AKT signaling pathway.


Interfacing enzymes with silicon nanocrystals through the thiol-ene reaction.
This study reports the preparation of functional bioinorganic hybrids, through application of the thiol-ene reaction, that exhibit catalytic activity and photoluminescent properties from enzymes and freestanding silicon nanocrystals. Thermal hydrosilylation of 1,7-octadiene and alkene-terminated poly(ethylene oxide)methyl ether with hydride-terminated silicon nanocrystals afforded nanocrystals functionalized with alkene residues and poly(ethylene oxide) moieties. These silicon nanocrystals were conjugated with representative enzymes through the photochemical thiol-ene reaction to afford bioinorganic hybrids that are dispersible and photostable in buffer, and that exhibit photoluminescence (λmax = 630 nm) and catalytic activity. They were characterized using Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), dynamic light scattering analysis (DLS), absorption spectroscopy, steady-state and time-resolved photoluminescence spectroscopy, and pertinent enzyme activity assays. The general derivatization approach presented for interfacing enzymes with biocompatible silicon nanocrystals has far reaching implications for many applications ranging from sensors to therapeutic agents. The bioinorganic hybrids presented herein have potential applications in the chemical detection of nitrophenyl esters and urea. They can also be employed in enzyme-based theranostics as they combine long-lived silicon nanocrystal photoluminescence with substrate-specific enzymatic activity.


Reward processing by the lateral habenula in normal and depressive behaviors.
The brain reward circuit has a central role in reinforcing behaviors that are rewarding and preventing behaviors that lead to punishment. Recent work has shown that the lateral habenula is an important part of the reward circuit by providing 'negative value' signals to the dopaminergic and serotonergic systems. Studies have also suggested that dysfunction of the lateral habenula is associated with psychiatric disorders, including major depression. Here, we discuss insights gained from neuronal recordings in monkeys regarding how the lateral habenula processes reward-related information. We then highlight recent optogenetic experiments in rodents addressing normal and abnormal functions of the habenula. Finally, we discuss how deregulation of the lateral habenula may be involved in depressive behaviors.


Structural basis for androgen specificity and oestrogen synthesis in human aromatase.
Aromatase cytochrome P450 is the only enzyme in vertebrates known to catalyse the biosynthesis of all oestrogens from androgens. Aromatase inhibitors therefore constitute a frontline therapy for oestrogen-dependent breast cancer. In a three-step process, each step requiring 1 mol of O(2), 1 mol of NADPH, and coupling with its redox partner cytochrome P450 reductase, aromatase converts androstenedione, testosterone and 16alpha-hydroxytestosterone to oestrone, 17beta-oestradiol and 17beta,16alpha-oestriol, respectively. The first two steps are C19-methyl hydroxylation steps, and the third involves the aromatization of the steroid A-ring, unique to aromatase. Whereas most P450s are not highly substrate selective, it is the hallmark androgenic specificity that sets aromatase apart. The structure of this enzyme of the endoplasmic reticulum membrane has remained unknown for decades, hindering elucidation of the biochemical mechanism. Here we present the crystal structure of human placental aromatase, the only natural mammalian, full-length P450 and P450 in hormone biosynthetic pathways to be crystallized so far. Unlike the active sites of many microsomal P450s that metabolize drugs and xenobiotics, aromatase has an androgen-specific cleft that binds the androstenedione molecule snugly. Hydrophobic and polar residues exquisitely complement the steroid backbone. The locations of catalytically important residues shed light on the reaction mechanism. The relative juxtaposition of the hydrophobic amino-terminal region and the opening to the catalytic cleft shows why membrane anchoring is necessary for the lipophilic substrates to gain access to the active site. The molecular basis for the enzyme's androgenic specificity and unique catalytic mechanism can be used for developing next-generation aromatase inhibitors.


Weekly variations of viruses and heterotrophic nanoflagellates and their potential impact on bacterioplankton in shallow waters of the central Red Sea.
Bacterioplankton play a pivotal role in marine ecosystems. However, their temporal dynamics and underlying control mechanisms are poorly understood in tropical regions such as the Red Sea. Here, we assessed the impact of bottom-up (resource availability) and top-down (viruses and heterotrophic nanoflagellates) controls on bacterioplankton abundances by weekly sampling a coastal central Red Sea site in 2017. We monitored microbial abundances by flow cytometry together with a set of environmental variables including temperature, salinity, dissolved organic and inorganic nutrients and chlorophyll a. We distinguished five groups of heterotrophic bacteria depending on their physiological properties relative nucleic acid content, membrane integrity and cell-specific respiratory activity, two groups of Synechococcus cyanobacteria and three groups of viruses. Viruses controlled heterotrophic bacteria for most of the year, as supported by a negative correlation between their respective abundances and a positive one between bacterial mortality rates and mean viral abundances. On the contrary, heterotrophic nanoflagellates abundance covaried with that of heterotrophic bacteria. Heterotrophic nanoflagellates showed preference for larger bacteria from both the high and low nucleic acid content groups. Our results demonstrate that top-down control is fundamental in keeping heterotrophic bacterioplankton abundances low (< 5 × 10 5 cells mL-1) in Red Sea coastal waters.


Colorimetric ultrasensitive detection of DNA based on the intensity of gold nanoparticles with dark-field microscopy.
We present an ultrasensitive colorimetric nucleic acid assay based on the intensity of gold nanoparticles (Au NPs) using dark field microscopy. In the absence of target DNA, two hairpin-like DNA strands with protruding single-stranded DNA (ssDNA) can be absorbed onto the Au NP surface via non-covalent interactions between the exposed nitrogen bases of ssDNA and Au NPs, which inhibits Au NPs from aggregating in a high concentration of salt media, while in the presence of target DNA, two hairpin DNA strands hybridize with target DNA to form double-stranded DNA (dsDNA). After hybridization chain reaction (HCR) amplification, rigid dsDNA polymers are formed, which results in serious Au NP aggregation in the salt environment. By measuring the intensity change of yellow and red dots on a dark-field image, the concentration of target DNA can be accurately quantified with a limit of detection (LOD) of 66 fM.


Maintenance therapy with second generation antipsychotics for bipolar disorder - A systematic review and meta-analysis.
Second generations antipsychotics (SGA) are frequently used for maintenance treatment in bipolar disorder. We systematically reviewed the efficacy and long-term effects of treatment with SGA, regardless of treatment strategy (SGA administered either as monotherapy or as adjunctive therapy), in comparison to placebo, lithium or valproate. Primary outcomes were relapses (mood episode recurrence) and discontinuation. Clinical studies were identified through database searching in PubMed, Embase, PsychInfo and Cochrane Library and critically appraised based on the Cochrane Handbook. Full data extraction of raw data was performed and analyzed with meta-analyses, and level of evidence graded using GRADE. Only randomized controlled studies (RCT) and observational studies were included, with a minimum follow-up of 6 months. Comparators used were restricted to placebo, lithium, valproate or other anti-epileptic drugs. We identified 15 RCTs on SGA in bipolar disorder with follow-up-time of 6 months up to 2 years, and one observational study reporting long-term effects of up to 4 years. A total of 6142 patients were included in the randomized trials. No long-term RCTs beyond 2 years follow-up was identified. All RCTs except for one included patients with bipolar disorder type I only. All RCTs except for two included patients pre-stabilized on the drug under investigation prior to randomization (enrichment design). For SGA as adjunctive therapy to lithium or valproate, meta-analyses showed that treatment with either aripiprazole (RR: 0.65, 95% CI 0.50-0.85), quetiapine (RR: 0.38, 95% CI 0.32-0.46) or ziprasidone (RR: 0.62, 95% CI 0.40-0.96) reduced the overall risk of relapses in patients that had responded during the stabilization phase. Adjunctive therapy with quetiapine was the only drug that reduced both manic and depressive episodes. For SGA as monotherapy, only quetiapine was shown to be better than lithium/ valproate for both manic and depressive relapses, but only for patients stabilized on quetiapine during the acute phase. As monotherapy, olanzapine, quetiapine and risperidone were shown to be superior to placebo in reducing the overall risk of relapses. There were considerable limitations to the evidence base of maintenance treatment with SGA in bipolar disorder. Most studies used stabilized patients, i.e. enrichment design (selection bias), had considerable dropout levels (attrition bias), and variable degree of reporting bias. No long-term RCT data on efficacy is available beyond 2 years, and almost all studies are on bipolar disorder type I patients only. Despite these limitations, we elucidate quantitative findings from meta-analyses conducted on the randomized trials published on the topic.


Neurological deficits and glycosphingolipid accumulation in saposin B deficient mice.
Saposin B derives from the multi-functional precursor, prosaposin, and functions as an activity enhancer for several glycosphingolipid (GSL) hydrolases. Mutations in saposin B present in humans with phenotypes resembling metachromatic leukodystrophy. To gain insight into saposin B's physiological functions, a specific deficiency was created in mice by a knock-in mutation of an essential cysteine in exon 7 of the prosaposin locus. No saposin B protein was detected in the homozygotes (B-/-) mice, whereas prosaposin, and saposins A, C and D were at normal levels. B-/- mice exhibited slowly progressive neuromotor deterioration and minor head tremor by 15 months. Excess hydroxy and non-hydroxy fatty acid sulfatide levels were present in brain and kidney. Alcian blue positive (sulfatide) storage cells were found in the brain, spinal cord and kidney. Ultrastructural analyses showed lamellar inclusion material in the kidney, sciatic nerve, brain and spinal cord tissues. Lactosylceramide (LacCer) and globotriaosylceramide (TriCer) were increased in various tissues of B-/- mice supporting the in vivo role of saposin B in the degradation of these lipids. CD68 positive microglial cells and activated GFAP positive astrocytes showed a proinflammatory response in the brains of B-/- mice. These findings delineate the roles of saposin B for the in vivo degradation of several GSLs and its primary function in maintenance of CNS function. B-/- provide a useful model for understanding the contributions of this saposin to GSL metabolism and homeostasis.


Nutritional problems in urban slum children.
Nutritional problems like protein energy malnutrition (PEM), anemia and vitamin A deficiency continue to plague a large proportion of Indian children. The diets and nutritional status of urban slum children in India is far away from being satisfactory. The nutritional status of slum children is worst amongst all urban groups and is even poorer than the rural average. Urban migration has not provided them salvation from poverty and undernutrition. Another distressing feature is the lack of any significant improvement over the years in this population. Most common causes of malnutrition include faulty infant feeding practices, impaired utilization of nutrients due to infections and parasites, inadequate food and health security, poor environmental conditions and lack of proper child care practices. High prevalence of malnutrition among young children is also due to lack of awareness and knowledge regarding their food requirements and absence of a responsible adult care giver. With increasing urban migration in the years ahead, the problem of malnutrition in urban slums will also acquire increasing dimension unless special efforts are initiated to mitigate the health and nutrition problems of the urban poor. Improving nutritional status of urban poor requires a more direct, more focused, and more integrated strategy.


Indigenous healing practice: ayahuasca. Opening a discussion.
This essay frames an invitation to pastoral counselors and pastoral theologians to examine connections and perhaps interactions between themselves and traditional shamanic healers who use ayahuasca in their healing ceremonies. Indigenous people in South America have used ayahuasca for centuries, and the ritual has become common among the mestizo populations in urban areas of the Amazon, particularly as a curing ritual for drug addiction (Dobkin de Rios, 1970; Moir, 1998). Like peyote in the United States (Calabrese, 1997) ayahuasca use amongst the indigenous people of the Amazon is a form of cultural psychiatry. A review of the literature reveals very little commentary or discussion of shamanic practice in Pastoral Counseling (Pastoral Theology). The scant literature identifies an antithetical relationship at best. The current authors wonder about the possibility of to including shamanic practices in the context of pastoral counseling? This essay seeks to provide some basic information about the ritual use of ayahuasca and to offer a rationale for pastoral counselors to engage in a dialogue about its utility.


Developing meaningful client-dietitian relationships in the chronic disease context: An exploration of dietitians' perspectives.
Meaningful client-dietitian relationships are central to effective dietetic practice. The chronic disease management setting provides an opportunity to examine what is meaningful and how these relationships are constructed, because the dietitian and client generally have multiple interactions over an extended period of time. This study aimed to explore dietitians' perspectives of how they develop meaningful relationships with clients managing lifestyle-related chronic diseases. Study design and analysis were guided by Charmaz's constructivist grounded theory. Dietitians working in Australia with clients managing chronic diseases were recruited through initial, snowball and theoretical sampling. Online videoconference and telephone semi-structured interviews were conducted. Recorded interview transcripts were analysed using repeated reviews comprising initial, focused and theoretical coding and memoing. Twenty-two dietitians were recruited. A conceptual model developed from the data showed the dietitian's role in developing the client-dietitian relationship is complex. Key elements were identified and described as 'Sensing a Professional Chemistry', and the dietitian's skills in 'Balancing Professional and Social Relationships' and 'Managing Tension with Competing Influences'. Influences were categorised as relating to the client and dietitian as individuals (eg, their values), their support network and external contextual factors (eg, working with interpreters). Developing relationships with clients in the chronic disease context appears complex due to the dietitian's role of managing multiple interrelated elements and influential factors simultaneously. To deepen understanding, research should explore clients' perspectives of relationship development and how knowledge of practitioner-client relationships in other disciplines may be utilised to enhance dietetic service delivery.


Integration of hybrid silicon lasers and electroabsorption modulators.
We present an integration platform based on quantum well intermixing for multi-section hybrid silicon lasers and electroabsorption modulators. As a demonstration of the technology, we have fabricated discrete sampled grating DBR lasers and sampled grating DBR lasers integrated with InGaAsP/InP electroabsorption modulators. The integrated sampled grating DBR laser-modulators use the as-grown III-V bandgap for optical gain, a 50 nm blue shifted bandgap for the electrabosprtion modulators, and an 80 nm blue shifted bandgap for low loss mirrors. Laser continuous wave operation up to 45 ?C is achieved with output power >1.0 mW and threshold current of <50 mA. The modulator bandwidth is >2GHz with 5 dB DC extinction.


Regulation of Human UGT2B15 and UGT2B17 by miR-376c in Prostate Cancer Cell Lines.
Given the prime importance of UDP-glucuronosyltransferase (UGT) 2B15 and UGT2B17 in inactivating testosterone and dihydrotestosterone, control of their expression and activity in the prostate is essential for androgen signaling homeostasis in this organ. Although several studies provide evidence of transcriptional control of UGT2B15 and UGT2B17 by various endogenous and exogenous compounds, potential post-transcriptional regulation of UGT2B15 and UGT2B17 by microRNAs (miRs) in prostate cancer cells has not been examined. The present study identified a putative miR-376c target site in the 3'-untranslated regions (UTRs) of both UGT2B15 and UGT2B17 mRNAs. In accordance with the possibility that this miRNA negatively regulates UGT2B15 and UGT2B17 expression, there is an inverse correlation in the levels of miR-376c and UGT2B15/UGT2B17 mRNAs in prostate cancer cell lines versus normal prostate tissue. In LNCaP cells, transfection of miR-376c mimics inhibited the glucuronidations of testosterone, 4-methylumbelliferone (a substrate of UGT2B15), and androsterone (a substrate of UGT2B17). miR-376c reduced both UGT2B15 and UGT2B17 mRNA and protein levels and the activity of luciferase reporters containing UGT2B15 or UGT2B17 3'-UTRs. This microRNA-mediated repression was significantly abrogated by mutating the miR-376c binding site in the 3'-UTRs of both UGTs. Collectively, these data indicate that the expression of UGT2B15 and UGT2B17 is negatively regulated by the binding of miR-376c to the 3'-UTRs of UGT2B15 and UGT2B17 in prostate cancer cells. This represents the first evidence for post-transcriptional regulation of UGT2B15 and UGT2B17 by miRNAs in prostate cancer cells and may have importance in regulating androgen receptor signaling.


Less Frequent and Less Severe Flu-Like Syndrome in Interferon Beta-1a Treated Multiple Sclerosis Patients with at Least One Allele Bearing the G>C Polymorphism at Position -174 of the IL-6 Promoter Gene.
One of the most common adverse event of interferon beta (IFNβ) therapy for multiple sclerosis is flu-like syndrome (FLS), which has been reportedly related to increased levels of cytokines such as interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Average cytokine levels can be affected by single nucleotide polymorphism in the gene promoter regions. To investigate whether IL-6 -174 G>C and TNF-α -376 G>A polymorphisms could be correlated to the incidence of FLS, and whether an anti-inflammatory/antipyretic therapy may influence FLS development, a prospective observational study was performed in 190 treatment naïve, multiple sclerosis patients who started IM IFNβ-1a 30mcg once weekly. The identification of IL-6 -174 G>C and TNF-α -376 G>A polymorphisms was achieved by performing an amplification-refractory mutation system. Serum IL-6 levels were measured using enzyme-linked immunosorbent assay in blood samples taken before therapy and then after the first and last IFNβ-1a injection of the follow-up. FLS-related symptoms were recorded by patients once per week during the first 12 weeks of therapy into a self-reported diary. We found that patients carrying at least one copy of the C allele at position -174 in the promoter of IL-6 gene produced lower levels of IL-6 and were less prone to develop FLS, which was also less severe. On the contrary, the polymorphism of TNF-α had no effect on FLS. Patients taking the first dose of anti-inflammatory/antipyretic therapy in the peri-injection period (within 1 hour) experienced a reduced FLS severity. In conclusion, the study of IL-6 -174 G>C polymorphism would allow the identification of patients lacking the C nucleotide on both alleles who are at risk of a more severe FLS, and may be addressed to a timely and stronger anti-inflammatory/antipyretic therapy for a more effective FLS prevention.


Lack of recognition of HLA class I mismatches outside α1/α2 domains by CD8+ alloreactive T lymphocytes: the HLA-B44 paradigm.
Transplantation with hematopoietic stem cells (HSC) from a donor with a single human leukocyte antigen (HLA) mismatch can be proposed to those patients lacking an HLA identical sibling donor or an unrelated donor matched for the HLA-A, -B, -C, DRB1, DQB1 loci. Incompatibilities at HLA classes I and II loci are associated with an increased risk of graft-versus-host disease (GVHD) and mortality, although no consensus exists yet on the relative importance of specific allele disparities on clinical outcome. Donor search algorithms are now complicated by the growing number of new HLA alleles, in particular those that differ outside the peptide-binding site of the HLA molecules. We report here an in vitro cellular assay to quantify CD8+CD137+ alloreactive cytotoxic T lymphocytes (CTLs) in a one-way mixed lymphocyte reaction. Two unique combinations with a single HLA mismatch in the HLA-B44 serotype differing by one amino acid in the α3 domain were investigated. We show that the B*44:27 versus B*44:02 mismatch was not recognized by CTLs in both directions. At days 10 and 20, the frequency of CD8+CD137+T cells was comparable to that measured in the autologous stimulation (0.3-3.9%). A B*44:02 versus B*44:03 mismatch was, however, well recognized at day 10 (7.2%) and day 20 (17.8%). This is the first demonstration that a single HLA-B mismatch involving a residue outside the peptide-binding site is not recognized in an in vitro functional assay and may probably be considered as a permissive incompatibility in vivo.


Syncytia induction enhances the oncolytic potential of vesicular stomatitis virus in virotherapy for cancer.
Vesicular stomatitis virus (VSV) selectively replicates in tumor but not in normal cells and is being developed as an oncolytic agent for cancer therapy. Here we report the construction of a recombinant VSV capable of inducing syncytia formation between tumor cells through membrane fusion at neutral pH, which led to enhanced oncolytic properties against multifocal hepatocellular carcinoma (HCC) in the livers of immunocompetent rats. Recombinant VSV vectors were constructed by insertion into their genome a transcription unit expressing a control or fusion protein derived from Newcastle disease virus. In vitro characterization of the recombinant fusogenic VSV vector on human and rat HCC cells showed extensive syncytia formation and significantly enhanced cytotoxic effects. In vivo, administration of fusogenic VSV into the hepatic artery of Buffalo rats bearing syngeneic multifocal HCC lesions in their livers resulted in syncytia formation exclusively within the tumors, and there was no collateral damage to the neighboring hepatic parenchyma. The fusogenic VSV also conferred a significant survival advantage over a nonfusogenic control virus in the treated animals (P = 0.0078, log-rank test). The results suggest that fusogenic VSV can be developed into an effective and safe therapeutic agent for cancer treatment in patients, including those with multifocal HCC in the liver.


Is the use of computer navigation in total knee arthroplasty improving implant positioning and function? A comparative study of 198 knees operated at a Norwegian district hospital.
There are few Scandinavian studies on the effect of computer assisted orthopedic surgery (CAOS) in total knee arthroplasty (TKA), compared to conventional technique (CON), and there is little information on effects in pain and function scores. This retrospective study has evaluated the effects of CAOS on radiological parameters and pain, function and quality of life after primary TKA. 198 primary TKAs were operated by one surgeon in two district hospitals; 103 CAOS and 95 CON. The groups were evaluated based on 3 months post-operative radiographs and a questionnaire containing the knee osteoarthritis outcome score (KOOS), the EQ-5D index score and a visual analogue scale (VAS) two years after surgery. Multiple linear regression method was used to investigate possible impact from exposure (CON or CAOS). On hip-knee-ankle radiographs, 20% of measurements were > ±3° of neutral in the CAOS group and 25% in the CON group (p = 0.37). For the femoral component, the number was 5% for CAOS and 18% for CON (p < 0.01). For the tibial component, the difference was not statistically significant (p = 0.58). In the sagittal plane, the surgeon tended to apply more femoral flexion and more posterior tibial slope with CAOS. We observed no statistically or clinically significant difference in KOOS score, VAS or ∆EQ-5D (all p values >0.05), but there was a trend towards better scores for CAOS. Operation time was 3 minutes longer for CON (p = 0.37). CAOS can improve radiological measurements in primary TKA, and makes it possible to adjust component placement to the patient's anatomy. Over-all, the two methods are equal in pain, function and quality-of-life scores.


Comparison of the Head and Neck Positions in Ridden Horses Advertised in an Australian Horse Sales Magazine: 2005 Versus 2018.
The impact of head and neck position (HNP) on horse welfare has received much attention in the scientific literature within the last two decades. Studies have identified physiological and behavioral signs of distress in horses ridden for prolonged time in an HNP with their noseline behind the vertical (BTV), which may compromise their welfare. The objective of this study was to compare potential differences of HNPs shown in horse sales photographs advertised in an Australian horse sales magazine (Horse Deals) from the years 2005 and 2018. In addition, factors potentially impacting HNPs, such as type of tack presented in (e.g., noseband type), riding discipline, and competition experience of the horse, were investigated. The sample population (n = 570) comprised horses ridden with headgear and bit in walk, trot, or canter/gallop, advertised in an Australian horse sales magazine. Issues from April 2005 and October/November 2018 were selected. Head and neck position was categorized as BTV, on the vertical (OV), slightly in front of the vertical (IFV), or extremely in front of the vertical (EIFV; any HNP >30° IFV). Data were analyzed using the chi-squared test and post hoc testing via a multiple regression approach through SPSS and test of proportions via the Z-score calculator for two independent population proportions. Analysis of combined data from years 2005 and 2018 showed 47.0% (n = 570) of the horse sample population were advertised with HNPs BTV. Behind-the-vertical HNP was observed as the predominant HNP (57.8%; n = 268) in the warmblood/eventers/show/performance (WESP) category (P < .0005). In 2005, 53.4% (n = 303) of the sample population were ridden BTV compared with 39.7% (n = 267) in 2018 (P < .001), 12.9% (n = 303) were OV in 2005 compared with 15.0% (n = 267) in 2018 (P > .05), and 10.9% (n = 303) were IFV in 2005 compared with 27.3% (n = 267) in 2018 (P < .0004). These results suggest a positive development with fewer vendors/riders selecting images where the horse's nose was BTV. However, this may be explained by the larger proportion of horses advertised in the WESP category in 2005 (63.0%; n = 303) versus 2018 (28.5%; n = 267), and the WESP category predominantly comprised of dressage, jumper, and eventing horses. In addition, the reduction of HNPs BTV from 53.4% (n = 303) in 2005 to 39.7% (n = 267) in 2018 could be attributed to the observation that in 2018, a larger proportion of horses were listed in categories that do not require the horses to be worked with a flexed HNP referred to as "on-the-bit" (e.g., western, endurance, Australian sStock horses). The HNP BTV remains preferential by a substantial proportion of the horse-owning public when advertising horses for sale, particularly in disciplines where the horse is worked in a flexed HNP or "on-the-bit."


Cellular retinol-binding protein type III is a PPARgamma target gene and plays a role in lipid metabolism.
Cellular retinol-binding protein (CRBP) type III (CRBP-III) belongs to the family of intracellular lipid-binding proteins, which includes the adipocyte-binding protein aP2. In the cytosol, CRBP-III binds retinol, the precursor of retinyl ester and the active metabolite retinoic acid. The goal of the present work is to understand the regulation of CRBP-III expression and its role in lipid metabolism. Using EMSAs, luciferase reporter assays, and chromatin immunoprecipitation assays, we found that CRBP-III is a direct target of peroxisome proliferator-activated receptor-gamma (PPARgamma). Moreover, CRBP-III expression was induced in adipose tissue of mice after treatment with the PPARgamma agonist rosiglitazone. To examine a potential role of CRBP-III in regulating lipid metabolism in vivo, CRBP-III-deficient (C-III-KO) mice were maintained on a high-fat diet (HFD). Hepatic steatosis was decreased in HFD-fed C-III-KO compared with HFD-fed wild-type mice. These differences were partly explained by decreased serum free fatty acid levels and decreased free fatty acid efflux from adipose tissue of C-III-KO mice. In addition, the lack of CRBP-III was associated with reduced food intake, increased respiratory energy ratio, and altered body composition, with decreased adiposity and increased lean body mass. Furthermore, expression of genes involved in mitochondrial fatty acid oxidation in brown adipose tissue was increased in C-III-KO mice, and C-III-KO mice were more cold tolerant than wild-type mice fed an HFD. In summary, we demonstrate that CRBP-III is a PPARgamma target gene and plays a role in lipid and whole body energy metabolism.


The pathogen recognition sensor, NOD2, is variably expressed in patients with pulmonary tuberculosis.
NOD2, an intracellular pathogen recognition sensor, modulates innate defences to muropeptides derived from various bacterial species, including Mycobacterium tuberculosis (MTB). Experimentally, NOD2 attenuates two key putative mycobactericidal mechanisms. TNF-alpha synthesis is markedly reduced in MTB-antigen stimulated-mononuclear cells expressing mutant NOD2 proteins. NOD2 agonists also induce resistance to apoptosis, and may thus facilitate the survival of MTB in infected macrophages. To further define a role for NOD2 in disease pathogenesis, we analysed NOD2 transcriptional responses in pulmonary leucocytes and mononuclear cells harvested from patients with pulmonary tuberculosis (PTB). We analysed NOD2 mRNA expression by real-time polymerase chain-reaction in alveolar lavage cells obtained from 15 patients with pulmonary tuberculosis and their matched controls. We compared NOD2 transcriptional responses, in peripheral leucocytes, before and after anti-tuberculous treatment in 10 patients. In vitro, we measured NOD2 mRNA levels in MTB-antigen stimulated-mononuclear cells. No significant differences in NOD2 transcriptional responses were detected in patients and controls. In some patients, however, NOD2 expression was markedly increased and correlated with toll-like-receptor 2 and 4 expression. In whole blood, NOD2 mRNA levels increased significantly after completion of anti-tuberculosis treatment. NOD2 expression levels did not change significantly in mononuclear cells stimulated with mycobacterial antigens in vitro. There are no characteristic NOD2 transcriptional responses in PTB. Nonetheless, the increased levels of NOD2 expression in some patients with severe tuberculosis, and the increases in expression levels within peripheral leucocytes following treatment merit further studies in selected patient and control populations.


Direct evaluation of reflector effects on radiant flux from InGaN-based light-emitting diodes.
A metal layer formed on the backside of InGaN/sapphire-based light-emitting diodes deteriorates the inherent optical power output. An experimental approach of a suspended die is employed to study the effects of such metal layers via a direct comparison in radiant flux from a discrete die with and without a reflector. A sphere package that employs no reflector is proposed and fabricated. Light extraction of the sphere design is discussed; a light source in the sphere package would not have to be either an ideal point or placed at the center of the sphere, due to a finite critical angle at the sphere/air interface.


Treatment-related Decisional Conflict, Quality of Life, and Comorbidity in Older Adults with Cancer.
The present study aims to examine the relationships between and among cancer treatment-related decisional conflict, quality of life, and comorbidity in older adults with cancer. A convenience sample of 200 older adults was recruited from outpatient medical oncology and radiation oncology practices in the northeastern United States. A cross-sectional, descriptive, correlational study design was used employing a survey method. Survey instruments included the Decisional Conflict scale (DCS) (with five subscales, including informed, values clarity, support, uncertainty, and effective decision); Self-administered comorbidity questionnaire (SCQ); European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (including five function scales, three symptom scales, a global health/quality of life scale, and six single items); and an investigator-developed demographic form. The mean total DCS score was 22.1 (±12.5). The uncertainty subscale had the highest mean of the subscales (29.2 ± 18.2). The mean score for global health status/quality of life was 44.2 (±20.7). The mean score of the SCQ was low (9.6 ± 4.1). Significant positive relationships were identified between decisional conflict and quality of life (P = 0.009) and quality of life and comorbidity (P = 0.001). Multiple linear regression analysis found statistically significant relationships for total decisional conflict score and the five decisional conflict scale subscales. Results may suggest a relationship between decisional conflict and quality of life, as well as the quality of life and comorbidity. In addition, there are several physical, emotional, and spiritual factors that may positively or negatively impact decisional conflict.


The temporal structure of spoken sentence comprehension in Chinese.
Three experiments with 60 native Chinese speakers were conducted to examine the temporal structure of spoken sentence comprehension in Chinese. In the two sentence-gating experiments, listeners heard sentence fragments of various sizes, either successively or individually, and decided on the agent role of the sentence. In the agent-naming experiment, listeners repeated aloud the sentence agent upon hearing the sentence. Converging evidence was derived from these experiments indicating the usefulness of sentence gating in tapping into the temporal structure of comprehension and in evaluating listeners' comprehension on the basis of partial information. Results from these experiments show that partial sentence information plays a very important role in Chinese sentence comprehension, and that Chinese listeners integrate incomplete and ungrammatical sentence fragments by reference to complete and grammatical models in the language. It is argued that these processing characteristics reflect properties of the Chinese language in which few morphological, grammatical, and syntactic constraints are placed on the relationships between sentence constituents.


The relation between information provision and health-related quality of life, anxiety and depression among cancer survivors: a systematic review.
Providing information that is congruent with patients' needs is an important determinant for patient satisfaction and might also affect health-related quality of life (HRQoL) and anxiety and depression levels of cancer survivors. The authors systematically reviewed the available literature on the relationship between information provision and HRQoL, anxiety and depression. A PubMed literature search for original articles published until February 2010 was carried out. Twenty-five articles, all conducted between 1996 and 2009, which met the predefined inclusion criteria, were subjected to a quality checklist. Satisfied patients, patients with fulfilled information needs, and patients who experience less information barriers, in general have a better HRQoL and less anxiety and depression. Out of eight intervention studies that aimed to improve information provision, only one showed a positive association with better HRQoL. Health care providers must pay more attention to patient-centred information provision. Additional research is needed to make definitive conclusions about information interventions as most results did not reach statistical significance due to methodological constraints. The quick development of the relatively young research field of patient-reported outcomes in cancer survivorship will make it possible to conduct better quality studies in the future.


Chlamydia trachomatis-containing vacuole serves as deubiquitination platform to stabilize Mcl-1 and to interfere with host defense.
Obligate intracellular Chlamydia trachomatis replicate in a membrane-bound vacuole called inclusion, which serves as a signaling interface with the host cell. Here, we show that the chlamydial deubiquitinating enzyme (Cdu) 1 localizes in the inclusion membrane and faces the cytosol with the active deubiquitinating enzyme domain. The structure of this domain revealed high similarity to mammalian deubiquitinases with a unique α-helix close to the substrate-binding pocket. We identified the apoptosis regulator Mcl-1 as a target that interacts with Cdu1 and is stabilized by deubiquitination at the chlamydial inclusion. A chlamydial transposon insertion mutant in the Cdu1-encoding gene exhibited increased Mcl-1 and inclusion ubiquitination and reduced Mcl-1 stabilization. Additionally, inactivation of Cdu1 led to increased sensitivity of C. trachomatis for IFNγ and impaired infection in mice. Thus, the chlamydial inclusion serves as an enriched site for a deubiquitinating activity exerting a function in selective stabilization of host proteins and protection from host defense.


Modeling intra-mosquito dynamics of Zika virus and its dose-dependence confirms the low epidemic potential of Aedes albopictus.
Originating from African forests, Zika virus (ZIKV) has now emerged worldwide in urbanized areas, mainly transmitted by Aedes aegypti mosquitoes. Although Aedes albopictus can transmit ZIKV experimentally and was suspected to be a ZIKV vector in Central Africa, the potential of this species to sustain virus transmission was yet to be uncovered until the end of 2019, when several autochthonous transmissions of the virus vectored by Ae. albopictus occurred in France. Aside from these few locally acquired ZIKV infections, most territories colonized by Ae. albopictus have been spared so far. The risk level of ZIKV emergence in these areas remains however an open question. To assess Ae. albopictus' vector potential for ZIKV and identify key virus outbreak predictors, we built a complete framework using the complementary combination of (i) dose-dependent experimental Ae. albopictus exposure to ZIKV followed by time-dependent assessment of infection and systemic infection rates, (ii) modeling of intra-human ZIKV viremia dynamics, and (iii) in silico epidemiological simulations using an Agent-Based Model. The highest risk of transmission occurred during the pre-symptomatic stage of the disease, at the peak of viremia. At this dose, mosquito infection probability was estimated to be 20%, and 21 days were required to reach the median systemic infection rates. Mosquito population origin, either temperate or tropical, had no impact on infection rates or intra-host virus dynamic. Despite these unfavorable characteristics for transmission, Ae. albopictus was still able to trigger and yield large outbreaks in a simulated environment in the presence of sufficiently high mosquito biting rates. Our results reveal a low but existing epidemic potential of Ae. albopictus for ZIKV, that might explain the absence of large scale ZIKV epidemics so far in territories occupied only by Ae. albopictus. They nevertheless support active surveillance and eradication programs in these territories to maintain the risk of emergence to a low level.


Brominated flame retardants and other polyhalogenated compounds in indoor air and dust from two houses in Japan.
This study analyzed polyhalogenated compounds (PHCs) such as brominated flame retardants (BFRs) in indoor air and dust samples from two modern homes in Japan. Concentrations of polychlorinated biphenyls (PCBs) and 2,4,6-tribromophenol (2,4,6-TBP) in exhaust and indoor air of two houses were detected at 10(2)-10(3)pgm(-3) order, which were well above those in outdoor air. For dust samples, the detected polybrominated diphenyl ether (PBDE) and polybrominated dibenzo-p-dioxins/furan concentrations resembled values found in our past study. Interestingly, compared to PBDE concentrations, two orders of magnitude higher concentration (13000 ng g(-1)) was observed for hexabromocyclododecanes (HBCDs) in a dust sample from one house. Based on the calculation of air/dust partition ratio values (Kad), low Kad values (log Kad <or=0) were obtained for high brominated PBDEs (pentaBDEs - decaBDE), tetrabromobisphenol A (TBBPA), and HBCDs, although indoor air contained higher contents of low chlorinated PCBs, low brominated PBDEs, and 2,4,6-TBP. Attention should be given to exposure to 2,4,6-TBP through inhalation of air as well as dust ingestion. Results of X-ray fluorescence analysis show high bromine concentrations in curtain and roll screen samples possessing a high product loading factor. Furthermore, a preliminary estimation was made of the likely magnitude of inhabitants' exposure to PHCs via inhalation and dust ingestion in the two houses.


Differentiation of human embryonic stem cells to dopaminergic neurons in serum-free suspension culture.
The use of human embryonic stem cells (hESCs) as a source of dopaminergic neurons for Parkinson's disease cell therapy will require the development of simple and reliable cell differentiation protocols. The use of cell cocultures, added extracellular signaling factors, or transgenic approaches to drive hESC differentiation could lead to additional regulatory as well as cell production delays for these therapies. Because the neuronal cell lineage seems to require limited or no signaling for its formation, we tested the ability of hESCs to differentiate to form dopamine-producing neurons in a simple serum-free suspension culture system. BG01 and BG03 hESCs were differentiated as suspension aggregates, and neural progenitors and neurons were detectable after 2-4 weeks. Plated neurons responded appropriately to electrophysiological cues. This differentiation was inhibited by early exposure to bone morphogenic protein (BMP)-4, but a pulse of BMP-4 from days 5 to 9 caused induction of peripheral neuronal differentiation. Real-time polymerase chain reaction and whole-mount immunocytochemistry demonstrated the expression of multiple markers of the midbrain dopaminergic phenotype in serum-free differentiations. Neurons expressing tyrosine hydroxylase (TH) were killed by 6-hydroxydopamine (6-OHDA), a neurotoxic catecholamine. Upon plating, these cells released dopamine and other catecholamines in response to K+ depolarization. Surviving TH+ neurons, derived from the cells differentiated in serum-free suspension cultures, were detected 8 weeks after transplantation into 6-OHDA-lesioned rat brains. This work suggests that hESCs can differentiate in simple serum-free suspension cultures to produce the large number of cells required for transplantation studies.


Interpersonal relationship challenges among stroke survivors and family caregivers.
A strong interpersonal relationship after stroke is important for the well-being of survivors and family caregivers. However, as many as 54% of families experience relationship problems after stroke and as many as 38% of couples experience overt conflict. The purpose of this study is to enhance understanding about relationship challenges among stroke dyads and to identify implications for direct practice in social work. Semi-structured interviews were conducted with N = 19 care dyads. Qualitative data were analyzed through an interpretive description lens. Seven themes about relationship challenges were identified. Findings highlight areas to consider in promoting strong relationships between survivors and family caregivers. Social workers may have the opportunity to assist dyads with disrupting negative communication cycles, strengthening empathy and collaboration, and achieving a balance so that each person's needs are met.


Site-specific deformable imaging registration algorithm selection using patient-based simulated deformations.
The accuracy of deformable image registration could have a significant dosimetric impact in radiation treatment planning. Various image registration algorithms have been developed for clinical application. However, validation of these algorithms in the current clinical setting remains subjective, relying on visual assessment and lacking a comparison to the ground-truth deformation. In this study, the authors propose a framework to quantitatively validate various image registration solutions by using patient-based synthetic quality assurance (QA) phantoms, which can be applied on a site-by-site basis. The computer-simulated deformation was first generated with virtual deformation QA software and further benchmarked using a physical pelvic phantom that was modeled after real patient CT images. After the validity of the virtual deformation was confirmed, a set of synthetic deformable images was produced to simulate various anatomical movements during radiotherapy based on real patient CT images. Three patients with head-and-neck, prostate, and spine cancer were included. The transformations included bladder filling, soft tissue deformation, mandible, and vertebral body movement, etc., which provided various ground-truth images to validate deformable registration. Several clinically available deformable registration algorithms were tested on these images with multiple registration setups, such as global or regional and single-pass or multipass optimization. The generated deformation fields and the ground-truth deformation are compared using voxel-by-voxel based analysis as well as regional based analysis. Performance of registration algorithms varies with clinical sites. The voxel-by-voxel analysis showed the intensity-based free-form deformation by MIM generated the greatest accuracy for low-contrast small regions that underwent significant deformation, such as bladder expansion for prostate. However, for large field deformations with strong contrast, this approach may increase errors, which is especially evident in the cranial spinal irradiation (CSI) case. Both single-pass and multipass B-spline registrations performed well for the head-and-neck patient and CSI patients. QA for deformable image registration is essential to verify the cumulated dose for accurate adaptive radiotherapy. In this study, the authors develop a workflow that can validate image registration techniques for several different clinical sites and for various types of deformations using patient-based simulated deformations. This work could provide a reference for clinicians and radiation physicists on how to choose appropriate image registration algorithms for different situations.


Measurement of rumen dry matter and neutral detergent fiber degradability of feeds by Fourier-transform infrared spectroscopy.
This study explored the potential of partial least squares (PLS) and Fourier-transform infrared spectroscopy (FTIR) to predict rumen dry matter (DM) and neutral detergent fiber (NDF) degradation parameters of a wide range of feeds for ruminants, as an alternative to the in situ method. In total, 663 samples comprising 80 different feed types were analyzed. In situ DM and NDF degradabilities were determined as follows: effective degradability (ED), rumen soluble fraction (A), degradable but not soluble fraction (B), rate of degradation of the B fraction (C), and indigestible NDF (iNDF). Infrared spectra of dry samples were collected by attenuated total reflectance from 600 to 4000cm(-1). Feeds were randomly classified into 2 subsets of samples with representation of all feed types; one subset was used to develop regression models using partial least squares, and the second subset was used to conduct an external validation of the models. This study indicated that universal models containing all feed types and specific models containing concentrate feeds could provide only a relatively poor estimation of in situ DM degradation parameters because of compositional heterogeneity. More research, such as a particle size distribution analysis, is required to determine whether this lack of accuracy was due to limitations of the FTIR approach, or simply due to methodological error associated with the in situ method. This latter hypothesis may explain the low accuracy observed in the prediction of degradation rates if there was physical leakage of fine particles from the mesh bags used during in situ studies. In contrast, much better predictions were obtained when models were developed for forage feeds alone. Models for forages led to accurate predictions of DMA, DMB, NDFED, and NDF concentration (R(2)=0.91, 0.89, 0.85, and 0.79, standard error = 4.34, 5.97, 4.59, and 4.41% of DM, respectively), and could be used for screening of DMED, NDFC, and iNDF. These models relied on certain regions of the FTIR spectrum (900-1150 and 1500-1700cm(-1)), which are mainly compatible with absorption of plant cell wall components, such as cellulose, pectin, lignin, cutin, and suberin, but also with nonstructural carbohydrates and certain active compounds. In conclusion, FTIR spectroscopy could be considered a low-cost alternative to in situ measurements in feed evaluation.


Tricyclic pyrazoles. 3. Synthesis, biological evaluation, and molecular modeling of analogues of the cannabinoid antagonist 8-chloro-1-(2',4'-dichlorophenyl)-N-piperidin-1-yl-1,4,5,6-tetrahydrobenzo[6,7]cyclohepta[1,2-c]pyrazole-3-carboxamide.
A series of analogues of 8-chloro-1-(2',4'-dichlorophenyl)-N-piperidin-1-yl-1,4,5,6-tetrahydrobenzo[6,7]cyclohepta[1,2-c]pyrazole-3-carboxamide 4a (NESS 0327) (Ruiu, S.; Pinna, G. A.; Marchese, G.; Mussinu, J. M.; Saba, P.; Tambaro, S.; Casti, P.; Vargiu, R.; Pani, L. Synthesis and Characterization of NESS 0327: A Novel Putative Antagonist of CB1 Cannabinoid Receptor. J. Pharmacol. Exp. Ther. 2003, 306, 363-370) was synthesized and evaluated for their affinity to cannabinoid receptors. Depending on the chemical modification of the lead structure that was chosen, compounds 4b, 4c, 4i, 4l, and 4m still proved to be potent binders of the CB1 receptor. Moreover, several analogues (4c, 4d, 4e, and 4m) demonstrated superior CB2 receptor binding affinities compared to the parent ligand. Compounds 4b, 4c, 4i, and 4l displayed the most promising pharmacological profiles, having the highest selectivity for CB1 receptors with Ki(CB2) to Ki(CB1) ratios of 11,250, 2000, 3330 and 4625, respectively. Compound 4c increased the intestinal propulsion in mice and antagonized the effect induced by the CB1 receptor agonist WIN 55,212-2. Finally, molecular modeling studies were carried out on a set of tricyclic pyrazoles (2a-4a) and on rimonabant 1 (SR141716A), indicating that high CB1 receptors affinities were consistent for the tricyclic derivatives, both with a nonplanar geometry of the tricyclic cores and with a precise orientation of the substituent (chlorine) on this ring system.


Collagen organization in the secondary chick cornea during development.
The latter stages of morphogenesis in the embryonic chick cornea are instrumental in the establishment of a properly formed corneal stroma. This study was designed to provide better appreciation of collagen reorganization in the avian corneal stroma during the latter stages of embryogenesis. High-angle synchrotron x-ray diffraction patterns were obtained from 47 developing chick corneas daily at developmental days 13 through 18 (n = 7 or 8 at each time point) and analyzed to establish collagen molecular spacing and fibril orientation. Collagen intermolecular x-ray reflections were of approximately constant intensity between days 13 and 15 of development, but thereafter became progressively more intense, suggesting that extra collagen is deposited in embryonic chick corneas after day 16 of development. At all times, the mean collagen intermolecular spacing measured approximately 1.43 nm. X-ray intensity was not uniform around the intermolecular x-ray reflections at earlier time points. Rather, a fourfold symmetry was evident, indicative of an orthogonal array of collagen fibrils. An index of this symmetry was essentially unchanged between developmental days 13 and 15, but thereafter diminished considerably. The lateral spacing of fibril-forming collagen molecules does not change as the chick cornea develops between days 13 and 18. An orthogonal array of collagen fibrils is present in the corneas of developmental day-13 to -18 chicks, but starting at developmental day 16, additional collagen is deposited in a less well-oriented manner and thus acts to obscure the overall orthogonality, with implications for the biomechanical strength and shape of the cornea.


Treatment for HIV-associated cryptococcal meningitis.
Cryptococcal meningitis is a severe fungal infection that occurs primarily in the setting of advanced immunodeficiency and remains a major cause of HIV-related deaths worldwide. The best induction therapy to reduce mortality from HIV-associated cryptococcal meningitis is unclear, particularly in resource-limited settings where management of drug-related toxicities associated with more potent antifungal drugs is a challenge. To evaluate the best induction therapy to reduce mortality from HIV-associated cryptococcal meningitis; to compare side effect profiles of different therapies. We searched the Cochrane Infectious Diseases Group Specialized Register, CENTRAL, MEDLINE (PubMed), Embase (Ovid), LILACS (BIREME), African Index Medicus, and Index Medicus for the South-East Asia Region (IMSEAR) from 1 January 1980 to 9 July 2018. We also searched the World Health Organization International Clinical Trials Registry Platform (WHO ICTRP), ClinicalTrials.gov, and the ISRCTN registry; and abstracts of select conferences published between 1 July 2014 and 9 July 2018. We included randomized controlled trials that compared antifungal induction therapies used for the first episode of HIV-associated cryptococcal meningitis. Comparisons could include different individual or combination therapies, or the same antifungal therapies with differing durations of induction (less than two weeks or two or more weeks, the latter being the current standard of care). We included data regardless of age, geographical region, or drug dosage. We specified no language restriction. Two review authors independently screened titles and abstracts identified by the search strategy. We obtained the full texts of potentially eligible studies to assess eligibility and extracted data using standardized forms. The main outcomes included mortality at 2 weeks, 10 weeks, and 6 months; mean rate of cerebrospinal fluid fungal clearance in the first two weeks of treatment; and Division of AIDS (DAIDS) grade three or four laboratory events. Using random-effects models we determined pooled risk ratio (RR) and 95% confidence interval (CI) for dichotomous outcomes and mean differences (MD) and 95% CI for continuous outcomes. For the direct comparison of 10-week mortality, we assessed the certainty of the evidence using the GRADE approach. We performed a network meta-analysis using multivariate meta-regression. We modelled treatment differences (RR and 95% CI) and determined treatment rankings for two-week and 10-week mortality outcomes using surface under the cumulative ranking curve (SUCRA). We assessed transitivity by comparing distribution of effect modifiers between studies, local inconsistency through a node-splitting approach, and global inconsistency using design-by-treatment interaction modelling. For the network meta-analysis, we applied a modified GRADE approach for assessing the certainty of the evidence for 10-week mortality. We included 13 eligible studies that enrolled 2426 participants and compared 21 interventions. All studies were carried out in adults, and all but two studies were conducted in resource-limited settings, including 11 of 12 studies with 10-week mortality data.In the direct pairwise comparisons evaluating 10-week mortality, one study from four sub-Saharan African countries contributed data to several key comparisons. At 10 weeks these data showed that those on the regimen of one-week amphotericin B deoxycholate (AmBd) and flucytosine (5FC) followed by fluconazole (FLU) on days 8 to 14 had lower mortality when compared to (i) two weeks of AmBd and 5FC (RR 0.62, 95% CI 0.42 to 0.93; 228 participants, 1 study), (ii) two weeks of AmBd and FLU (RR 0.58, 95% CI 0.39 to 0.86; 227 participants, 1 study), (iii) one week of AmBd with two weeks of FLU (RR 0.49, 95% CI 0.34 to 0.72; 224 participants, 1 study), and (iv) two weeks of 5FC and FLU (RR 0.68, 95% CI 0.47 to 0.99; 338 participants, 1 study). The evidence for each of these comparisons was of moderate certainty. For other outcomes, this shortened one-week AmBd and 5FC regimen had similar fungal clearance (MD 0.05 log10 CFU/mL/day, 95% CI -0.02 to 0.12; 186 participants, 1 study) as well as lower risk of grade three or four anaemia (RR 0.31, 95% CI 0.16 to 0.60; 228 participants, 1 study) compared to the two-week regimen of AmBd and 5FC.For 10-week mortality, the comparison of two weeks of 5FC and FLU with two weeks of AmBd and 5FC (RR 0.92, 95% CI 0.69 to 1.23; 340 participants, 1 study) or two weeks of AmBd and FLU (RR 0.85, 95% CI 0.64 to 1.13; 339 participants, 1 study) did not show a difference in mortality, with moderate-certainty evidence for both comparisons.When two weeks of combination AmBd and 5FC was compared with AmBd alone, pooled data showed lower mortality at 10 weeks (RR 0.66, 95% CI 0.46 to 0.95; 231 participants, 2 studies, moderate-certainty evidence).When two weeks of AmBd and FLU was compared to AmBd alone, there was no difference in 10-week mortality in pooled data (RR 0.94, 95% CI 0.55 to 1.62; 371 participants, 3 studies, low-certainty evidence).One week of AmBd and 5FC followed by FLU on days 8 to 14 was the best induction therapy regimen after comparison with 11 other regimens for 10-week mortality in the network meta-analysis, with an overall SUCRA ranking of 88%. In resource-limited settings, one-week AmBd- and 5FC-based therapy is probably superior to other regimens for treatment of HIV-associated cryptococcal meningitis. An all-oral regimen of two weeks 5FC and FLU may be an alternative in settings where AmBd is unavailable or intravenous therapy cannot be safely administered. We found no mortality benefit of combination two weeks AmBd and FLU compared to AmBd alone. Given the absence of data from studies in children, and limited data from high-income countries, our findings provide limited guidance for treatment in these patients and settings.


Parental age difference, educationally assortative mating and offspring count: evidence from a contemporary population in Taiwan.
Using contemporary population data from Taiwan, we examine the relationships between parental age difference, educationally assortative mating, income and offspring count. Controlling for women's reproductive value (measured by age at first birth), we find that an older husband is associated with fewer offspring, whereas a husband with similar or higher education is associated with more offspring. Concerning resources, we find that women's income is negatively associated with fertility and husband's income is positively associated with fertility among highly educated women. These results are consistent with the view that women compensate for trade-offs between education, income generation and childbearing by seeking mates with a higher status.


Runx1 promotes scar deposition and inhibits myocardial proliferation and survival during zebrafish heart regeneration.
Runx1 is a transcription factor that plays a key role in determining the proliferative and differential state of multiple cell types, during both development and adulthood. Here, we report how Runx1 is specifically upregulated at the injury site during zebrafish heart regeneration, and that absence of runx1 results in increased myocardial survival and proliferation, and overall heart regeneration, accompanied by decreased fibrosis. Using single cell sequencing, we found that the wild-type injury site consists of Runx1-positive endocardial cells and thrombocytes that induce expression of smooth muscle and collagen genes. Both these populations cannot be identified in runx1 mutant wounds that contain less collagen and fibrin. The reduction in fibrin in the mutant is further explained by reduced myofibroblast formation and upregulation of components of the fibrin degradation pathway, including plasminogen receptor annexin 2A as well as downregulation of plasminogen activator inhibitor serpine1 in myocardium and endocardium, resulting in increased levels of plasminogen. Our findings suggest that Runx1 controls the regenerative response of multiple cardiac cell types and that targeting Runx1 is a novel therapeutic strategy for inducing endogenous heart repair.


Distribution of staphylococcal cassette chromosome mec (SCCmec) types I, II, III and IV in coagulase-negative staphylococci from patients attending a tertiary hospital in southern Brazil.
Coagulase-negative staphylococci (CoNS) are now recognized as the aetiological agents of an important range of infections in humans. Most developed countries have reported an increase in CoNS infections in hospitalized patients that are resistant to meticillin and other antibiotics. Staphylococcal cassette chromosome mec (SCCmec) typing is essential for understanding the molecular epidemiology of meticillin-resistant Staphylococcus strains. SCCmec elements are currently classified into types I to VI based on the characteristics of the mec and ccr gene complexes and are further classified into subtypes according to their 'junkyard DNA' region. We evaluated the distribution of SCCmec types in CoNS from patients attending the Hospital de Clínicas de Porto Alegre over the period August 2004-December 2005. Among the 129 bloodstream isolates, 36 (27.9 %) harboured SCCmec type I, 4 (3.0 %) harboured SCCmec type II, 67 (52 %) harboured SCCmec type III, 1 (0.8 %) harboured SCCmec type IV and 4 (3.0 %) harboured SCCmec types I and III. Seventeen isolates were not typable. Identification of CoNS at the species level indicated that Staphylococcus epidermidis was the most common species, with 87 isolates, followed by Staphylococcus haemolyticus (15), Staphylococcus hominis (13), Staphylococcus capitis (12) and Staphylococcus sciuri (1). SCCmec type III was the most prevalent among isolates of S. epidermidis (52 %). Among these strains, 30 (23 %) harboured a modified SCCmec type III which contained an additional dcs region in comparison with regular type III. SCCmec type III was also highly prevalent (75 %) among S. capitis isolates. The predominant SCCmec type found among S. haemolyticus isolates was type I. However, all four isolates harbouring SCCmec type II belonged to S. haemolyticus. Our results indicate that SCCmec type III was the most prevalent among the CoNS. Isolates with SCCmec type III were more resistant to non-beta-lactam antimicrobials than isolates harbouring SCCmec types I, II and IV, although the increase in resistance was statistically significant only for clindamycin (P=0.021), rifampicin (P=0.010) and levofloxacin (P=0.005).


The ecological impacts of marine debris: unraveling the demonstrated evidence from what is perceived.
Anthropogenic debris contaminates marine habitats globally, leading to several perceived ecological impacts. Here, we critically and systematically review the literature regarding impacts of debris from several scientific fields to understand the weight of evidence regarding the ecological impacts of marine debris. We quantified perceived and demonstrated impacts across several levels of biological organization that make up the ecosystem and found 366 perceived threats of debris across all levels. Two hundred and ninety-six of these perceived threats were tested, 83% of which were demonstrated. The majority (82%) of demonstrated impacts were due to plastic, relative to other materials (e.g., metals, glass) and largely (89%) at suborganismal levels (e.g., molecular, cellular, tissue). The remaining impacts, demonstrated at higher levels of organization (i.e., death to individual organisms, changes in assemblages), were largely due to plastic marine debris (> 1 mm; e.g., rope, straws, and fragments). Thus, we show evidence of ecological impacts from marine debris, but conclude that the quantity and quality of research requires improvement to allow the risk of ecological impacts of marine debris to be determined with precision. Still, our systematic review suggests that sufficient evidence exists for decision makers to begin to mitigate problematic plastic debris now, to avoid risk of irreversible harm.


Directly diode-pumped Kerr-lens mode-locked Cr4+:YAG laser.
A directly diode-pumped Kerr-lens mode-locked Cr4+:YAG laser is demonstrated for what is to our knowledge the first time. Pulses as short as 65 fs with up to 30 mW of average output power, at a central wavelength of 1569 nm, were obtained at a repetition rate of 100 MHz. Low-loss chirped mirrors have been used for dispersion compensation up to the third order. Comparison with an Yb-fiber-pumped configuration shows good prospects for improvement.


Calcaneal adduction and eversion are coupled to talus and tibial rotation.
The purpose of this study was to quantify isolated coupling mechanisms of calcaneal adduction/abduction and calcaneal eversion/inversion to proximal bones in vitro. The in vitro approach is necessary because in vivo both movements appear together, making it impossible to determine the extent of their individual contribution to overall ankle joint coupling. Eight fresh frozen foot-leg specimens were tested. Data describing bone orientation and coupling mechanisms between segments were obtained using bone pin marker triads. The bone movement was described in a global coordinate system to examine the coupling between the calcaneus, talus and tibia. The strength of coupling was determined by means of the slope of a linear least squares fit to an angle-angle plot. The coupling coefficients in the present study indicate that not only calcaneal eversion/inversion (coupling coefficient: 0.68 ± 0.15) but to an even greater extent calcaneal adduction/abduction (coupling coefficient: 0.99 ± 0.10) was transferred into talus and tibial rotation, highlighting the relevance of calcaneal adduction for the overall ankle joint coupling. The results of this study present the possibility that controlling calcaneal adduction/abduction can affect talus and tibial rotation and therefore the possible genesis of overuse knee injuries.


A vision physiological estimation of ultraviolet window marking visibility to birds.
Billions of birds are estimated to be killed in window collisions every year, worldwide. A popular solution to this problem may lie in marking the glass with ultraviolet reflective or absorbing patterns, which the birds, but not humans, would see. Elegant as this remedy may seem at first glance, few of its proponents have taken into consideration how stark the contrasts between ultraviolet and human visible light reflections or transmissions must be to be visible to a bird under natural conditions. Complicating matters is that diurnal birds differ strongly in how their photoreceptors absorb ultraviolet and to a lesser degree blue light. We have used a physiological model of avian colour vision to estimate the chromatic contrasts of ultraviolet markings against a natural scene reflected and transmitted by ordinary window glass. Ultraviolets markings may be clearly visible under a range of lighting conditions, but only to birds with a UVS type of ultraviolet vision, such as many passerines. To bird species with the common VS type of vision, ultraviolet markings should only be visible if they produce almost perfect ultraviolet contrasts and are viewed against a scene with low chromatic variation but high ultraviolet content.


Recent advances in forensic anthropology: decomposition research.
Decomposition research is still in its infancy, but significant advances have occurred within forensic anthropology and other disciplines in the past several decades. Decomposition research in forensic anthropology has primarily focused on estimating the postmortem interval (PMI), detecting clandestine remains, and interpreting the context of the scene. Additionally, while much of the work has focused on forensic-related questions, an interdisciplinary focus on the ecology of decomposition has also advanced our knowledge. The purpose of this article is to highlight some of the fundamental shifts that have occurred to advance decomposition research, such as the role of primary extrinsic factors, the application of decomposition research to the detection of clandestine remains and the estimation of the PMI in forensic anthropology casework. Future research in decomposition should focus on the collection of standardized data, the incorporation of ecological and evolutionary theory, more rigorous statistical analyses, examination of extended PMIs, greater emphasis on aquatic decomposition and interdisciplinary or transdisciplinary research, and the use of human cadavers to get forensically reliable data.


Polychaete richness and abundance enhanced in anthropogenically modified estuaries despite high concentrations of toxic contaminants.
Ecological communities are increasingly exposed to multiple chemical and physical stressors, but distinguishing anthropogenic impacts from other environmental drivers remains challenging. Rarely are multiple stressors investigated in replicated studies over large spatial scales (>1000 kms) or supported with manipulations that are necessary to interpret ecological patterns. We measured the composition of sediment infaunal communities in relation to anthropogenic and natural stressors at multiple sites within seven estuaries. We observed increases in the richness and abundance of polychaete worms in heavily modified estuaries with severe metal contamination, but no changes in the diversity or abundance of other taxa. Estuaries in which toxic contaminants were elevated also showed evidence of organic enrichment. We hypothesised that the observed response of polychaetes was not a 'positive' response to toxic contamination or a reduction in biotic competition, but due to high levels of nutrients in heavily modified estuaries driving productivity in the water column and enriching the sediment over large spatial scales. We deployed defaunated field-collected sediments from the surveyed estuaries in a small scale experiment, but observed no effects of sediment characteristics (toxic or enriching). Furthermore, invertebrate recruitment instead reflected the low diversity and abundance observed during field surveys of this relatively 'pristine' estuary. This suggests that differences observed in the survey are not a direct consequence of sediment characteristics (even severe metal contamination) but are related to parameters that covary with estuary modification such as enhanced productivity from nutrient inputs and the diversity of the local species pool. This has implications for the interpretation of diversity measures in large-scale monitoring studies in which the observed patterns may be strongly influenced by many factors that covary with anthropogenic modification.


Topology of transmembrane channel-like gene 1 protein.
Mutations of transmembrane channel-like gene 1 (TMC1) cause hearing loss in humans and mice. TMC1 is the founding member of a family of genes encoding proteins of unknown function that are predicted to contain multiple transmembrane domains. The goal of our study was to define the topology of mouse TMC1 expressed heterologously in tissue culture cells. TMC1 was retained in the endoplasmic reticulum (ER) membrane of five tissue culture cell lines that we tested. We used anti-TMC1 and anti-HA antibodies to probe the topologic orientation of three native epitopes and seven HA epitope tags along full-length TMC1 after selective or complete permeabilization of transfected cells with digitonin or Triton X-100, respectively. TMC1 was present within the ER as an integral membrane protein containing six transmembrane domains and cytosolic N- and C-termini. There is a large cytoplasmic loop, between the fourth and fifth transmembrane domains, with two highly conserved hydrophobic regions that might associate with or penetrate, but do not span, the plasma membrane. Our study is the first to demonstrate that TMC1 is a transmembrane protein. The topologic organization revealed by this study shares some features with that of the shaker-TRP superfamily of ion channels.


The involvement of calcium and MAP kinase signaling pathways in the production of radiation-induced bystander effects.
Much evidence now exists regarding radiation-induced bystander effects, but the mechanisms involved in the transduction of the signal are still unclear. The mitogen-activated protein kinase (MAPK) pathways have been linked to growth factor-mediated regulation of cellular events such as proliferation, senescence, differentiation and apoptosis. Activation of multiple MAPK pathways such as the ERK, JNK and p38 pathways have been shown to occur after exposure of cells to radiation and a variety of other toxic stresses. Previous studies have shown oxidative stress and calcium signaling to be important in radiation-induced bystander effects. The aim of the present study was to investigate MAPK signaling pathways in bystander cells exposed to irradiated cell conditioned medium (ICCM) and the role of oxidative metabolism and calcium signaling in the induction of bystander responses. Human keratinocytes (HPV-G cell line) were irradiated (0.005-5 Gy) using a cobalt-60 teletherapy unit. The medium was harvested 1 h postirradiation and transferred to recipient HPV-G cells. Phosphorylated forms of p38, JNK and ERK were studied by immunofluorescence 30 min-24 h after exposure to ICCM. Inhibitors of the ERK pathway (PD98059 and U0126), the JNK pathway (SP600125), and the p38 pathway (SB203580) were used to investigate whether bystander-induced cell death could be blocked. Cells were also incubated with ICCM in the presence of superoxide dismutase, catalase, EGTA, verapamil, nifedipine and thapsigargin to investigate whether bystander effects could be inhibited because of the known effects on calcium homeostasis. Activated forms of JNK and ERK proteins were observed after exposure to ICCM. Inhibition of the ERK pathway appeared to increase bystander-induced apoptosis, while inhibition of the JNK pathway appeared to decrease apoptosis. In addition, reactive oxygen species, such as superoxide and hydrogen peroxide, and calcium signaling were found to be important modulators of bystander responses. Further investigations of these signaling pathways may aid in the identification of novel therapeutic targets.


Unlocking insights about military children and families.
As this issue of the Future of Children makes clear, we have much yet to learn about military children and their families. A big part of the reason, write Anita Chandra and Andrew London, is that we lack sufficiently robust sources of data. Until we collect more and better data about military families, Chandra and London say, we will not be able to study the breadth of their experiences and sources of resilience, distinguish among subgroups within the diverse military community, or compare military children with their civilian counterparts. After surveying the available sources of data and explaining what they are lacking and why, Chandra and London make several recommendations. First, they say, major longitudinal national surveys, as well as administrative data systems (for example, in health care and in schools), should routinely ask about children's connections to the military, so that military families can be flagged in statistical analyses. Second, questions on national surveys and psychological assessments should be formulated and calibrated for military children to be certain that they resonate with military culture. Third, researchers who study military children should consider adopting a life-course perspective, examining children from birth to adulthood as they and their families move through the transitions of military life and into or out of the civilian world.


Structure and function of the epidermal growth factor domain of P-selectin.
P-selectin is a multidomain adhesion protein on the surface of activated platelets and endothelial cells that functions in the recruitment of leukocytes to the site of inflammation. The amino-terminal lectin and EGF domains constitute the ligand recognition unit. We have produced a synthetic 40-residue P-selectin EGF domain (P-sel:EGF) to examine the structure and function of this domain independent of P-selectin. The peptide was folded in vitro and exhibited the same disulfide bonding pattern as other EGF-like domains. P-sel:EGF did not inhibit P-selectin-mediated cellular adhesion assays, indicating that the lectin domain is also required. We undertook the study of the P-selectin EGF by 1H NMR to determine its structure independent of the lectin domain and to compare its structure to that of E-selectin determined crystallographically [Graves et al. (1994) Nature 367, 532]. Although the binding of P-selectin to its carbohydrate ligand is calcium dependent, and some EGF domains have calcium binding sites, addition of calcium had no effect on the NMR spectrum or on the pH-induced changes. Nearly complete resonance assignments were made from 2D 1H NMR spectra at pH 6.0. Two sections of antiparallel beta-sheet were identified on the basis of the pattern of long-range NOEs, 3JHN alpha coupling constants, and slowly exchanging amides. The solution structure of the peptide backbone was determined using distance geometry and simulated annealing calculations. The backbone RMSD to the geometric average for 19 final structures is 0.64 +/- 0.17 A. The resulting fold closely resembles that of other EGF-like peptides, including the E-selectin EGF domain (RMSD approximately 1.08 A). However, compared to the E-selectin EGF structure which also contains the lectin domain, some residues from 1-11 are less ordered, and novel contacts occur between the amino terminus and the core beta-sheet. Despite marked structural homology of the selectin polypeptide backbones, the selectin EGF surfaces show unique distributions of charged residues, a feature that likely correlates to the functional differences.


Depression, cardiometabolic function and left ventricular hypertrophy in African men and women: the SABPA study.
Depressive symptoms are associated with an increased risk for developing cardiovascular diseases, driven by its link to the metabolic syndrome (MetS). This phenomenon, however, still needs to be investigated in the African population. The aim of this study is to investigate the association between left ventricular hypertrophy (LVH) and MetS risk markers in a determined sample. The researchers stratified Black African men and women into with depressive symptoms (D) or without depressive symptoms (ND) group, based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria score. Fasting MetS, chronic hyperglycemia (HbA1c), ambulatory blood pressure (BP) and Cornell product-LVH (CP-LVH) in ECG measures were obtained. Depressive symptoms were reported in 45.3% of the sample. Independent of depression status, African men and women revealed a pre-diabetic state (glycated hemoglobin >5.7%). CP-LVH was associated with decreased low high-density lipoprotein cholesterol in D African women. In D African men, systolic BP (P = .001) and HbA1c (P = .08) explained 64% and 31% of the variation in LVH, respectively. In conclusion, depressive symptoms in Black African women were associated with a measure of target end organ damage, CP-LVH, and this association was driven by a metabolic factor. In Black African men, independent of depressive symptoms, LVH, was driven by cardiometabolic factors, namely SBP and HbA1c.


A Crowdsourcing Approach to Developing and Assessing Prediction Algorithms for AML Prognosis.
Acute Myeloid Leukemia (AML) is a fatal hematological cancer. The genetic abnormalities underlying AML are extremely heterogeneous among patients, making prognosis and treatment selection very difficult. While clinical proteomics data has the potential to improve prognosis accuracy, thus far, the quantitative means to do so have yet to be developed. Here we report the results and insights gained from the DREAM 9 Acute Myeloid Prediction Outcome Prediction Challenge (AML-OPC), a crowdsourcing effort designed to promote the development of quantitative methods for AML prognosis prediction. We identify the most accurate and robust models in predicting patient response to therapy, remission duration, and overall survival. We further investigate patient response to therapy, a clinically actionable prediction, and find that patients that are classified as resistant to therapy are harder to predict than responsive patients across the 31 models submitted to the challenge. The top two performing models, which held a high sensitivity to these patients, substantially utilized the proteomics data to make predictions. Using these models, we also identify which signaling proteins were useful in predicting patient therapeutic response.


Optical fiber bundle displacement sensor using an ac-modulated light source with subnanometer resolution and low thermal drift.
An optical fiber bundle displacement sensor with subnanometer order resolution and low thermal drift is proposed. The setup is based on a carrier amplifier system and involves techniques to eliminate fluctuation in the light power of the source. The achieved noise level of the sensor was 0.03nm/√Hz. The stability was estimated by comparing the outputs of two different sensors from the same target for 4 ks (67 min). The relative displacements between the fiber bundle ends of the two sensors and the target surface varied in the area of 400 nm depending on the ambient temperature variation at 2 °C. However, the difference in output between the two sensor systems is within 2 nm for more than 1 hour of measurement. It is expected that it would be reduced to within the area of 0.1 nm if the ambient temperature were controlled to within ±0.1 °C. It is concluded that the stability of the sensors is sufficiently good to be used with nanotechnological instruments.


Gating and regulation of connexin 43 (Cx43) hemichannels.
Connexin 43 (Cx43) nonjunctional or "unapposed" hemichannels can open under physiological or pathological conditions. We characterize hemichannels comprised of Cx43 or Cx43-EGFP (Cx43 with enhanced GFP fused to the C terminus) expressed in HeLa cells. Channel opening was induced at potentials greater than +60 mV. Open probability appeared to be very low. No comparable opening was detected in the parental, nontransfected HeLa cells. Conductance of fully open single hemichannels was approximately 220 pS, which is approximately double that of Cx43 cell-cell channels. Cx43 hemichannels exhibited two types of gating: fast transitions (<1 ms) between the fully open state and a substate of approximately 75 pS and slow transitions (>5 ms) between either open state and the fully closed state. Cx43-EGFP hemichannels exhibited only slow transitions (>5 ms) between closed and fully open states. These properties resemble those of the corresponding Cx43 and Cx43-EGFP cell-cell channels. Cx43 with EGFP on the N terminus (EGFP-Cx43) inserted into the surface and formed plaques but did not form hemichannels or cell-cell channels. Hemichannel blockers, 18beta-glycyrrhetinic acid or La3+, blocked depolarization-induced currents. Uptake of ethidium bromide (i) was faster in Cx43 and Cx43-EGFP than parental and EGFP-Cx43 cells, (ii) was directly correlated with Cx43-EGFP expression, (iii) was reduced by hemichannel blockers, and (iv) occurred at the same low rate in EGFP-Cx43 and parental cells. Although hemichannel opening was not detected electrophysiologically at the resting potential, infrequent or brief opening could account for ethidium bromide uptake. Opening of Cx43 hemichannels may mediate normal signaling or be deleterious.