Atypical timing and presentation of periventricular haemorrhagic infarction in preterm infants: the role of thrombophilia.
Periventricular haemorrhagic infarction (PVHI) is a complication of preterm birth associated with cardiorespiratory instability. To date, the role of thrombophilia as a possible additional risk factor in infants with atypical timing and presentation of PVHI has not been investigated. This was a retrospective cohort study of preterm infants who developed PVHI with an atypical timing and presentation either of antenatal onset or late in the postnatal course in the absence of a preceding sudden deterioration of their clinical condition. In infants with atypical PVHI mutation analysis of the factor V Leiden (G1691A), prothrombin (G20210A) gene, and C677T and A1298C polymorphisms in the MTHFR gene was performed, and plasma lipoprotein(a) and homocysteine levels were measured. Sixty-two preterm infants who presented with a PVHI were studied. Seventeen had an atypical presentation (seven males, 10 females; median birthweight 1170g [range 580-1990g]; median gestational age 30.6wks [range 28.7-33.7wks]). The typical PVHI group comprised 28 males and 17 females (median birthweight 1200g [range 670-2210g]; median gestational age 29.6wks [range 25.3-33.6wks]). Among the 17 infants with atypical presentation, the factor V Leiden mutation was found in seven infants (41%) as well as in the mothers of six of these seven infants; in one infant this was concomitant with a prothrombin gene mutation. A polymorphism in the MTHFR gene was also present in these infants. In two infants with an atypical presentation who were tested for this, a mutation in the COL4A1 gene was found (reported previously). All but two of the infants with an atypical presentation developed spastic unilateral cerebral palsy. An atypical presentation of PVHI in preterm infants tends to occur more often in the presence of thrombophilia. Testing of thrombophilia, especially factor V Leiden and prothrombin gene mutation, is recommended in these infants.


Identifying children with anxiety disorders using brief versions of the Spence Children's Anxiety Scale for children, parents, and teachers.
Anxiety disorders are among the most prevalent mental health disorders experienced by children and are associated with significant negative outcomes. Only a minority of affected children, however, access professional help, and a failure to identify children with anxiety disorders presents a key barrier to treatment access. Existing child anxiety questionnaire measures are long and time consuming to complete, limiting their potential for widespread use as identification tools in community settings. We developed a brief questionnaire for parents, children, and teachers using items from the Spence Children's Anxiety Scale (SCAS) and evaluated the new measure's psychometric properties, capacity to discriminate between a community (n = 361) and clinic-referred sample (n = 338) of children aged 7-11, and identified optimal cut-off scores for accurate identification of preadolescent children experiencing clinically significant levels of anxiety. The findings provided support for the reliability and validity of 8-item versions of the SCAS, with the brief questionnaire scores displaying comparable internal consistency, agreement among reporters, and convergent/divergent validity to the full-length SCAS scores. The brief SCAS scores also discriminated between the community and clinic-referred samples and identified children in the clinic-referred sample with a moderate-to-good level of accuracy and acceptable sensitivity and specificity. Combining reporters improved sensitivity, but at the expense of specificity, and findings suggested parent report should be prioritized. This new brief questionnaire has potential for use in community settings as a tool to improve identification of children who are experiencing clinically significant levels of anxiety and warrant further assessment and potential support. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Association of a decreased number of d(CA) repeats in the matrix metalloproteinase-9 promoter with glomerulosclerosis susceptibility in mice.
The genetic background plays an important role in the development of progressive glomerulosclerosis. However, no marker is available for the reliable prediction of genetic susceptibility to glomerulosclerosis. Because matrix metalloproteinase-9 (MMP-9) levels are decreased in models of glomerulosclerosis and MMP-9 promoter polymorphism has been observed among patients with diabetic nephropathy, MMP-9 could be one such marker. The object of this study was to determine whether MMP-9 promoter polymorphism was associated with altered MMP-9 expression in mesangial cells (MC) from two mouse strains, i.e., ROP (glomerulosclerosis prone) and B6SJL (glomerulosclerosis resistant). ROP MC expressed 12-fold less MMP-9 mRNA. The MMP-9 promoter in ROP MC contained fewer d(CA) repeats, which was associated with lower MMP-9 expression and activity. Phorbol-12-myristate-13-acetate (3 to 60 ng/ml) increased MMP-9 expression in both MC types (3- to 4.5-fold), but the level in ROP MC never reached that in B6SLJ MC. Although reciprocal transfection of ROP and B6SJL MMP-9 promoter constructs into B6SJL and ROP cells revealed that the promoters were functional in both cell types, the B6SJL promoter was less responsive to phorbol-12-myristate-13-acetate stimulation when transfected into ROP MC, suggesting a role for other factors. In conclusion, the MMP-9 promoter exhibits a decreased number of d(CA) repeats in the sclerosis-prone strain. Because fewer d(CA) repeats associated with decreased MMP-9 expression in MC, it might be a genetic marker for glomerulosclerosis.


Many-element coupled-resonator optical waveguides using gapless-coupled microdisk resonators.
We report silicon photonic many-element coupled-resonator optical waveguides (CROWs) using microspiral and double-notch-shaped microdisk resonators. Such microresonators enable gapless inter-cavity coupling via seamlessly jointed sub-micrometer-sized notches. Our experiments reveal CROW transmission spectra using up to 101 gaplessly coupled microdisk resonators. We estimate from the transmission measurements an insertion loss of approximately 0.11-approximately 0.24 dB/disk within the transmission bands. We show high-order filtering performance with maximum 3-dB linewidth of approximately 2.5 nm and measurement-limited side-mode suppression ratio of approximately 30 dB. The demonstrated maximum optical delay from a 101-element 0.4-microm-notch CROW is approximately 70 ps at the transmission band center and approximately 110 ps at the sideband.


Development of a molecular cytogenetic recombination assay for barley.
Distal localisation of chiasmata is common to many cereals and grasses, which consigns many genes of the complement to recombination backwaters. Releasing this potential untapped genetic variation for use in advanced breeding programmes is an ambitious and technically demanding challenge, necessitating controlled shifts in the distribution of crossover events. As part of a collaborative programme to manipulate recombination in barley, we are developing a robust and reliable molecular cytogenetic assay for recombination in this species, which will be used to gauge the success of our forward and reverse genetic interventions. Single-locus bacterial artificial chromosome clones and rDNA probes identify the 7 somatic chromosomes of the complement. Meiocytes at pachytene of meiosis were embedded in polyacrylamide and hybridised in situ with centromere and telomere probes, followed by immunolocalisation of the synaptonemal complex-associated protein Asy1 which highlights the bivalents' axes. Optical sectioning, deconvolution and image analysis of the z-stacks of the nuclei allowed the disentanglement of each bivalent and the construction of an accurate meiotic ideogram. The landing of single-locus bacterial artificial chromosomes and the detection of late recombination proteins will complete the assay and provide a means of discerning subtle changes in recombination in this species.


Widespread Polycistronic Transcripts in Fungi Revealed by Single-Molecule mRNA Sequencing.
Genes in prokaryotic genomes are often arranged into clusters and co-transcribed into polycistronic RNAs. Isolated examples of polycistronic RNAs were also reported in some higher eukaryotes but their presence was generally considered rare. Here we developed a long-read sequencing strategy to identify polycistronic transcripts in several mushroom forming fungal species including Plicaturopsis crispa, Phanerochaete chrysosporium, Trametes versicolor, and Gloeophyllum trabeum. We found genome-wide prevalence of polycistronic transcription in these Agaricomycetes, involving up to 8% of the transcribed genes. Unlike polycistronic mRNAs in prokaryotes, these co-transcribed genes are also independently transcribed. We show that polycistronic transcription may interfere with expression of the downstream tandem gene. Further comparative genomic analysis indicates that polycistronic transcription is conserved among a wide range of mushroom forming fungi. In summary, our study revealed, for the first time, the genome prevalence of polycistronic transcription in a phylogenetic range of higher fungi. Furthermore, we systematically show that our long-read sequencing approach and combined bioinformatics pipeline is a generic powerful tool for precise characterization of complex transcriptomes that enables identification of mRNA isoforms not recovered via short-read assembly.


A quasi-monoclonal mouse.
As a model for studying the generation of antibody diversity, a gene-targeted mouse was produced that is hemizygous for a rearranged V(D)J segment at the immunoglobulin (Ig) heavy chain locus, the other allele being nonfunctional. The mouse also has no functional kappa light chain allele. The heavy chain, when paired with any lambda light chain, is specific for the hapten (4-hydroxy-3-nitrophenyl) acetyl (NP). The primary repertoire of this quasi-monoclonal mouse is monospecific, but somatic hypermutation and secondary rearrangements change the specificity of 20 percent of the antigen receptors on B cells. The serum concentrations of the Ig isotypes are similar to those in nontransgenic littermates, but less than half of the serum IgM binds to NP, and none of the other isotypes do. Thus, neither network interactions nor random activation of a small fraction of the B cell population can account for serum Ig concentrations.


Two Delta9-stearic acid desaturases are required for Aspergillus nidulans growth and development.
Unsaturated fatty acids are important constituents of all cell membranes and are required for normal growth. In the filamentous fungus Aspergillus nidulans, unsaturated fatty acids and their derivatives also influence asexual (conidial) and sexual (ascospore) sporulation processes. To investigate the relationship between fatty acid metabolism and fungal development, we disrupted the A. nidulans sdeA and sdeB genes, both encoding Delta9-stearic acid desaturases responsible for the conversion of palmitic acid (16:0) and stearic acid (18:0) to palmitoleic acid (16:1) and oleic acid (18:1). The effects of sdeA deletion on development were profound, such that growth, conidial and ascospore production were all reduced at 22 and 37 degrees C. Total fatty acid content was increased over 3-fold in the DeltasdeA strain, reflected in up-regulation of the expression of the fasA gene encoding the alpha chain of the fatty acid synthase, compared to wild type. Stearic acid accumulated approximately 3-fold compared to wild type in the DeltasdeA strain, while unsaturated fatty acid production was decreased. In contrast, disruption of sdeB reduced fungal growth and conidiation at 22 degrees C, but did not affect these processes at 37 degrees C compared to wild type. Interestingly, ascospore production was increased at 37 degrees C for DeltasdeB compared to wild type. Total fatty acid content was not increased in this strain, although stearic acid accumulated 2-fold compared to wild type, and unsaturated fatty acid production was decreased. Combining the DeltasdeA and DeltasdeB alleles created a synthetic lethal strain requiring the addition of oleic acid to the medium for a modicum of growth. Taken together, our results suggest a role for sdeA in growth and development at all temperatures, while sdeB is involved in growth and development at lower temperatures.


Requirement for 3-phosphoinositide-kependent dinase-1 (PDK-1) in insulin-induced glucose uptake in immortalized brown adipocytes.
To provide insight into the physiological importance of 3-phosphoinositide-dependent kinase-1 (PDK-1) in the metabolic actions of insulin, we have generated mice that harbor a PDK-1 gene containing LoxP sites (PDK-1(lox/lox) mice) and established immortalized brown preadipocyte cell lines both from these animals and from wild-type mice. Exposure to appropriate hormonal inducers resulted in the differentiation of >80% of the immortalized brown preadipocytes derived from both types of mice into mature adipocytes. Introduction of the Cre recombinase with the use of adenovirus-mediated gene transfer induced a dose-dependent decrease in the abundance of PDK-1 in PDK-1(lox/lox) adipocytes but not in the wild-type cells. In Cre-expressing PDK-1(lox/lox) adipocytes in which the abundance of PDK-1 was reduced by approximately 85%, the insulin-induced phosphorylation both of Akt on threonine 308 and of p70 S6 kinase on threonine-389 was markedly inhibited. The phosphorylation both of Akt on serine 473 and of p42 and p44 isoforms of mitogen-activated protein kinase induced by insulin was not affected by Cre expression, indicating that the latter specifically inhibits PDK-1-dependent signaling. Both glucose uptake and the translocation of glucose transporter 4 to the plasma membrane induced by insulin as well as glucose uptake induced by a constitutively active form of phosphoinositide 3-kinase were also greatly inhibited by Cre expression in PDK-1(lox/lox) adipocytes. Phosphorylation of AMP-activated protein kinase and glucose uptake induced by 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) were not affected by Cre expression in PDK-1(lox/lox) adipocytes. These results indicate that PDK-1 is essential for insulin-induced glucose uptake in adipocytes.


Relationship between static posture and rearfoot motion during walking in patellofemoral pain syndrome: effect of a reference posture for gait analysis.
The reference posture used in angular motion calculations may play an important role in the relationship found between static posture and rearfoot motion in the frontal plane in a clinical population such as patients with patellofemoral pain syndrome. This study examined the relationship between rearfoot inversion and eversion during the stance phase of walking and the static relaxed standing measurement in women (aged 18 years and older) with patellofemoral pain syndrome and controls and examined the influence of the reference posture used when calculating dynamic motion. Two reference postures were investigated: vertical alignment between the rearfoot and the lower leg and relaxed calcaneal standing. When using the latter reference posture, a significant correlation was found between the static relaxed standing measurement and peak eversion in controls only. When using the vertical alignment reference posture, significant correlation was found only in the patellofemoral pain syndrome group for peak eversion and inversion. The positive relationship found in the patellofemoral pain syndrome group between dynamic angular measures, based on a neutral reference posture, and static relaxed standing indicated that for subjects with patellofemoral pain syndrome, the clinical rearfoot measurement of relaxed standing can be used to explain the pattern of rearfoot motion during walking.


Cost-effectiveness of intermittent preventive treatment with dihydroartemisinin-piperaquine for malaria during pregnancy: an analysis using efficacy results from Uganda and Kenya, and pooled data.
Prevention of malaria infection during pregnancy in HIV-negative women currently relies on the use of long-lasting insecticidal nets together with intermittent preventive treatment in pregnancy with sulfadoxine-pyrimethamine (IPTp-SP). Increasing sulfadoxine-pyrimethamine resistance in Africa threatens current prevention of malaria during pregnancy. Thus, a replacement for IPTp-SP is urgently needed, especially for locations with high sulfadoxine-pyrimethamine resistance. Dihydroartemisinin-piperaquine is a promising candidate. We aimed to estimate the cost-effectiveness of intermittent preventive treatment in pregnancy with dihydroartemisinin-piperaquine (IPTp-DP) versus IPTp-SP to prevent clinical malaria infection (and its sequelae) during pregnancy. We did a cost-effectiveness analysis using meta-analysis and individual trial results from three clinical trials done in Kenya and Uganda. We calculated disability-adjusted life-years (DALYs) arising from stillbirths, neonatal death, low birthweight, mild and moderate maternal anaemia, and clinical malaria infection, associated with malaria during pregnancy. Cost estimates were obtained from data collected in observational studies, health-facility costings, and from international drug procurement databases. The cost-effectiveness analyses were done from a health-care provider perspective using a decision tree model with a lifetime horizon. Deterministic and probabilistic sensitivity analyses using appropriate parameter ranges and distributions were also done. Results are presented as the incremental cost per DALY averted and the likelihood that an intervention is cost-effective for different cost-effectiveness thresholds. Compared with three doses of sulfadoxine-pyrimethamine, three doses of dihydroartemisinin-piperaquine, delivered to a hypothetical cohort of 1000 pregnant women, averted 892 DALYs (95% credibility interval 274 to 1517) at an incremental cost of US$7051 (2653 to 13 038) generating an incremental cost-effectiveness ratio (ICER) of $8 (2 to 29) per DALY averted. Compared with monthly doses of sulfadoxine-pyrimethamine, monthly doses of dihydroartemisinin-piperaquine averted 534 DALYS (-141 to 1233) at a cost of $13 427 (4994 to 22 895), resulting in an ICER of $25 (-151 to 224) per DALY averted. Both results were highly robust to most or all variations in the deterministic sensitivity analysis. Our findings suggest that among HIV-negative pregnant women with high uptake of long-lasting insecticidal nets, IPTp-DP is cost-effective in areas with high malaria transmission and high sulfadoxine-pyrimethamine resistance. These data provide a comprehensive overview of the current evidence on the cost-effectiveness of IPTp-DP. Nevertheless, before a policy change is advocated, we recommend further research into the effectiveness and costs of different regimens of IPTp-DP in settings with different underlying sulfadoxine-pyrimethamine resistance. Malaria in Pregnancy Consortium, which is funded through a grant from the Bill & Melinda Gates Foundation to the Liverpool School of Hygiene and Tropical Medicine.


Electropherotypes and subgroups of group A rotaviruses circulating among diarrhoeic children in Kano, Nigeria.
It is estimated that about 600,000 children die annually as a result of severe dehydrating diarrhea caused by rotaviruses. The virus is a double stranded RNA (dsRNA) virus with 11 segments. Group A rotaviruses show a characteristic 4-2-3-2 pattern following electrophoresis. The VP6 subgroups, I and II exist. This work was carried out to study the prevalence of rotavirus infection among children 0-5 years with diarrhea in Kano, and to determine the circulating subgroups and electropherotypes and of the rotavirus isolates. Two hundred and eighteen stool specimens from children 0-60 months (198 diarrheic and 20 non-diarrheic) were collected from different hospitals and health care centers in Kano and subjected to group A rotavirus enzyme linked immunosorbent assay (ELISA) to determine presence of group A rotavirus, subgroup ELISA to determine the VP6 subgroups and polyacrylamide gel electrophoresis (PAGE) to determine the electropherotypes present. The long electropherotypes (47.05%) of four variations dominated over the short electropherotype (17.64%). About 11.76% of the isolates were of mixed infection. Dominance of subgroup II (45%) over subgroup I (25%), and the presence of both subgroups I and II (10%) and neither subgroup I nor II (15%) was observed in this study. Information on the genomic diversity of the RNA electropherotypes in this region, Kano, is reported in this study.


Somatic and psychologic predictors of long-term unfavorable outcome after surgical intervention.
To identify somatic and psychologic predictors of pain, functional limitations, global perceived recovery, and quality of life 6 months after surgical intervention. Recent studies have indicated that chronic pain after surgical intervention is more common than previously assumed. Several demographic and somatic predictors of long-term unfavorable outcome have been identified, but little is known about the contribution of psychologic risk factors. A prospective cohort study, including 625 patients undergoing elective surgery at the University Hospital Maastricht, The Netherlands, was conducted between February and August 2003. Psychologic questionnaires were completed preoperatively and acute postoperative pain was recorded until 4 days after the operation. Six months later, all patients received follow-up questionnaires to assess pain, functional limitations, global perceived recovery, and quality of life. Multivariable logistic regression analyses were used to estimate relative risk of poor outcome in terms of pain, functional limitations, and global recovery. Multivariable linear regression analysis was used to assess associations with quality of life at 6 months. The most important somatic predictors of unfavorable outcome were duration of the operation and high levels of acute postoperative pain. Patients reporting high levels of pain 4 days after the operation and patients undergoing an operation of longer than 3 hours were at risk for increased pain, increased functional limitations, poor global recovery, and reported lower levels of quality of life 6 months after the operation. Psychologic variables that influenced long-term outcome were preoperative fear of surgery and optimism. Fear of the long-term consequences of the operation was associated with more pain, poor global recovery, and worse quality of life 6 months later, whereas optimism was associated with better recovery and higher quality of life. This study was the first to identify the joint contribution of somatic and psychologic factors to chronic pain, functional limitations, and quality of life 6 months after surgical interventions. It replicates previous findings that intense acute postoperative pain is a risk factor for long-term adverse outcome and also identified additional risk factors, namely, long duration of the operation, ASA status, and preoperative fear of surgery.


Discovery of intense gamma-ray flashes of atmospheric origin.
Detectors aboard the Compton Gamma Ray Observatory have observed an unexplained terrestrial phenomenon: brief, intense flashes of gamma rays. These flashes must originate in the atmosphere at altitudes above at least 30 kilometers in order to escape atmospheric absorption and reach the orbiting detectors. At least a dozen such events have been detected over the past 2 years. The photon spectra from the events are very hard (peaking in the high-energy portion of the spectrum) and are consistent with bremsstrahlung emission from energetic (million-electron volt) electrons. The most likely origin of these high-energy electrons, although speculative at this time, is a rare type of high-altitude electrical discharge above thunderstorm regions.


Prospective study of patients' refusal of antipsychotic medication under a physician discretion review procedure.
The authors examined the refusal of antipsychotic medications and associated outcomes prospectively in a group of 348 psychiatric patients admitted to three acute inpatient units in a state-operated mental health facility in Virginia where psychiatrists have the discretionary power to administer treatment over patients' objections. Newly admitted patients were administered both a questionnaire to ascertain their attitudes toward admission and the Brief Psychiatric Rating Scale (BPRS). Patients who refused antipsychotic medication were identified, and data were collected on the length of refusal and whether the refusal episode was terminated voluntarily or involuntarily. A group of patients compliant with antipsychotic medication was selected for comparison on outcome measures, including the rate of seclusion and restraint and length of hospitalization. Patients who refused treatment were found to have significantly higher BPRS scores than were patients who complied with antipsychotic treatment and more negative attitudes toward hospitalization and past, present, and future treatment. Refusal episodes were brief, on average 2.8 days, and all patients, who refused treatment were treated. When compared with the compliant patients, patients who refused treatment were more likely to be assaultive, were more likely to require seclusion and restraint, and had longer hospitalizations. Psychiatrists exercised their discretion to promptly treat all patients who refused treatment. Nonetheless, these patients suffered more morbidity than compliant patients. This study suggests that the negative sequelae of in-hospital treatment refusal cannot be eliminated by rapid treatment. The policy implications are discussed.


Cardiovascular hemodynamics and vasopressin blockade in DOCA-salt hypertensive rats.
In conscious rats with near-malignant phases of DOCA-salt (DS) hypertension, hemodynamics were studied with microspheres before and after administration of a vasopressin (VP) vasopressor antagonist in relation to plasma VP levels (pVP). Compared to the controls, the DS rats showed significant elevations in mean arterial pressure (MAP), total vascular resistance (TVR), and pVP, and a flow redistribution from kidney and spleen to skeletal muscles and heart, with increased vascular resistance in almost all organs. The antagonist elicited no significant systemic hemodynamic effects in DS rats as a whole; however, two subgroups, responders vs nonresponders, were identified according to the effects on MAP. In responders with a pVP of 29.2 +/- 2.7 (SE) pg/ml, the antagonist lowered MAP (-24.9 +/- 5.9 mm Hg) and TVR significantly, while in nonresponders with a pVP of 15.2 +/- 3.4 pg/ml, there were no effects. The major antagonist-induced regional responses were increased flow and decreased vascular resistance in skeletal muscles and skin in whole DS rats, and additionally in the gastrointestinal tract, portal organs, and testes in the responders. Significant correlations were observed between pVP, MAP, TVR, and depressor responses to the antagonist only when all data for DS and control rats were pooled. Thus, the systemic hemodynamic effects of VP are important only in responders with exceedingly elevated pVP. VP contributes significantly to the regional hemodynamic abnormalities in skeletal muscles and skin in whole DS rats, and also in several other organs in the responders.


Adult emergency department patients with sickle cell pain crisis: a learning collaborative model to improve analgesic management.
The objectives were to report the baseline (prior to quality improvement interventions) patient and visit characteristics and analgesic management practices for each site participating in an emergency department (ED) sickle cell learning collaborative. A prospective, multisite longitudinal cohort study in the context of a learning-collaborative model was performed in three midwestern EDs. Each site formed a multidisciplinary team charged with improving analgesic management for patients with sickle cell disease (SCD). Each team developed a nurse-initiated analgesic protocol for SCD patients (implemented after a baseline data collection period of 3.5 months at one site and 10 months at the other two sites). All sites prospectively enrolled adults with an acute pain crisis and SCD. All medical records for patients meeting study criteria were reviewed. Demographic, health services, and analgesic management data were abstracted, including ED visit frequency data, ED disposition, arrival and discharge pain score, and name and route of initial analgesic administered. Ten interviews per quarter per site were conducted with patients within 14 days of their ED discharge, and subjects were queried about the highest level of pain acceptable at discharge. The primary outcome variable was the time to initial analgesic administration. Variable data were described as means and standard deviations (SDs) or medians and interquartile ranges (IQR) for nonnormal data. A total of 155 patients met study criteria (median age = 32 years, IQR = 24-40 years) with a total of 701 ED visits. Eighty-six interviews were conducted. Most patients (71.6%) had between one and three visits to the ED during the study period. However, after removing Site 3 from the analysis because of the short data enrollment period (3.5 months), which influenced the mean number of visits for the entire cohort, 52% of patients had between one and three ED visits over 10 months, 21% had four to nine visits, and 27% had between 10 and 67 visits. Fifty-nine percent of patients were discharged home. The median time to initial analgesic for the cohort was 74 minutes (IQR = 48-135 minutes). Differences between choice of analgesic agent and route selected were evident between sites. For the cohort, 680 initial analgesic doses were given (morphine sulfate, 42%; hydromorphone, 46%; meperidine, 4%; morphine sulfate and ibuprofen or ketorolac, 7%) using the following routes: oral (2%), intravenous (67%), subcutaneous (3%), and intramuscular (28%). Patients reported a significantly lower targeted discharge pain score (mean +/- SD = 4.19 +/- 1.18) compared to the actual documented discharge pain score within 45 minutes of discharge (mean +/- SD = 5.77 +/- 2.45; mean difference = 1.58, 95% confidence interval = .723 to 2.44, n = 43). While half of the patients had one to three ED visits during the study period, many patients had more frequent visits. Delays to receiving an initial analgesic were common, and post-ED interviews reveal that sickle cell pain patients are discharged from the ED with higher pain scores than what they perceive as desirable.


Does drinking lead to sex? Daily alcohol-sex behaviors and expectancies among college students.
A within-person multilevel approach was used to model the links between alcohol use and sexual behavior among first-year college students, using up to 14 days of data for each person with occasions (Level 1, N = 2879 days) nested within people (Level 2, N = 218 people; 51.4% male). Between-persons (Level 2) effects were gender, relationship status, person means of alcohol use, and alcohol-sex expectancies for sexual affect and sexual drive. Within-person (Level 1) effects were weekend days, number of drinks consumed, and the interaction between drinks consumed and alcohol-sex expectancies. Independent of average alcohol use, consuming more drinks on a given day was associated with a greater likelihood of oral sex and with experiencing more positive consequences of sex that day. Significant Alcohol Use x Alcohol-Sex Expectancies interactions were found for oral sex and total sex behaviors, indicating that individuals with more positive expectancies were more likely to have sex after drinking. The negative association between drinks and condom use was at a trend level of significance. Results support the potential for promoting sexual health by focusing on cross-behavior expectancies among late adolescents.


Infant mortality in South Africa--distribution, associations and policy implications, 2007: an ecological spatial analysis.
Many sub-Saharan countries are confronted with persistently high levels of infant mortality because of the impact of a range of biological and social determinants. In particular, infant mortality has increased in sub-Saharan Africa in recent decades due to the HIV/AIDS epidemic. The geographic distribution of health problems and their relationship to potential risk factors can be invaluable for cost effective intervention planning. The objective of this paper is to determine and map the spatial nature of infant mortality in South Africa at a sub district level in order to inform policy intervention. In particular, the paper identifies and maps high risk clusters of infant mortality, as well as examines the impact of a range of determinants on infant mortality. A Bayesian approach is used to quantify the spatial risk of infant mortality, as well as significant associations (given spatial correlation between neighbouring areas) between infant mortality and a range of determinants. The most attributable determinants in each sub-district are calculated based on a combination of prevalence and model risk factor coefficient estimates. This integrated small area approach can be adapted and applied in other high burden settings to assist intervention planning and targeting. Infant mortality remains high in South Africa with seemingly little reduction since previous estimates in the early 2000's. Results showed marked geographical differences in infant mortality risk between provinces as well as within provinces as well as significantly higher risk in specific sub-districts and provinces. A number of determinants were found to have a significant adverse influence on infant mortality at the sub-district level. Following multivariable adjustment increasing maternal mortality, antenatal HIV prevalence, previous sibling mortality and male infant gender remained significantly associated with increased infant mortality risk. Of these antenatal HIV sero-prevalence, previous sibling mortality and maternal mortality were found to be the most attributable respectively. This study demonstrates the usefulness of advanced spatial analysis to both quantify excess infant mortality risk at the lowest administrative unit, as well as the use of Bayesian modelling to quantify determinant significance given spatial correlation. The "novel" integration of determinant prevalence at the sub-district and coefficient estimates to estimate attributable fractions further elucidates the "high impact" factors in particular areas and has considerable potential to be applied in other locations. The usefulness of the paper, therefore, not only suggests where to intervene geographically, but also what specific interventions policy makers should prioritize in order to reduce the infant mortality burden in specific administration areas.


Farnesol as an inhibitor and substrate for rabbit liver microsomal P450 enzymes.
Farnesol and the related isoprenoids, geranylgeraniol, geranylgeranyl pyrophosphate, and farnesyl pyrophosphate, are produced in the endoplasmic reticulum of hepatocytes in mammals, and each serve important biological functions. Of these compounds, only farnesol was shown to significantly inhibit rabbit liver microsomal cytochrome P450 enzymes. The observed inhibition appeared to be reversible, and was not strictly competitive, but rather mixed in nature. Of the activities examined, ethoxycoumarin de-ethylase and diclofenac-4-hydroxylase activities were most sensitive to farnesol, with K(I) and K(I)' values between 11 and 40 microM. Caffeine-8-hydroxylation and taxol-6-hydroxylation were not inhibited at all by farnesol. Farnesol appeared to be a P450 substrate, as well as an inhibitor, as indicated by the NADPH-dependent decrease in farnesol concentration in microsomal incubations, and the metabolism was inhibited by CO, which pointed to the involvement of P450 isozymes.


Systematic literature review of the economic burden of spinal muscular atrophy and economic evaluations of treatments.
Spinal muscular atrophy (SMA) is a rare and devastating condition for which new disease-modifying treatments have recently been approved. Given the increasing importance of economic considerations in healthcare decision-making, this review summarizes the studies assessing the cost of SMA and economic evaluations of treatments. A systematic review of the literature in PubMed and Scopus up to 15 September 2020 was conducted according to PRISMA guidelines. Nine studies reporting the annual cost of care of patients with SMA and six evaluations of the cost-effectiveness of SMA treatments were identified. The average annual cost of SMA1, the most frequent and severe form in which symptoms appear before the age of 6 months were similar according to the different studies, ranged from $75,047 to $196,429 per year. The yearly costs for the forms of the later-onset form, called SMA2, SMA3, and SMA4, which were usually pooled in estimates of healthcare costs, were more variable, ranging from $27,157 to $82,474. The evaluations of cost-effectiveness of treatment compared nusinersen treatment against standard of care (n = 3), two treatments (nusinersen and onasemnogene abeparvovec) against each other and no drug treatment (n = 1), nusinersen versus onasemnogene abeparvovec (n = 1), and standard of care versus nusinersen with and without newborn screening (n = 1). The incremental cost-effectiveness ratio (ICER) of nusinersen compared to standard of care in SMA1 ranged from $210,095 to $1,150,455 per quality-adjusted life years (QALY) gained and that for onasemnogene abeparvovec ranged from $32,464 to $251,403. For pre-symptomatic patients, the ICER value ranged from $206,409 to $735,519. The ICERs for later-onset forms of SMA (2, 3 and 4) were more diverse ranging from $275,943 to $8,438,049. This review confirms the substantial cost burden of standard of care for SMA patients and the high cost-effectiveness ratios of the approved drugs at the current price when delivered in post-symptomatic patients. Since few studies have been conducted so far, there is a need for further prospective and independent economic studies in pre- and post-symptomatic patients.


Diaspore trait preferences of dispersing ants.
Elaiosomes of myrmecochorous plant seeds are known to enhance the attraction of diaspore-dispersing ants by serving as a nutritional reward. However, it remained unclear which (nutritional) compounds affect diaspore preferences of ants. We hypothesized that apart from elaiosome/seed-size ratio, volume, and physical surface of diaspores, the quantity and the composition of fatty acids, amino acids, and sugars strongly influence the diaspore preferences of different species. Chemical (nutritional) profiles as well as structural properties of seeds with and without elaiosomes were analyzed and correlated with observed seed choice behavior of ants. Cafeteria experiments in the field confirmed the enhanced attractiveness of elaiosome-bearing seeds for all three ant species tested (Lasius fuliginosus, Myrmica ruginodis, and Temnothorax nylanderi), although seeds lacking elaiosomes also were transported. In multiple-choice cafeteria experiments with simultaneously offered diaspores of 16 plant species with and without elaiosome and with highly varying structural and chemical properties, all three ant species showed distinct preferences for certain diaspore species. Correlation analyses confirmed that the presence of an elaiosome represents the crucial factor that favors ant diaspore dispersal. In addition, the composition and the content of free amino acids, and to varying degrees fatty acids, were found to significantly affect preferences of each ant species, whereas the effect of single fatty acids acting as chemical triggers for diaspore transport by ants, as supposed by several studies, was not confirmed. In conclusion, although at least some diaspore species lacking elaiosomes attract ants for diaspore removal services by presenting nutritional seed coats, the production of elaiosomes seems to provide a worthwhile investment. Elaiosomes ensure rapid diaspore detection and removal due to chemical cue compounds and by offering a highly nutritional food supply, probably fitting the nutritional demands of ants.


Damselflies (Zygoptera) as paratenic hosts for Serpinema trispinosum and its report from turtle hosts from Oklahoma, USA.
Third-stage larvae of the nematode Serpinema trispinosum (Leidy, 1852) were collected from the midgut of four of five species of adult damselflies (Zygoptera) from a non-irrigated restored semipermanent wetland located in Stillwater, Oklahoma, USA. Of the four infected damselfly species, prevalence and mean abundance was highest for the southern spreadwing, Lestes disjunctus australis Walker (10%, 0.2 ± 0.8) and lowest for the familiar bluet, Enallagma civile (Hagen) (2.5%, 0.04 ± 0.3); whereas mean intensities were lowest for the citrine forktail, Ischnura hastata (Say) (1.5 ± 0.5) and the eastern forktail, Ischnura verticalis (Say) (1.0 ± 0). This is the first record of larvae of S. trispinosum from damselflies. Serpinema trispinosum adults have been reported from 18 species of North and Central American freshwater turtles, whereas microcrustaceans such as copepods serve as intermediate hosts and snails, fish and amphibians serve as paratenic hosts in this nematode's life cycle. However, dietary studies of the 18 species of freshwater turtles reported as definitive hosts for S. trispinosum indicate that aquatic insects including damselflies are more commonly reported in turtle diets than are fish or amphibians. Additionally, unlike snails and amphibians, larval damselflies predominantly feed on microcrustaceans, and our observation of S. trispinosum infecting damselflies may reflect the importance of these insects as paratenic hosts. In the present study, we provide new host information and measurements for third-stage larvae of S. trispinosum from damselfly hosts along with measurements for adult male and female S. trispinosum from turtle hosts from Oklahoma, USA.


Safety of long-term large doses of aspartame.
Safety of long-term administration of 75 mg/kg of aspartame per day was evaluated with the use of a randomized, double-blind, placebo-controlled, parallel-group design in 108 male and female volunteers aged 18 to 62 years. Subjects received either aspartame or placebo in capsule form three times daily for 24 weeks. No persistent changes over time were noted in either group in vital signs; body weight; results of standard laboratory tests; fasting blood levels of aspartame's constituent amino acids (aspartic acid and phenylalanine), other amino acids, and methanol; or blood formate levels and 24-hour urinary excretion of formate. There also were no statistically significant differences between groups in the number of subjects experiencing symptoms or in the number of symptoms per subject. These results further document the safety of the long-term consumption of aspartame at doses equivalent to the amount of aspartame in approximately 10 L of beverage per day.


European Association for Neuro-Oncology (EANO) guidelines for palliative care in adults with glioma.
Patients with glioma present with complex palliative care needs throughout their disease trajectory. The life-limiting nature of gliomas and the presence of specific symptoms related to neurological deterioration necessitate an appropriate and early palliative care approach. The multidisciplinary palliative care task force of the European Association of Neuro-Oncology did a systematic review of the available scientific literature to formulate the best possible evidence-based recommendations for the palliative care of adult patients with glioma, with the aim to reduce symptom burden and improve the quality of life of patients and their caregivers, particularly in the end-of-life phase. When recommendations could not be made because of the scarcity of evidence, the task force either used evidence from studies of patients with systemic cancer or formulated expert opinion. Areas of palliative care that currently lack evidence and thus deserve attention for further research are fatigue, disorders of behaviour and mood, interventions for the needs of caregivers, and timing of advance care planning.


The presence of diverse IS elements and an avrPphD homologue that acts as a virulence factor on the pathogenicity plasmid of Erwinia herbicola pv. gypsophilae.
The pathogenicity of Erwinia herbicola pv. gypsophilae (Ehg) and Erwinia herbicola pv. betae (Ehb) is dependent on a native plasmid (pPATH(Ehg) or pPATH(Ehb)) that harbors the hrp gene cluster, genes encoding type III effectors, phytohormones, biosynthetic genes, and several copies of IS1327. Sequence analysis of the hrp-flanking region in pPATH(Ehg) (cosmid pLA150) revealed a cluster of four additional IS elements designated as ISEhel, ISEhe2, ISEhe3, and ISEhe4. Two copies of another IS element (ISEhe5) were identified on the upstream region of the indole-3-acetic acid operon located on the same cosmid. Based on homology of amino acids and genetic organization, ISEhe1 belongs to the IS630 family, ISEhe2 to the IS5 family, ISEhe3 and ISEhe4 to different groups of the IS3 family, and ISEhe5 to the IS1 family. With the exception of ISEhe4, one to three copies of all the other IS elements were identified only in pathogenic strains of Erwinia herbicola pv. gypsophilae and Erwinia herbicola pv. betae whereas ISEhe4 was present in both pathogenic and nonpathogenic strains. An open reading frame that exhibited high identity (89% in amino acids) to AvrPphD of Pseudomonas syringae pv. phaseolicola was present within the cluster of IS elements. An insertional mutation in the AvrPphDEh, reduced gall size in gypsophila by approximately 85%. In addition, remnants of known genes from four different bacteria were detected on the same cosmid.


Concept analysis of collaboration in implementing problem-based learning in nursing education.
The purpose of this concept analysis was to better understand and define collaboration as it relates to the implementation of problem-based learning (PBL). The process of concept analysis was conducted in three phases; namely, theoretical or literature review, empirical or fieldwork; and analysis phases. Rodgers' evolutionary approach was used to clarify the attributes, antecedents, surrogate, related terms and consequences of collaboration in implementing PBL. The search key terms were 'collaboration', 'problem-based learning', 'nursing' and 'nursing education'. The search was performed in the Cumulative Index to Nursing and Allied Health Literature (CINAHL), Medline and PsycINFO databases. The articles were reviewed for trends that would reflect the current knowledge for collaboration as a concept. Descriptive qualitative study was used to collect data purposively from participants of three universities offering PBL in Republic of South Africa and three hospitals where PBL students are placed. Collaboration in implementing PBL is described as using the following terms: interpersonal, interactive and personal process, shared goal and governance. The antecedents of collaboration include commitment and support; common goal; formal agreement; training and development; and monitoring and evaluation of tools and mechanisms. Consequences of collaboration in implementing PBL are as follows: information, resource and expertise sharing; personal development and mentoring; creation of supportive and nurturing environment; professional socialisation; improved students' outcomes; and effective utilisation of resources. Effective collaboration within nursing education and with other healthcare professionals to achieve quality outcomes in an increasingly interdependent higher education system continues to grow in importance.


United on Sunday: The effects of secular rituals on social bonding and affect.
Religious rituals are associated with health benefits, potentially produced via social bonding. It is unknown whether secular rituals similarly increase social bonding. We conducted a field study with individuals who celebrate secular rituals at Sunday Assemblies and compared them with participants attending Christian rituals. We assessed levels of social bonding and affect before and after the rituals. Results showed the increase in social bonding taking place in secular rituals is comparable to religious rituals. We also found that both sets of rituals increased positive affect and decreased negative affect, and that the change in positive affect predicted the change in social bonding observed. Together these results suggest that secular rituals might play a similar role to religious ones in fostering feelings of social connection and boosting positive affect.


Effects of UV filter 4-methylbenzylidene camphor during early development of Solea senegalensis Kaup, 1858.
The inclusion of organic UV filters in personal care products (PCPs) has increased in recent years. 4-Methylbenzylidene camphor (4MBC) is one of the most used UV filters, and thus it is commonly found in aquatic ecosystems, with proved negative effects on aquatic organisms. Effects on early life stages of marine vertebrates are largely unknown. Therefore, the main goal of this work was to evaluate 4MBC effects on Senegalese sole (Solea Senegalensis Kaup, 1858) larvae at different levels of biological organization. S. senegalensis were exposed to increasing concentrations of 4MBC from egg stage until 96 h. Mortality, growth, malformations, behaviour and biochemical responses, including enzymatic biomarkers were studied. The exposure to 4MBC until 96 h post-fertilization (hpf) induced mortality and malformations in a dose-response manner. Besides, reduced growth with increasing concentrations was observed. The exposure to 4MBC also caused alterations on behaviour, including overall lower swimming time during light and dark periods. Biomarker alterations caused by 4MBC included imbalance of neurotransmission related endpoints (increased acetylcholinesterase activity) and decreased activity of enzymes related to anaerobic metabolism (lower cellular lactate dehydrogenase activity) at the lower concentrations tested. Furthermore, our results suggest that 4MBC do not induce oxidative stress in S. senegalensis larvae, since catalase and lipid peroxidation levels were not significantly altered by 4MBC. S. senegalensis revealed to be a good model species for vertebrate animal testing in the marine environment. Sub-lethal concentrations of 4MBC induced toxic effects at all organizational levels. Swimming behaviour was a sensitive endpoint and showed that exposure to 4MBC causes impairment on response to light stimulus which is possibly linked with the observed imbalances on cholinesterase activity in larvae. Conservation concerns along distribution range of S. senegalensis should consider that increasing levels of UV filters in marine environment might have impact on the ecology of the species.


Does operatory field isolation influence the performance of direct adhesive restorations?
To evaluate the influence of different adhesive strategies (etch-and-rinse and self-etching adhesives) and type of field isolation (absolute or relative) on the clinical performance of restorations of noncervical carious lesions (NCCLs). One hundred forty NCCLs were selected from 38 patients, according to previously established inclusion/exclusion criteria, and assigned to one of four groups (n = 35): etch-and-rinse/rubber-dam (ERR), etch-and-rinse/cotton roll (ERC), self-etching/rubber-dam (SER) and self-etching/cotton roll (SEC). The adhesive systems used were: Adper Single Bond 2 (3M ESPE) and Adper SE Plus (3M ESPE), with restorations made using a composite resin (Z350, 3M ESPE). Using the USPHS modified criteria, 140 restorations were evaluated by two calibrated examiners at 5 different times: immediately after placement, at 7 days, and 2, 6, and 12 months. In order to evaluate the presence of gingival recession after the use of the #212 rubber-dam clamp, the clinical crowns of the teeth from groups ERR and SER were measured at six different periods (baseline, immediately, and at 7 days, 2, 6, and 12 months). Data were subjected to McNemar's, chi-square, and Student's t-tests. Both adhesive strategies reduced tooth sensitivity beyond the second period of evaluation (7 days); tooth sensitivity disappeared after the third period of evaluation (2 months). There were no statistically significant differences between the adhesive techniques or isolation techniques, except for a Bravo score for marginal discoloration in group SEC at 6 months, which was significantly different from the other groups. The rubber-dam isolation technique was more uncomfortable for the patient and resulted in short-term gingival recession. No significant differences were found between the types of isolation or adhesive strategy in this clinical evaluation, with the exception of 2 restorations in group SEC that showed marginal discoloration, possibly due to inadequate enamel etching by the self-etching adhesive. Class V restorations perform equally well placed with or without rubber-dam.


InSe: a two-dimensional material with strong interlayer coupling.
Atomically thin, two-dimensional (2D) indium selenide (InSe) has attracted considerable attention due to the large tunability in the band gap (from 1.4 to 2.6 eV) and high carrier mobility. The intriguingly high dependence of the band gap on layer thickness may lead to novel device applications, although its origin remains poorly understood, and is generally attributed to the quantum confinement effect. In this work, we demonstrate via first-principles calculations that strong interlayer coupling may be mainly responsible for this phenomenon, especially in the fewer-layer region, and it could also be an essential factor influencing other material properties of β-InSe and γ-InSe. The existence of strong interlayer coupling manifests itself in three aspects: (i) indirect-to-direct band gap transitions with increasing layer thickness; (ii) fan-like frequency diagrams of the shear and breathing modes of few-layer flakes; and (iii) strong layer-dependent carrier mobilities. Our results indicate that multiple-layer InSe may be deserving of attention from FET-based technologies and may also be an ideal system to study interlayer coupling, possibly inherent in other 2D materials.


Platelets Endocytose Viral Particles and Are Activated via TLR (Toll-Like Receptor) Signaling.
Thrombocytopenia is associated with many viral infections suggesting virions interact with and affect platelets. Consistently, viral particles are seen inside platelets, and platelet activation markers are detected in viremic patients. In this article, we sought mechanistic insights into these virion/platelet interactions by examining how platelets endocytose, traffic, and are activated by a model virion. Approach and Results: Using fluorescently tagged HIV-1 pseudovirions, 3-dimensional structured illumination microscopy, and transgenic mouse models, we probed the interactions between platelets and virions. Mouse platelets used known endocytic machinery, that is, dynamin, VAMP (vesicle-associated membrane protein)-3, and Arf6 (ADP-ribosylation factor 6), to take up and traffic HIV-1 pseudovirions. Endocytosed HIV-1 pseudovirions trafficked through early (Rab4+) and late endosomes (Rab7+), and then to an LC3+ (microtubule-associated protein 1A/1B-light chain 3) compartment. Incubation with virions induced IRAK4 (interleukin 1 receptor-associated kinase 4), Akt (protein kinase B), and IKK (IκB kinase) activation, granule secretion, and platelet-leukocyte aggregate formation. This activation required TLRs (Toll-like receptors) and MyD88 (myeloid differentiation primary response protein 88) but was less extensive and slower than activation with thrombin. In vivo, HIV-1 pseudovirions injection led to virion uptake and platelet activation, as measured by IKK activation, platelet-leukocyte aggregate formation, and mild thrombocytopenia. All were decreased in VAMP-3-/- and, megakaryocyte/platelet-specific, Arf6-/- mice. Similar platelet activation profiles (increased platelet-leukocyte aggregates, plasma platelet factor 4, and phospho-IκBα) were detected in newly diagnosed and antiretroviral therapy-controlled HIV-1+ patients. Collectively, our data provide mechanistic insights into the cell biology of how platelets endocytose and process virions. We propose a mechanism by which platelets sample the circulation and respond to potential pathogens that they take up.


Photographing AIDS: On Capturing a Disease in Pictures of People with AIDS.
The photography of people with AIDS has been subject to numerous critiques in the 1980s and has become a controversial way of visualizing the AIDS epidemic. While most of the scholarly work on AIDS photography is based in cultural studies and concerned with popular representations, the clinical value of photographs of people with AIDS usually remains overlooked. This article addresses photographs as a "way of seeing" AIDS that contributed crucially to the making of the disease entity AIDS within the history of medicine. Cultural studies methods are applied to analyze clinical photography in the case of AIDS, thus contributing to the medical history of AIDS through the lens of photography. The article reveals the conflation of disease morphology and patient identity as a characteristic feature of both clinical photography and a now historical nature of AIDS.


Shared decision making and behavioral impairment: a national study among children with special health care needs.
The Institute of Medicine has prioritized shared decision making (SDM), yet little is known about the impact of SDM over time on behavioral outcomes for children. This study examined the longitudinal association of SDM with behavioral impairment among children with special health care needs (CSHCN). CSHCN aged 5-17 years in the 2002-2006 Medical Expenditure Panel Survey were followed for 2 years. The validated Columbia Impairment Scale measured impairment. SDM was measured with 7 items addressing the 4 components of SDM. The main exposures were (1) the mean level of SDM across the 2 study years and (2) the change in SDM over the 2 years. Using linear regression, we measured the association of SDM and behavioral impairment. Among 2,454 subjects representing 10.2 million CSHCN, SDM increased among 37% of the population, decreased among 36% and remained unchanged among 27%. For CSHCN impaired at baseline, the change in SDM was significant with each 1-point increase in SDM over time associated with a 2-point decrease in impairment (95% CI: 0.5, 3.4), whereas the mean level of SDM was not associated with impairment. In contrast, among those below the impairment threshold, the mean level of SDM was significant with each one point increase in the mean level of SDM associated with a 1.1-point decrease in impairment (0.4, 1.7), but the change was not associated with impairment. Although the change in SDM may be more important for children with behavioral impairment and the mean level over time for those below the impairment threshold, results suggest that both the change in SDM and the mean level may impact behavioral health for CSHCN.


A genetic analysis of intersex, a gene regulating sexual differentiation in Drosophila melanogaster females.
Sex-type in Drosophila melanogaster is controlled by a hierarchically acting set of regulatory genes. At the terminus of this hierarchy lie those regulatory genes responsible for implementing sexual differentiation: genes that control the activity of target loci whose products give rise to sexually dimorphic phenotypes. The genetic analysis of the intersex (ix) gene presented here demonstrates that ix is such a terminally positioned regulatory locus. The ix locus has been localized to the cytogenetic interval between 47E3-6 and 47F11-18. A comparison of the morphological and behavioral phenotypes of homozygotes and hemizygotes for three point mutations at ix indicates that the null phenotype of ix is to transform diplo-X animals into intersexes while leaving haplo-X animals unaffected. Analysis of X-ray induced, mitotic recombination clones lacking ix+ function in the abdomen of diplo-X individuals indicates that the ix+ product functions in a cell-autonomous manner and that it is required at least until the termination of cell division in this tissue. Taken together with previous analyses, our results indicate that the ix+ product is required to function with the female-specific product of doublesex to implement appropriate female sexual differentiation in diplo-X animals.


Persistence, period and precision of autonomous cellular oscillators from the zebrafish segmentation clock.
In vertebrate development, the sequential and rhythmic segmentation of the body axis is regulated by a "segmentation clock". This clock is comprised of a population of coordinated oscillating cells that together produce rhythmic gene expression patterns in the embryo. Whether individual cells autonomously maintain oscillations, or whether oscillations depend on signals from neighboring cells is unknown. Using a transgenic zebrafish reporter line for the cyclic transcription factor Her1, we recorded single tailbud cells in vitro. We demonstrate that individual cells can behave as autonomous cellular oscillators. We described the observed variability in cell behavior using a theory of generic oscillators with correlated noise. Single cells have longer periods and lower precision than the tissue, highlighting the role of collective processes in the segmentation clock. Our work reveals a population of cells from the zebrafish segmentation clock that behave as self-sustained, autonomous oscillators with distinctive noisy dynamics.


Lost working years due to mental disorders: an analysis of the Norwegian disability pension registry.
Mental disorders are prevalent diagnoses in disability benefit statistics, with awards often granted at younger age than for other diagnoses. We aimed to compare the number of lost working years following disability benefit award for mental disorders versus other diagnostic groups. Data from the complete Norwegian official registry over disability benefit incidence, including primary diagnoses, were analyzed for the period 2001 to 2003 (N = 77,067), a time-period without any reform in the disability benefit scheme. Lost working years due to disability benefit award before scheduled age retirement at age 67 were calculated. Musculoskeletal disorders were the commonest reason for disability benefit awards (36.3%) with mental disorders in second place (24.0%). However, mental disorders were responsible for the most working years lost (33.8%) compared with musculoskeletal disorders (29.4%). Individuals awarded disability benefit for a mental disorder were on average 8.9 years younger (46.1 years) than individuals awarded for a musculoskeletal disorder (55.0 years), and 6.9 years younger than individuals awarded for any other somatic disorder (53.0 years). Anxiety and depressive disorders were the largest contributors to lost working years within mental disorders. Age at award is highly relevant when the total burden of different diagnoses on disability benefits is considered. There is great disparity in total number of lost working years due to disability benefit award for different diagnostic groups. The high number of lost working years from mental disorders has serious consequences for both the individual and for the wider society and economy.


Characterizing the use of class rank in evaluating applicants for veterinary internship and residency positions.
To characterize how class rank and other criteria are used to evaluate applicants for veterinary internship and residency positions. Program directors for 572 internship and residency programs. A survey was sent to program directors asking them to score the importance of 7 items (cover letter, letters of reference, curriculum vitae, veterinary class rank, grade point average, grades for classes specifically related to the internship or residency specialty area, and interview) they could use in evaluating applicants for an internship or residency and to rank those 7 items, along with an open item asking participants to list other criteria they used, from most to least important. Responses were obtained for 195 internship and 222 residency programs. For both internship programs and residency programs, mean importance scores assigned to the 7 items resulted in the same ordering from most to least important, with letters of reference, interview, curriculum vitae, and cover letter most important. Rankings of the importance of the 7 items, along with an "other" item, were similar for internship and residency programs; the most important item was a candidate's letters of reference, followed by the interview, cover letter, and curriculum vitae. Results suggested that although most veterinary internship and residency programs consider class rank and overall grade point average when evaluating applicants, these 2 items were not the most important. For both internship and residency programs, the most important items were an applicant's letters of reference, followed by the interview, cover letter, and curriculum vitae. (J Am Vet Med Assoc 2021;258:776-785).


The Relationship Between Violent Behavior in Healthcare Settings and Communication Skills: An Empirical Study on Provincial Hospitals in Ankara.
Violence in the health care is a multifaceted and complex social phenomenon that attracts the attention of researchers and policymakers in many countries around the world. Violence against healthcare workers has several factors, but there are limited data on the association between violence against healthcare workers and the communication skills of healthcare workers. In this study, we examine the relationship between healthcare professionals' communication skills and violent incidents experienced or witnessed by healthcare workers. The sociodemographic and professional data of 296 healthcare professionals were obtained. All participants were assessed using a communication skills inventory. The total and categorical scores of the communication skills inventories were compared between participants who had been exposed to violence and those who had not. It was found that the emotional score of individuals who had suffered more than one physical attack was lower than that of others (P = .037). In addition, nurses were found to have more verbal violent behavior experience than their counterparts. Other comparisons were found to be insignificant. Healthcare professionals should not only evaluate complex problems such as violence in the health care but also emphasize the role of patients and their relatives. Violence against healthcare professionals cannot be based on a simple factor. We believe that there are many factors that lead to violence. The causes of violence in the health care should be examined by future studies.


Co-digestion of press liquids of source-sorted municipal organic waste in anaerobic sludge treatment of municipal wastewater treatment plants.
This paper describes a semi-continuous laboratory-scale investigation of a potential co-substrate for mesophilic anaerobic sludge digestion in a municipal wastewater treatment plant. A feed liquid produced from source-sorted municipal organic waste by pretreatment with a screw press was subjected to the investigation. Quantities produced in press trials as well as the composition of the feed liquid are presented. Mass balances for N, P and chemical oxygen demand are given in order to verify the methane production of the feed liquid in co-digestion with sewage sludge at mesophilic conditions. Hydraulic retention time of the reactors were 14.7 to 16 d and organic loading rates were 1.5 to 2.7 kg volatile solids (VS) per cubic metre per day. The pretreatment by screw press is compared to the production of feed liquids with pulper-based pretreatment processes. While the addition of the feed liquid increased methane production by about 345 ml CH(4)/g VS(in), total solids of the feed liquid were reduced to about 63%. With respect to co-digestion at municipal wastewater treatment plants, several risks associated with the investigated feed liquid are outlined.


Suicide among Arab-Americans.
Arab-American (AA) populations in the US are exposed to discrimination and acculturative stress-two factors that have been associated with higher suicide risk. However, prior work suggests that socially oriented norms and behaviors, which characterize recent immigrant ethnic groups, may be protective against suicide risk. Here we explored suicide rates and their determinants among AAs in Michigan, the state with the largest proportion of AAs in the US. ICD-9/10 underlying cause of death codes were used to identify suicide deaths from among all deaths in Michigan between 1990 and 2007. Data from the 2000 U.S. Census were collected for population denominators. Age-adjusted suicide rates among AAs and non-ethnic whites were calculated by gender using the direct method of standardization. We also stratified by residence inside or outside of Wayne County (WC), the county with the largest AA population in the state. Suicide rates were 25.10 per 100,000 per year among men and 6.40 per 100,000 per year among women in Michigan from 1990 to 2007. AA men had a 51% lower suicide rate and AA women had a 33% lower rate than non-ethnic white men and women, respectively. The suicide rate among AA men in WC was 29% lower than in all other counties, while the rate among AA women in WC was 20% lower than in all other counties. Among non-ethnic whites, the suicide rate in WC was higher compared to all other counties among both men (12%) and women (16%). Suicide rates were higher among non-ethnic white men and women compared to AA men and women in both contexts. Arab ethnicity may protect against suicide in both sexes, but more so among men. Additionally, ethnic density may protect against suicide among Arab-Americans.


Pediatric multidrug-resistant tuberculosis clinical trials: challenges and opportunities.
On June 17, 2016, RESIST-TB, IMPAACT, Vital Strategies, and New Ventures jointly hosted the Pediatric Multidrug Resistant Tuberculosis Clinical Trials Landscape Meeting in Arlington, Virginia, USA. The meeting provided updates on current multidrug-resistant tuberculosis (MDR-TB) trials targeting pediatric populations and adult trials that have included pediatric patients. A series of presentations were given that discussed site capacity needs, community engagement, and additional interventions necessary for clinical trials to improve the treatment of pediatric MDR-TB. This article presents a summary of topics discussed, including the following: current trials ongoing and planned; the global burden of MDR-TB in children; current regimens for MDR-TB treatment in children; pharmacokinetics of second-line anti-tuberculosis medications in children; design, sample size, and statistical considerations for MDR-TB trials in children; selection of study population, design, and treatment arms for a trial of novel pediatric MDR-TB regimens; practical aspects of pediatric MDR-TB treatment trials; and strategies for integrating children into adult tuberculosis trials. These discussions elucidated barriers to pediatric MDR-TB clinical trials and provided insight into necessary next steps for progress in this field. Investigators and funding agencies need to respond to these recommendations so that important studies can be implemented, leading to improved treatment for children with MDR-TB.


Providing a genitourinary medicine colposcopy service.
Between 15 and 30 years ago, the management of women with abnormal cervical cytology fell within the remit of GU Medicine. This involved performing colposcopy. With the introduction of certification for colposcopists in 1998, most GU Medicine clinicians stopped providing a colposcopy service. As certification is not required for using the colposcope to diagnose and manage other ano-genital conditions, a GU Medicine-based colposcopy service was introduced at Addenbrooke's Hospital, Cambridge, UK, to assess young women with post-coital bleeding (PCB). One of the objectives of this study was to review this service. In 2011, local guidelines were implemented advising referral to the department of GU Medicine for women under the age of 40 years presenting with PCB with or without inter-menstrual bleeding (IMB) and with no history of previous cervical pathology. A case note review was undertaken for 357 consecutive patients to document clinical findings and management and to determine whether this was an appropriate route of referral. Cervical pathology was found to be uncommon and easily treated within the GU Medicine setting. The provision of a colposcopy service by GU Medicine nurse practitioners or doctors is achievable but requires appropriate training. Importantly, once obtained, these skills can be easily and usefully transferred to examining the vulva, penis, anus and anal canal.


Identification of microRNA signatures in umbilical cord blood associated with maternal characteristics.
Umbilical cord blood could serve as useful source of blood markers enabling more efficient and reliable prenatal and neonatal diagnostics. MicroRNAs (miRNAs) are ubiquitous in body fluids where they were used for detecting and monitoring various physiological and pathological conditions. In this descriptive study, we aimed to identify changes in miRNA expression profiles associated with basic maternal somatic and epidemiological characteristics. Study is based on 24 mothers from the Pilot phase of CELSPAC: TNG (Central European Longitudinal Studies of Parents and Children: The Next Generation) study. Cord blood was collected at time of delivery and global miRNA profiling was performed using microRNA Ready-to-use PCR Human Panel I+II TaqMan microarrays. Expression profiles were statistically evaluated in relation to maternal age, BMI, pregnancy weight gain, blood type, Rh factor status, allergies during pregnancy, addictive substance abuse and smoking status. We analyzed expression of 752 human mature miRNAs in 24 samples of umbilical cord blood. For all maternal characteristics tested we described a specific signature of significantly deregulated miRNAs (P < 0.05). Analysis revealed seven miRNA associated with maternal age (three increased and four decreased in women younger than 35 years), 14 miRNAs associated with BMI status (five miRNAs increased and nine miRNAs decreased in women with BMI > 25) and nine miRNAs associated with maternal weight gain during pregnancy (eight miRNAs increased, and one miRNA decreased in women with weight gain < 12 kg). Additionally, 17 miRNAs correlated to blood type (two miRNAs decreased in blood type A, 11 increased in blood type B, two miRNAs increased in blood type AB and two miRNAs increased in blood type 0) and 17 miRNAs to Rh status of mother. We also detected seven miRNAs deregulated in umbilical cord blood of women with allergy (four increased and three decreased in women with allergy), four miRNAs associated to addictive substance abuse status (two up- and two downregulated in women with addictive substance abuse) and eight miRNAs associated with maternal cigarette smoking during pregnancy. We successfully described differences in miRNA profiles in umbilical cord blood associated with basic characteristics connected with mother. Our data suggest that miRNAs in umbilical cord blood are detectable and associated with a wide range of maternal characteristics. These results indicate that miRNAs could potentially serve, and should be studied, as biomarkers for screening and diagnosis of pregnancy-associated complications and pathologies.


The community ecology of pathogens: coinfection, coexistence and community composition.
Disease and community ecology share conceptual and theoretical lineages, and there has been a resurgence of interest in strengthening links between these fields. Building on recent syntheses focused on the effects of host community composition on single pathogen systems, we examine pathogen (microparasite) communities using a stochastic metacommunity model as a starting point to bridge community and disease ecology perspectives. Such models incorporate the effects of core community processes, such as ecological drift, selection and dispersal, but have not been extended to incorporate host-pathogen interactions, such as immunosuppression or synergistic mortality, that are central to disease ecology. We use a two-pathogen susceptible-infected (SI) model to fill these gaps in the metacommunity approach; however, SI models can be intractable for examining species-diverse, spatially structured systems. By placing disease into a framework developed for community ecology, our synthesis highlights areas ripe for progress, including a theoretical framework that incorporates host dynamics, spatial structuring and evolutionary processes, as well as the data needed to test the predictions of such a model. Our synthesis points the way for this framework and demonstrates that a deeper understanding of pathogen community dynamics will emerge from approaches working at the interface of disease and community ecology.


Direct detection of illicit drugs from biological fluids by desorption/ionization mass spectrometry with nanoporous silicon microparticles.
Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is a high throughput analytical technique capable of detecting low molecular weight analytes, including illicit drugs, and with potential applications in forensic toxicology as well as athlete and workplace testing, particularly for biological fluids (oral fluids, urine and blood). However, successful detection of illicit drugs using SALDI-MS often requires extraction steps to reduce the inherent complexity of biological fluids. Here, we demonstrate an all-in-one extraction and analytical system consisting of hydrophobically functionalized porous silicon microparticles (pSi-MPs) for affinity SALDI-MS of prescription and illicit drugs. This novel approach allows for the analysis of drugs from multiple biological fluids without sample preparation protocols. The effect of pSi-MP size, pore diameter, pore depth and functionalization on analytical performance is investigated. pSi-MPs were optimized for the rapid and high sensitivity detection of methadone, cocaine and 3,4-methylenedioxymethamphetamine (MDMA). This optimized system allowed extraction and detection of methadone from spiked saliva and clinical urine samples. Furthermore, by detecting oxycodone in additional clinical saliva and plasma samples, we were able to demonstrate the versatility of the pSi-MP SALDI-MS technique.


Inflammatory bowel disease: etiology, pathogenesis and current therapy.
Ulcerative colitis (UC) and Crohn's disease (CD) constitute the two major groups of idiopathic disorders in inflammatory bowel disease (IBD). Environmental factors, genetic factors and immune responses have been considered as the major etiology of IBD. Despite the diversified pathogenesis of the disease, no guaranteed curative therapeutic regimen has been developed so far. This review summarizes the knowledge on the pathophysiology and current treatment approaches of IBD. Since IBD is caused by excessive and tissue- disruptive inflammatory reactions of the gut wall, down-regulation of the immune responses may allow the damaged mucosa to heal and reset the physiological functions of the gut back to normal. Current pharmacotherapy through modulation of neutrophil-derived factors, cytokines, adhesion molecules and reactive oxygen/nitrogen metabolites has been utterly described. Categories of treatment modalities include corticosteroids, aminosalicylates, immunomodulators, antibiotics, probiotics, and a series of unique novel agents. The use of anti-tumor necrosis factor monoclonal antibody (Infliximab), recombinant anti-inflammatory cytokines and related gene therapy has been covered. In addition, discussions on dietary supplementation and heparin treatment are also included. The anti-inflammatory and immunoregulatory potential of investigational agents such as nicotine and the filtered protective compounds from tobacco smoke, as well as active herbal medicinal compounds were tested in our previous experimental works, whereas promising findings have been presented here. With the discovery of novel target-oriented agents, more effective and relatively harmless approaches of IBD therapy could be established to achieve a curative outcome. Indeed, more experimental and clinical studies are needed to confirm the relevance of these therapies.


Child passenger safety laws in the United States, 1978-2010: policy diffusion in the absence of strong federal intervention.
This article examines the diffusion of U.S. state child passenger safety laws, analyzing over-time changes and inter-state differences in all identifiable features of laws that plausibly influence crash-related morbidity and mortality. The observed trend shows many states' continuing efforts to update their laws to be consistent with latest motor vehicle safety recommendations, with each state modifying their laws on average 6 times over the 30-year period. However, there has been a considerable time lag in knowledge diffusion and policy adoption. Even though empirical evidence supporting the protective effect of child restraint devices was available in the early 1970s, laws requiring their use were not adopted by all 50 states until 1986. For laws requiring minors to be seated in rear seats, the first state law adoption did not occur until two decades after the evidence became publicly available. As of 2010, only 12 states explicitly required the use of booster seats, 9 for infant seats and 6 for toddler seats. There is also great variation among states in defining the child population to be covered by the laws, the vehicle operators subject to compliance, and the penalties resulting from non-compliance. Some states cover only up to 4-year-olds while others cover children up to age 17. As of 2010, states have as many as 14 exemptions, such as those for non-residents, non-parents, commercial vehicles, large vehicles, or vehicles without seatbelts. Factors such as the complexity of the state of the science, the changing nature of guidelines (from age to height/weight-related criteria), and the absence of coordinated federal actions are potential explanations for the observed patterns. The resulting uneven policy landscape among states suggests a strong need for improved communication among state legislators, public health researchers, advocates and concerned citizen groups to promote more efficient and effective policymaking.


A brief overview of metal complexes as nuclear imaging agents.
Metallic radionuclides have been instrumental in the field of nuclear imaging for over half a century. While recent years have played witness to a dramatic rise in the use of radiometals as labels for chelator-bearing biomolecules, imaging agents based solely on coordination compounds of radiometals have long played a critical role in the discipline as well. In this work, we seek to provide a brief overview of metal complex-based radiopharmaceuticals for positron emission tomography (PET) and single photon emission computed tomography (SPECT). More specifically, we have focused on imaging agents in which the metal complex itself rather than a pendant biomolecule or targeting moiety is responsible for the in vivo behavior of the tracer. This family of compounds contains metal complexes based on an array of different nuclides as well as probes that have been used for the imaging of a variety of pathologies, including infection, inflammation, cancer, and heart disease. Indeed, two of the defining traits of transition metal complexes-modularity and redox chemistry-have both been creatively leveraged in the development of imaging agents. In light of our audience, particular attention is paid to structure and mechanism, though clinical data is addressed as well. Ultimately, it is our hope that this review will not only educate readers about some of the seminal work performed in this space over the last 30 years but also spur renewed interest in the creation of radiopharmaceuticals based on small metal complexes.


Automatic method of analysis of OCT images in assessing the severity degree of glaucoma and the visual field loss.
In many practical aspects of ophthalmology, it is necessary to assess the severity degree of glaucoma in cases where, for various reasons, it is impossible to perform a visual field test - static perimetry. These are cases in which the visual field test result is not reliable, e.g. advanced AMD (Age-related Macular Degeneration). In these cases, there is a need to determine the severity of glaucoma, mainly on the basis of optic nerve head (ONH) and retinal nerve fibre layer (RNFL) structure. OCT is one of the diagnostic methods capable of analysing changes in both, ONH and RNFL in glaucoma. OCT images of the eye fundus of 55 patients (110 eyes) were obtained from the SOCT Copernicus (Optopol Tech. SA, Zawiercie, Poland). The authors proposed a new method for automatic determination of the RNFL (retinal nerve fibre layer) and other parameters using: mathematical morphology and profiled segmentation based on morphometric information of the eye fundus. A quantitative ratio of the quality of the optic disk and RNFL - BGA (biomorphological glaucoma advancement) was also proposed. The obtained results were compared with the results obtained from a static perimeter. Correlations between the known parameters of the optic disk as well as those suggested by the authors and the results obtained from static perimetry were calculated. The result of correlation with the static perimetry was 0.78 for the existing methods of image analysis and 0.86 for the proposed method. Practical usefulness of the proposed ratio BGA and the impact of the three most important features on the result were assessed. The following results of correlation for the three proposed classes were obtained: cup/disk diameter 0.84, disk diameter 0.97 and the RNFL 1.0. Thus, analysis of the supposed visual field result in the case of glaucoma is possible based only on OCT images of the eye fundus. The calculations and analyses performed with the proposed algorithm and BGA ratio confirm that it is possible to calculate supposed mean defect (MD) of the visual field test based on OCT images of the eye fundus.


Passive smoking by asthmatics: its greater effect on boys than on girls and on older than on younger children.
In 415 nonsmoking asthmatic children who were seen consecutively, asthma symptoms were more severe if the mother was a smoker than if she was a nonsmoker. This applied to both sexes but was more marked in boys than in girls. There were also other indications that sons were the more severely affected: the forced expiratory volume at 1 second, the forced expiratory flow rate during the middle half of the forced vital capacity, and the provocation concentration of histamine needed to result in a 20% decrease in the forced expiratory volume at 1 second were significantly decreased only in the sons, and lung function test results were significantly less in sons than in daughters of mothers who smoked. When the 415 children were stratified according to age, lung function improved significantly with increasing age in the children of nonsmokers; in children of smokers, by contrast, symptoms and lung function test results became progressively worse. As well, there was a correlation between these indications of asthma severity and the number of years the child had been exposed to the mother's smoke. It appeared that, compared with girls, boys were more sensitive to passive smoking, and that its adverse effect increased with age and with duration of exposure.


Identification of genetic loci in Lactobacillus plantarum that modulate the immune response of dendritic cells using comparative genome hybridization.
Probiotics can be used to stimulate or regulate epithelial and immune cells of the intestinal mucosa and generate beneficial mucosal immunomodulatory effects. Beneficial effects of specific strains of probiotics have been established in the treatment and prevention of various intestinal disorders, including allergic diseases and diarrhea. However, the precise molecular mechanisms and the strain-dependent factors involved are poorly understood. In this study, we aimed to identify gene loci in the model probiotic organism Lactobacillus plantarum WCFS1 that modulate the immune response of host dendritic cells. The amounts of IL-10 and IL-12 secreted by dendritic cells (DCs) after stimulation with 42 individual L. plantarum strains were measured and correlated with the strain-specific genomic composition using comparative genome hybridisation and the Random Forest algorithm. This in silico "gene-trait matching" approach led to the identification of eight candidate genes in the L. plantarum genome that might modulate the DC cytokine response to L. plantarum. Six of these genes were involved in bacteriocin production or secretion, one encoded a bile salt hydrolase and one encoded a transcription regulator of which the exact function is unknown. Subsequently, gene deletions mutants were constructed in L. plantarum WCFS1 and compared to the wild-type strain in DC stimulation assays. All three bacteriocin mutants as well as the transcription regulator (lp_2991) had the predicted effect on cytokine production confirming their immunomodulatory effect on the DC response to L. plantarum. Transcriptome analysis and qPCR data showed that transcript level of gtcA3, which is predicted to be involved in glycosylation of cell wall teichoic acids, was substantially increased in the lp_2991 deletion mutant (44 and 29 fold respectively). Comparative genome hybridization led to the identification of gene loci in L. plantarum WCFS1 that modulate the immune response of DCs.


Fruit and vegetable intake and risk of breast cancer by hormone receptor status.
Estrogen receptor-negative (ER(-)) breast cancer has few known or modifiable risk factors. Because ER(-) tumors account for only 15% to 20% of breast cancers, large pooled analyses are necessary to evaluate precisely the suspected inverse association between fruit and vegetable intake and risk of ER(-) breast cancer. Among 993 466 women followed for 11 to 20 years in 20 cohort studies, we documented 19 869 estrogen receptor positive (ER(+)) and 4821 ER(-) breast cancers. We calculated study-specific multivariable relative risks (RRs) and 95% confidence intervals (CIs) using Cox proportional hazards regression analyses and then combined them using a random-effects model. All statistical tests were two-sided. Total fruit and vegetable intake was statistically significantly inversely associated with risk of ER(-) breast cancer but not with risk of breast cancer overall or of ER(+) tumors. The inverse association for ER(-) tumors was observed primarily for vegetable consumption. The pooled relative risks comparing the highest vs lowest quintile of total vegetable consumption were 0.82 (95% CI = 0.74 to 0.90) for ER(-) breast cancer and 1.04 (95% CI = 0.97 to 1.11) for ER(+) breast cancer (P (common-effects) by ER status < .001). Total fruit consumption was non-statistically significantly associated with risk of ER(-) breast cancer (pooled multivariable RR comparing the highest vs lowest quintile = 0.94, 95% CI = 0.85 to 1.04). We observed no association between total fruit and vegetable intake and risk of overall breast cancer. However, vegetable consumption was inversely associated with risk of ER(-) breast cancer in our large pooled analyses.


A comparison of manual neuronal reconstruction from biocytin histology or 2-photon imaging: morphometry and computer modeling.
Accurate 3D reconstruction of neurons is vital for applications linking anatomy and physiology. Reconstructions are typically created using Neurolucida after biocytin histology (BH). An alternative inexpensive and fast method is to use freeware such as Neuromantic to reconstruct from fluorescence imaging (FI) stacks acquired using 2-photon laser-scanning microscopy during physiological recording. We compare these two methods with respect to morphometry, cell classification, and multicompartmental modeling in the NEURON simulation environment. Quantitative morphological analysis of the same cells reconstructed using both methods reveals that whilst biocytin reconstructions facilitate tracing of more distal collaterals, both methods are comparable in representing the overall morphology: automated clustering of reconstructions from both methods successfully separates neocortical basket cells from pyramidal cells but not BH from FI reconstructions. BH reconstructions suffer more from tissue shrinkage and compression artifacts than FI reconstructions do. FI reconstructions, on the other hand, consistently have larger process diameters. Consequently, significant differences in NEURON modeling of excitatory post-synaptic potential (EPSP) forward propagation are seen between the two methods, with FI reconstructions exhibiting smaller depolarizations. Simulated action potential backpropagation (bAP), however, is indistinguishable between reconstructions obtained with the two methods. In our hands, BH reconstructions are necessary for NEURON modeling and detailed morphological tracing, and thus remain state of the art, although they are more labor intensive, more expensive, and suffer from a higher failure rate due to the occasional poor outcome of histological processing. However, for a subset of anatomical applications such as cell type identification, FI reconstructions are superior, because of indistinguishable classification performance with greater ease of use, essentially 100% success rate, and lower cost.


HIV-1 Vpu blocks recycling and biosynthetic transport of the intrinsic immunity factor CD317/tetherin to overcome the virion release restriction.
The intrinsic immunity factor CD317 (BST-2/HM1.24/tetherin) imposes a barrier to HIV-1 release at the cell surface that can be overcome by the viral protein Vpu. Expression of Vpu results in a reduction of CD317 surface levels; however, the mechanism of this Vpu activity and its contribution to the virological antagonism are incompletely understood. Here, we characterized the influence of Vpu on major CD317 trafficking pathways using quantitative antibody-based endocytosis and recycling assays as well as a microinjection/microscopy-based kinetic de novo expression approach. We report that HIV-1 Vpu inhibited both the anterograde transport of newly synthesized CD317 and the recycling of CD317 to the cell surface, while the kinetics of CD317 endocytosis remained unaffected. Vpu trapped trafficking CD317 molecules at the trans-Golgi network, where the two molecules colocalized. The subversion of both CD317 transport pathways was dependent on the highly conserved diserine S52/S56 motif of Vpu; however, it did not require recruitment of the diserine motif interactor and substrate adaptor of the SCF-E3 ubiquitin ligase complex, β-TrCP. Treatment of cells with the malaria drug primaquine resulted in a CD317 trafficking defect that mirrored that induced by Vpu. Importantly, primaquine could functionally replace Vpu as a CD317 antagonist and rescue HIV-1 particle release. HIV efficiently replicates in the human host and induces the life-threatening immunodeficiency AIDS. Mammalian genomes encode proteins such as CD317 that can inhibit viral replication at the cellular level. As a countermeasure, HIV has evolved genes like vpu that can antagonize these intrinsic immunity factors. Investigating the mechanism by which Vpu overcomes the virion release restriction imposed by CD317, we find that Vpu subverts recycling and anterograde trafficking pathways of CD317, resulting in surface levels of the restriction factor insufficient to block HIV-1 spread. This describes a novel mechanism of immune evasion by HIV.


Electrosprayed inulin microparticles for microbiota triggered targeting of colon.
Inulin, a naturally occurring polysaccharide, was acetylated to make it processable by electrospraying, a facile and single step method for microparticle fabrication. Electrospraying process parameters were optimized for fabrication of spherical and monodisperse indomethacin (IDM) loaded inulin acetate (INA) microparticles. The apparent entrapment efficiency of IDM was determined to be 100%, whereas working encapsulation efficiency was estimated to be 35.39 ± 1.63%. Differential scanning calorimetry and X-ray diffraction analysis confirmed molecular dispersion of IDM in an amorphous state within the INA matrix. Finally, the results from in vitro release study performed in simulated gastro-intestinal fluids demonstrated that IDM was released only in simulated colonic fluid that contained inulinase. Therefore, this study demonstrates that acetylation of inulin does not alter its susceptibility to inulinase and that microparticles fabricated from INA can be developed as a colon targeting drug delivery system.


Chemical composition and mosquito repellency of essential oil of Conyza newii propagated in different geographical locations of Kenya.
Previously, essential oil of Conyza newii (Asterale: Asteracea, Oliv. & Hiern) growing in the northern part of West Pokot (35°E, 1°N) of Kenya was shown to be highly repellent [RD50 = 8.9 × 10(-5) mg/cm(2) , 95% confidence interval (CL)] to Anopheles gambiae s.s. Fumigant toxicity of the oil to the mosquito was also demonstrated. The major constituents of the oil were found to be monoterpenoids, including (S)-(-)-perillyl alcohol, (S)-(-)-perillaldehyde, geraniol, (R)-(+)-limonene, trans-β-ocimene and 1,8-cineol. In this study, the chemical composition and repellency of essential oils of the plant seedlings collected from West Pokot (35°E, 1°N) and propagated in seven different geographical regions of Kenya [West Pokot (35°E, 1°N), Kilome (37°E, 1°S), Naivasha (36°E, 0°), Webuye (34°E, 1°N), Nyakach (34°E, 0°), Kericho (35°E, 0°) and Nairobi (36°E, 1°S)] were compared. There were significant variations (P < 0.01, 95% CL) in the relative proportions of the six constituents and this was reflected in the repellency of the essential oils (P < 0.01, 95% CL). Higher repellency of the oil was associated with greater proportions of (S)-(-) perillyl alcohol, (S)-(-)-perillaldehyde and geraniol, and lower repellency was associated with an increased proportion of (R)-(+)-limonene. The results suggest significant epigenetic (chemotypic) variations in the repellency and composition of C. newii essential oils growing in different regions of Kenya.