Change in cardiovascular risk factors following early diagnosis of type 2 diabetes: a cohort analysis of a cluster-randomised trial.
There is little evidence to inform the targeted treatment of individuals found early in the diabetes disease trajectory. To describe cardiovascular disease (CVD) risk profiles and treatment of individual CVD risk factors by modelled CVD risk at diagnosis; changes in treatment, modelled CVD risk, and CVD risk factors in the 5 years following diagnosis; and how these are patterned by socioeconomic status. Cohort analysis of a cluster-randomised trial (ADDITION-Europe) in general practices in Denmark, England, and the Netherlands. A total of 2418 individuals with screen-detected diabetes were divided into quartiles of modelled 10-year CVD risk at diagnosis. Changes in treatment, modelled CVD risk, and CVD risk factors were assessed at 5 years. The largest reductions in risk factors and modelled CVD risk were seen in participants who were in the highest quartile of modelled risk at baseline, suggesting that treatment was offered appropriately. Participants in the lowest quartile of risk at baseline had very similar levels of modelled CVD risk at 5 years and showed the least variation in change in modelled risk. No association was found between socioeconomic status and changes in CVD risk factors, suggesting that treatment was equitable. Diabetes management requires setting of individualised attainable targets. This analysis provides a reference point for patients, clinicians, and policymakers when considering goals for changes in risk factors early in the course of the disease that account for the diverse cardiometabolic profile present in individuals who are newly diagnosed with type 2 diabetes.


Analyses of mitochondrial genes reveal two sympatric but genetically divergent lineages of Rhipicephalus appendiculatus in Kenya.
The ixodid tick Rhipicephalus appendiculatus transmits the apicomplexan protozoan parasite Theileria parva, which causes East coast fever (ECF), the most economically important cattle disease in eastern and southern Africa. Recent analysis of micro- and minisatellite markers showed an absence of geographical and host-associated genetic sub-structuring amongst field populations of R. appendiculatus in Kenya. To assess further the phylogenetic relationships between field and laboratory R. appendiculatus tick isolates, this study examined sequence variations at two mitochondrial genes, cytochrome c oxidase subunit I (COI) and 12S ribosomal RNA (rRNA), and the nuclear encoded ribosomal internal transcribed spacer 2 (ITS2) of the rRNA gene, respectively. The analysis of 332 COI sequences revealed 30 polymorphic sites, which defined 28 haplotypes that were separated into two distinct haplogroups (A and B). Inclusion of previously published haplotypes in our analysis revealed a high degree of phylogenetic complexity never reported before in haplogroup A. Neither haplogroup however, showed any clustering pattern related to either the geographical sampling location, the type of tick sampled (laboratory stocks vs field populations) or the mammalian host species. This finding was supported by the results obtained from the analysis of 12S rDNA sequences. Analysis of molecular variance (AMOVA) indicated that 90.8 % of the total genetic variation was explained by the two haplogroups, providing further support for their genetic divergence. These results were, however, not replicated by the nuclear transcribed ITS2 sequences likely because of recombination between the nuclear genomes maintaining a high level of genetic sequence conservation. COI and 12S rDNA are better markers than ITS2 for studying intraspecific diversity. Based on these genes, two major genetic groups of R. appendiculatus that have gone through a demographic expansion exist in Kenya. The two groups show no phylogeographic structure or correlation with the type of host species from which the ticks were collected, nor to the evolutionary and breeding history of the species. The two lineages may have a wide geographic distribution range in eastern and southern Africa. The findings of this study may have implications for the spread and control of R. appendiculatus, and indirectly, on the transmission dynamics of ECF.


A closed testing procedure to select an appropriate method for updating prediction models.
Prediction models fitted with logistic regression often show poor performance when applied in populations other than the development population. Model updating may improve predictions. Previously suggested methods vary in their extensiveness of updating the model. We aim to define a strategy in selecting an appropriate update method that considers the balance between the amount of evidence for updating in the new patient sample and the danger of overfitting. We consider recalibration in the large (re-estimation of model intercept); recalibration (re-estimation of intercept and slope) and model revision (re-estimation of all coefficients) as update methods. We propose a closed testing procedure that allows the extensiveness of the updating to increase progressively from a minimum (the original model) to a maximum (a completely revised model). The procedure involves multiple testing with maintaining approximately the chosen type I error rate. We illustrate this approach with three clinical examples: patients with prostate cancer, traumatic brain injury and children presenting with fever. The need for updating the prostate cancer model was completely driven by a different model intercept in the update sample (adjustment: 2.58). Separate testing of model revision against the original model showed statistically significant results, but led to overfitting (calibration slope at internal validation = 0.86). The closed testing procedure selected recalibration in the large as update method, without overfitting. The advantage of the closed testing procedure was confirmed by the other two examples. We conclude that the proposed closed testing procedure may be useful in selecting appropriate update methods for previously developed prediction models. Copyright © 2016 John Wiley & Sons, Ltd.


Identification of intrinsic determinants of midbrain dopamine neurons.
The prospect of using cell replacement therapies has raised the key issue of whether elucidation of developmental pathways can facilitate the generation of therapeutically important cell types from stem cells. Here we show that the homeodomain proteins Lmx1a and Msx1 function as determinants of midbrain dopamine neurons, cells that degenerate in patients with Parkinson's disease. Lmx1a is sufficient and required to trigger dopamine cell differentiation. An early activity of Lmx1a is to induce the expression of Msx1, which complements Lmx1a by inducing the proneural protein Ngn2 and neuronal differentiation. Importantly, expression of Lmx1a in embryonic stem cells results in a robust generation of dopamine neurons with a "correct" midbrain identity. These data establish that Lmx1a and Msx1 are critical intrinsic dopamine-neuron determinants in vivo and suggest that they may be essential tools in cell replacement strategies in Parkinson's disease.


Kainic acid on neostriatal neurons intracellularly recorded in vitro: electrophysiological evidence for differential neuronal sensitivity.
The electrophysiological effects produced by different concentrations of kainic acid (KA) were studied by utilizing intracellular recordings from neostriatal slices. In most of the recorded cells (81%), concentrations of KA ranging between 10 and 300 nM produced reversible and dose-dependent membrane depolarizations. Higher concentrations of this agonist caused larger depolarizations and changes of the membrane properties of the recorded neurons not reversible during the time of recording. In a smaller percentage (19%) of the recorded cells, 10-100 nM KA did not produce significant membrane depolarizations; in these neurons, the depolarizations produced by higher concentrations of KA were small and reversible. The 2 populations of neurons showed similar electrophysiological properties and did not reveal differential sensitivity to quisqualic acid (QUIS; 10-30 microM) or to NMDA (10-30 microM). Tetrodotoxin (TTX; 1 microM) did not reduce the depolarizations produced by KA and by NMDA. Low-calcium, cobalt-containing solutions abolished the effects produced by NMDA, but not the KA-induced depolarizations. Kynurenic acid (500 microM) significantly antagonized the depolarizations produced by KA and reduced the changes of the membrane properties caused by high doses of this agonist. In several neurons, KA induced bicuculline-sensitive synaptic depolarizing potentials. Our findings suggest the presence of 2 subpopulations of neostriatal neurons showing differential postsynaptic sensitivity to KA. The differential sensitivity of neostriatal neurons to KA might influence the responses of these cells to glutamatergic cortical inputs and the degenerative changes observed in neostriatal neurons in some pathological conditions.


Uterine cancer and estrogen therapy.
Recent studies concerning the relationship of exogenous estrogens and endometrial cancer have been reviewed, and the objections to their conclusions discussed. Even considering the most serious of these reservations, it is difficult to avoid the tentative conclusion that estrogens are causally linked to endometrial cancer, and that the attributable risk is numerically sizable. Whether the medical costs will be ultimately shown to outweigh the benefits of estrogen replacement therapy will depend largely on whether or not a relationship to breast cancer is shown, and on the magnitude of the effect of estrogens in preventing serious complications of ovarian failure. For the time being, estrogenic treatment of menopausal symptoms should be as brief as possible, should use the lowest dose possible, and should probably be given in cyclic fashion, particularly to women in intact uteruses. Every attempt should be made to give patients a quantitative understanding of both potential risks and benefits before they decide for or against estrogen therapy.


Computed Tomography-Based SYNTAX Score: A Case Report.
The SYNTAX score is an angiographic scoring system that is currently used for evaluating the complexity of coronary lesions. Its efficiency in both long-term prognosis and the selection of treatment modalities has been shown in many studies. However, there is little evidence for the use of this scoring system in CT angiography, which has become a part of coronary artery disease practice. Here, we showed that the SYNTAX score could be used in a feasible way in CT angiography.


Long Non-Coding RNAs: Emerging and Versatile Regulators in Host-Virus Interactions.
Long non-coding RNAs (lncRNAs) are a class of non-protein-coding RNA molecules, which are involved in various biological processes, including chromatin modification, cell differentiation, pre-mRNA transcription and splicing, protein translation, etc. During the last decade, increasing evidence has suggested the involvement of lncRNAs in both immune and antiviral responses as positive or negative regulators. The immunity-associated lncRNAs modulate diverse and multilayered immune checkpoints, including activation or repression of innate immune signaling components, such as interleukin (IL)-8, IL-10, retinoic acid inducible gene I, toll-like receptors 1, 3, and 8, and interferon (IFN) regulatory factor 7, transcriptional regulation of various IFN-stimulated genes, and initiation of the cell apoptosis pathways. Additionally, some virus-encoded lncRNAs facilitate viral replication through individually or synergistically inhibiting the host antiviral responses or regulating multiple steps of the virus life cycle. Moreover, some viruses are reported to hijack host-encoded lncRNAs to establish persistent infections. Based on these amazing discoveries, lncRNAs are an emerging hotspot in host-virus interactions. In this review, we summarized the current findings of the host- or virus-encoded lncRNAs and the underlying mechanisms, discussed their impacts on immune responses and viral replication, and highlighted their critical roles in host-virus interactions.


Exploring the relation of alcohol consumption to risk of breast cancer.
There are lingering questions regarding the relation between alcohol consumption and breast cancer risk in women. The authors performed a meta-analysis of epidemiologic studies carried out through 1999 to examine the dose-response relation and to assess whether effect estimates differed according to various study characteristics. Overall, there was a monotonic increase in the relative risk of breast cancer with alcohol consumption, but the magnitude of the effect was small; in comparison with nondrinkers, women averaging 12 g/day of alcohol consumption (approximately one typical drink) had a relative risk of 1.10 (95% confidence interval (CI): 1.06, 1.14). Estimates of relative risk were 7% greater in hospital-based case-control studies than in cohort studies or community-based case-control studies, 3% greater in studies published before 1990 than in studies published later, and 5% greater in studies conducted outside of the United States than in US studies. The findings of five US cohort studies published since 1990 yielded a relative risk of 1.06 (95% CI: 1.00, 1.11) for consumers of 12 g/day, as compared with nondrinkers. Cohort studies with less than 10 years of follow-up gave estimates 11% higher than cohort studies with longer follow-up periods. No meaningful difference was seen by menopausal status or type of beverage consumed.


Cognitive Predictors of Precautionary Behavior During the COVID-19 Pandemic.
The attempts to mitigate the unprecedented health, economic, and social disruptions caused by the COVID-19 pandemic are largely dependent on establishing compliance to behavioral guidelines and rules that reduce the risk of infection. Here, by conducting an online survey that tested participants' knowledge about the disease and measured demographic, attitudinal, and cognitive variables, we identify predictors of self-reported social distancing and hygiene behavior. To investigate the cognitive processes underlying health-prevention behavior in the pandemic, we co-opted the dual-process model of thinking to measure participants' propensities for automatic and intuitive thinking vs. controlled and reflective thinking. Self-reports of 17 precautionary behaviors, including regular hand washing, social distancing, and wearing a face mask, served as a dependent measure. The results of hierarchical regressions showed that age, risk-taking propensity, and concern about the pandemic predicted adoption of precautionary behavior. Variance in cognitive processes also predicted precautionary behavior: participants with higher scores for controlled thinking (measured with the Cognitive Reflection Test) reported less adherence to specific guidelines, as did respondents with a poor understanding of the infection and transmission mechanism of the COVID-19 virus. The predictive power of this model was comparable to an approach (Theory of Planned Behavior) based on attitudes to health behavior. Given these results, we propose the inclusion of measures of cognitive reflection and mental model variables in predictive models of compliance, and future studies of precautionary behavior to establish how cognitive variables are linked with people's information processing and social norms.


Effect of catch-up growth by various dietary patterns and resveratrol intervention on bone status.
Catch-up growth (CUG) after food restriction can increase the risks for insulin resistance-related diseases, and to our knowledge, no previous studies have addressed how bone is influenced by CUG when refeeding diet content differs. The objective of this study was to investigate the bone status resulting from CUG induced by varying refeeding dietary patterns, and to assess the potential influencing factors and the effect of resveratrol on bone status during CUG. Experimental rats were randomly divided into five groups: normal chow (NC) group; CUG group (CUG, containing two subgroups, respectively, refeeding with normal chow or high-fat diet); high-fat diet (HF) group; and resveratrol intervention groups (CUGE and HFE). Bone parameters were detected by dual-energy X-ray absorptiometry. Serum concentrations of tumor necrosis factor (TNF)-α, body weight and food intake were also recorded. Our results showed that food restriction induced a significant decrease in bone parameters. Eight-week CUG by normal chow had a greater degree of improvement in bone mineral density than high-fat diet, and even returned to normal level similar to NC. Bone parameters were elevated in varying degrees in the HF group compared with the NC group. In the resveratrol intervention groups, bone parameters significantly increased. Furthermore, bone parameters were inversely related with serum TNF-α concentrations, but showed positive correlation with body weight. In conclusion, the study shows that CUG can partially reverse the deleterious effects of caloric restriction on bone health, especially in the refeeding with normal chow group. Moreover, resveratrol has a protective effect on bone status during the period of CUG. Serum TNF-α levels and body weight also seem to play an important role in regulating bone parameters.


Quality Decision Making in Health Technology Assessment: Issues Facing Companies and Agencies.
To evaluate the quality of the decision-making processes of pharmaceutical companies during medicines development for evidence generation to support reimbursement of new medicines and the appraisal recommendation decision-making process by health technology assessment (HTA) agencies. Two questionnaires were developed and subsequently piloted for the purpose of content validation. These were sent to 24 pharmaceutical companies and 16 HTA agencies. Responses were obtained from 11 companies and 11 HTA agencies. Some similarities were identified between the decision-making processes of companies and agencies, such as the use of committees, having a primarily mixed (qualitative/quantitative) internal decisionmaking system, as well as the lack of systematic assessments of quality decision making and the relatively infrequent use of formal decision-making frameworks. Nevertheless, the results indicate differences as companies and agencies use diverse processes to arrive at the final decision either through consensus, majority vote, or an individual making the decision. The majority of companies and agencies believe that the quality of decision making can and should be measured. Moreover, organizations considered the occurrence of biases within their organization as pertinent. Finally, almost all the participants felt that there was room for improvement for their organization's quality of decision making. These findings are consistent with a published study on regulatory processes and support the need for more consistent and predictable decision-making processes during the life cycle of medicines. This could be achieved through capacity building, systematically evaluating the quality of decision making, and encouraging utilization of formal decision-making frameworks within companies and agencies.


Connecting slow earthquakes to huge earthquakes.
Slow earthquakes are characterized by a wide spectrum of fault slip behaviors and seismic radiation patterns that differ from those of traditional earthquakes. However, slow earthquakes and huge megathrust earthquakes can have common slip mechanisms and are located in neighboring regions of the seismogenic zone. The frequent occurrence of slow earthquakes may help to reveal the physics underlying megathrust events as useful analogs. Slow earthquakes may function as stress meters because of their high sensitivity to stress changes in the seismogenic zone. Episodic stress transfer to megathrust source faults leads to an increased probability of triggering huge earthquakes if the adjacent locked region is critically loaded. Careful and precise monitoring of slow earthquakes may provide new information on the likelihood of impending huge earthquakes.


A novel family of human leukocyte antigen class II receptors may have its origin in archaic human species.
HLA class II α and β chains form receptors for antigen presentation to CD4(+) T cells. Numerous pairings of class II α and β subunits from the wide range of haplotypes and isotypes may form, but most of these combinations, in particular those produced by isotype mixing, yielded mismatched dimers. It is unclear how selection of functional receptors is achieved. At the atomic level, it is not known which interactions of class II residues regulate selection of matched αβ heterodimers and the evolutionary origin of matched isotype mixed dimer formation. In this study we investigated assembly of isotype-mixed HLA class II α and β heterodimers. Assembly and carbohydrate maturation of various HLA-class II isotype-mixed α and β subunits was dependent on the groove binding section of the invariant chain (Ii). By mutation of polymorphic DPβ sequences, we identified two motifs, Lys-69 and GGPM-(84-87), that are engaged in Ii-dependent assembly of DPβ with DRα. We identified five members of a family of DPβ chains containing Lys-69 and GGPM 84-87, which assemble with DRα. The Lys/GGPM motif is present in the DPβ sequence of the Neanderthal genome, and this ancient sequence is related to the human allele DPB1*0401. By site-directed mutagenesis, we inspected Neanderthal amino acid residues that differ from the DPB1*0401 allele and aimed to determine whether matched heterodimers are formed by assembly of DPβ mutants with DRα. Because the *0401 allele is rare in the sub-Saharan population but frequent in the European population, it may have arisen in modern humans by admixture with Neanderthals in Europe.


Unique hybrids between the fungal pathogens Cryptococcus neoformans and Cryptococcus gattii.
Cryptococcus neoformans and Cryptococcus gattii are yeasts that cause meningoencephalitis, but that differ in host range and geographical distribution. Cryptococcus neoformans occurs world-wide and mostly infects immunocompromised patients, whereas C. gattii occurs mainly in (sub)tropical regions and infects healthy individuals. Anomalous C. neoformans strains were isolated from patients. These strains were found to be monokaryotic, and diploid or aneuploid. Amplified Fragment Length Polymorphism (AFLP) and sequence analyses indicated that AFLP genotypes 2 (C. neoformans) and 4 (C. gattii) were present. The strains were serologically BD. Mating- and serotype-specific PCR reactions showed that the strains were MATa-serotype D/MATalpha-serotype B. This study is the first to describe naturally occurring hybrids between C. neoformans and C. gattii.


Functional Connectivity between the Resting-State Olfactory Network and the Hippocampus in Alzheimer's Disease.
Olfactory impairment is associated with prodromal Alzheimer's disease (AD) and is a risk factor for the development of dementia. AD pathology is known to disrupt brain regions instrumental in olfactory information processing, such as the primary olfactory cortex (POC), the hippocampus, and other temporal lobe structures. This selective vulnerability suggests that the functional connectivity (FC) between the olfactory network (ON), consisting of the POC, insula and orbital frontal cortex (OFC) (Tobia et al., 2016), and the hippocampus may be impaired in early stage AD. Yet, the development trajectory of this potential FC impairment remains unclear. Here, we used resting-state functional magnetic resonance imaging (rs-fMRI) data from the Alzheimer's Disease Neuroimaging Initiative (ADNI) to investigate FC changes between the ON and hippocampus in four groups: aged-matched cognitively normal (CN), early mild cognitive impairment (EMCI), late mild cognitive impairment (LMCI), and AD. FC was calculated using low frequency fMRI signal fluctuations in the ON and hippocampus (Tobia et al., 2016). We found that the FC between the ON and the right hippocampus became progressively disrupted across disease states, with significant differences between EMCI and LMCI groups. Additionally, there were no significant differences in gray matter hippocampal volumes between EMCI and LMCI groups. Lastly, the FC between the ON and hippocampus was significantly correlated with neuropsychological test scores, suggesting that it is related to cognition in a meaningful way. These findings provide the first in vivo evidence for the involvement of FC between the ON and hippocampus in AD pathology. Results suggest that functional connectivity (FC) between the olfactory network (ON) and hippocampus may be a sensitive marker for Alzheimer's disease (AD) progression, preceding gray matter volume loss.


Comparative assessments of objective peak-detection algorithms. II. Studies in men.
The performances of eight currently available computerized pulse-detection algorithms were compared on signal-free noise and physiological luteinizing hormone (LH) time series. Signal-free noise was made to vary from 4 to 36% for Gaussian and empirical distributions. Physiological LH data were obtained by immunoassay of blood samples withdrawn every 5 min for 24 h in 8 healthy men, so that the data sets could be emended to simulate varying sampling intensities. Whenever possible, programs were tested at a presumptive 1% false-positive rate. In relation to signal-free noise, the Santen and Bardin program and its modification manifested elevated false-positive rates when the intraseries coefficients of variation increased. The Regional Dual-Threshold program yielded a 1% false-positive rate except on simulated series with high variance. The Cluster and Detect programs both approximated a 1% false-positive rate and the Ultra program approximated a 2.3% false-positive rate throughout the entire range of variance tested. In regard to physiological LH data, all algorithms disclosed a significant impact of sampling intensity on estimates of LH pulse frequency. Sampling-intensity dependent estimates of LH peak frequency by three of the eight programs (Ultra, Cluster, and Detect) were statistically indistinguishable from each other but distinct from the five other programs tested. Furthermore, when judged in relation to their ability to identify individual peaks, the three congruent programs were minimally distinguishable (McNemar's test). Rather, these programs identified the same particular peaks (as defined by concordance of peak maxima) at least 72% of the time.


Magnetic resonance imaging for diagnosing lumbar spinal pathology in adult patients with low back pain or sciatica: a diagnostic systematic review.
In about 5% of all cases LBP is associated with serious underlying pathology requiring diagnostic confirmation and directed treatment. Magnetic resonance imaging (MRI) is often used for this diagnostic purpose yet its role remains controversial. Consequently, this review aimed to summarize the available evidence on the diagnostic accuracy of MRI for identifying lumbar spinal pathology in adult low back pain (LPB) or sciatica patients. MEDLINE, EMBASE and CINAHL were searched (until December 2009) for observational studies assessing the diagnostic accuracy of MRI compared to a reference test for the identification of lumbar spinal pathology. Two reviewers independently selected studies for inclusion, extracted data and assessed methodological quality. Pooled summary estimates of sensitivity and specificity with 95% confidence intervals were calculated for homogenous subsets of studies. Eight studies were included in this review. Strata were defined for separate pathologies i.e. lumbar disc herniation (HNP) and spinal stenosis. Five studies comparing MRI to findings at the surgery for identifying HNP were included in a meta-analysis. Pooled analysis resulted in a summary estimate of sensitivity of 75% (95% CI 65-83%) and specificity of 77% (95% CI 61-88%). For spinal stenosis pooling was not possible. The results suggest that a considerable proportion of patients may be classified incorrectly by MRI for HNP and spinal stenosis. However, the evidence for the diagnostic accuracy of MRI found by this review is not conclusive, since the results could be distorted due to the limited number of studies and large heterogeneity.


The effect of supplementation with fish oil during pregnancy on breast milk immunoglobulin A, soluble CD14, cytokine levels and fatty acid composition.
Breast milk contains many immunomodulatory factors (soluble CD14 (sCD14), IgA and cytokines) with the potential to influence infant immune development. To determine if changes in breast milk omega-3 polyunsaturated fatty acid (n-3 PUFA) composition as a result of maternal dietary fish oil supplementation during pregnancy can modify levels of these immunological parameters in breast milk. In a randomized controlled trial, 83 atopic women received either 4 g fish oil capsules (containing 3.7 g n-3 PUFA) (n = 40) or 4 g olive oil capsules (n = 43) from 20 weeks gestation until delivery. Breast milk was collected 3 days post-partum and fatty acids were analysed by gas liquid chromatography and IgA, sCD14 and cytokines (IL-5, IL-6, IL-10, TNF-alpha and IFN-gamma) were quantitated by ELISA or time resolved fluorescence (TRF). Omega-3 docosahexaenoic acid (DHA; 22:6n-3) and eicosapentaenoic acid (EPA; 20:5n-3) levels were significantly higher (P < 0.001) in breast milk from women supplemented with fish oil (n = 33, DHA mean 1.15%, SD 0.47% and EPA mean 0.16%, SD 0.07%) than in samples from the control group (n = 40, DHA mean 0.50%, SD 0.17% and EPA mean 0.05%, SD 0.02%). Breast milk arachidonic acid (AA; 20:4n-6) levels were significantly lower (P = 0.045) in the fish oil group (mean 0.55%, SD 0.12%) compared with the control group (mean 0.61%, SD 0.14%). Breast milk IgA was positively correlated with DHA (P = 0.046) and 22:5n-3 (P = 0.003), but inversely correlated with linoleic acid (LA; 18:2n-6) (P=0.034). Levels of sCD14 were also positively correlated with 22:5n-3 (P=0.009). Cytokines involved in IgA synthesis (IL-10 and IL-6) were also significantly correlated with both IgA and n-3 PUFA levels, although there were no differences in the levels of breast milk IgA, sCD14 or cytokines between study groups. Supplementation with fish oil during pregnancy significantly alters early post-partum breast milk fatty acid composition. omega-3 PUFA levels were positively associated with IgA and sCD14 levels, suggesting a relationship between fatty acid status and mucosal immune function.


Effects of enriched physical and social environments on motor performance, associative learning, and hippocampal neurogenesis in mice.
We have studied the motor abilities and associative learning capabilities of adult mice placed in different enriched environments. Three-month-old animals were maintained for a month alone (AL), alone in a physically enriched environment (PHY), and, finally, in groups in the absence (SO) or presence (SOPHY) of an enriched environment. The animals' capabilities were subsequently checked in the rotarod test, and for classical and instrumental learning. The PHY and SOPHY groups presented better performances in the rotarod test and in the acquisition of the instrumental learning task. In contrast, no significant differences between groups were observed for classical eyeblink conditioning. The four groups presented similar increases in the strength of field EPSPs (fEPSPs) evoked at the hippocampal CA3-CA1 synapse across classical conditioning sessions, with no significant differences between groups. These trained animals were pulse-injected with bromodeoxyuridine (BrdU) to determine hippocampal neurogenesis. No significant differences were found in the number of NeuN/BrdU double-labeled neurons. We repeated the same BrdU study in one-month-old mice raised for an additional month in the above-mentioned four different environments. These animals were not submitted to rotarod or conditioned tests. Non-trained PHY and SOPHY groups presented more neurogenesis than the other two groups. Thus, neurogenesis seems to be related to physical enrichment at early ages, but not to learning acquisition in adult mice.


Selective serotonin reuptake inhibitor exposure alters osteoblast gene expression and craniofacial development in mice.
Selective serotonin reuptake inhibitor (SSRI) use in pregnancy has been linked to craniofacial birth defects. Little is known about the effects of serotonin or SSRIs on craniofacial development. Here, we provide evidence that citalopram (SSRI) alters the osteogenic profile of murine calvarial cells and leads to craniofacial dysmorphology. We used mouse calvarial pre-osteoblast cells (MC3T3-E1) to study the biochemical profile (microarray and quantitative reverse transcription polymerase chain reactions) after treatment with a titrated dose of citalopram. We used C57BL-6 wild-type breeders to produce litters treated with a clinical dose of citalopram during the third trimester of pregnancy. We used micro-computed tomography and morphometric measures to determine effects on craniofacial development. Controls included untreated cells and age matched untreated litters. We observed decreases in proliferation and increases in alkaline phosphatase activity after citalopram exposure. We confirmed altered expression of genes linked to osteogenesis including Ocn and significant increase in expression of Alp after 7 days of treatment. Our data suggest altered expression of several genes related to craniofacial development (Fgf2, Fgfr2, Tgfβr2 Irs1, Igf1) and statistically significant changes in expression for (Col2a1, Gdf6, Hmox1, and Notch1). We also observed changes in regulation of the serotonin pathway (Sert, Tph1, Tph2, Htr2a, Lrp5) after treatment with citalopram. After in utero exposure to citalopram, mice displayed shorter narrow snouts, more globular skulls and several craniofacial anomalies. Our results provide confirmatory evidence that citalopram exposure is associated with cellular and morphological alterations of the craniofacial complex, which may have important implications for use during pregnancy.


Double-blind, placebo-controlled study of modafinil for fatigue and cognition in schizophrenia patients treated with psychotropic medications.
To assess the effects of modafinil on fatigue, symptoms, attention, working memory, and executive functioning in schizophrenia patients treated with psychotropic medications. Twenty-four patients with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder (10 men and 14 women) were randomly assigned to modafinil up to 200 mg a day (N = 13) or placebo (N = 11) as an adjunct therapy in an 8-week, double-blind, placebo-controlled study. Data were collected from May 18, 2001 to September 11, 2003. Four subjects terminated the study early, including one because of worsening of psychosis during the first week taking modafinil. In the modafinil (N = 10) and placebo (N = 10) groups, fatigue improved significantly over time (p < .01), but there were no differences between groups on changes in fatigue, positive and negative symptoms, or cognition. Fatigue improved in both groups, and there were no differences between groups on changes in fatigue, symptoms, attention, working memory, or executive functioning. Lack of differences between groups may be due to small sample size or possible regression to the mean in the placebo group.


circ_0067934 increases bladder cancer cell proliferation, migration and invasion through suppressing miR-1304 expression and increasing Myc expression levels.
circRNAs have been demonstrated to be key regulators of bladder cancer progression. The present study aimed to investigate the effects of circular RNA (circ)_0067934 in bladder cancer progression. A total of 54 patients with primary bladder cancer were enrolled, and their tumor tissues and adjacent normal bladder tissues were collected. For in vitro functional assays, T24 cells were transfected with sicirc_0067934, and Cell Counting Kit-8 was used to analyze the proliferative capacity of T24 cells. In addition, Transwell and Matrigel assays were used to assess the cell migration and invasion abilities, and a dual-luciferase reporter assay was used to investigate the relationship between miR-1304 and circ_0067934. Finally, reverse transcription-quantitative PCR and western blotting were performed to analyze gene and protein expression levels, respectively. circ_0067934 expression levels were significantly increased in bladder cancer tissues (P<0.001), which was associated with metastasis and a significantly decreased 5-year overall (P<0.05) and disease-free survival (P<0.05). In vitro, T24 cells in the small interfering RNA (si)circ_0067934 group demonstrated significantly reduced proliferation, migration and invasion abilities compared with the si negative control (siNC) group (P<0.01). In addition, the knockdown of circ_0067934 directly increased microRNA (miR)-1304 expression levels in T24 cells. Myc was subsequently discovered to be directly inhibited by miR-1304 and circ_0067934 was observed to increase Myc expression levels in T24 cells through inhibiting miR-1304 expression levels (P<0.01). Compared with the siNC group and sicirc_0067934 + Myc overexpression group, T24 cells in the sicirc_0067934 group exhibited significantly decreased proliferative, migratory and invasive abilities (P<0.01). In conclusion, circ_0067934 was demonstrated to increase bladder cancer cell proliferation, migration and invasion through promoting Myc expression levels via the suppression of miR-1304 expression.


Fractal generalized zone plates.
The construction of fractal generalized zone plates from a set of periodic diffractive optical elements with circular symmetry is proposed. This allows us, for instance, to increase the number of foci of a conventional fractal zone plate while keeping the self-similarity property within the axial irradiance. The focusing properties of these fractal diffractive optical elements for points not only along but also in the close vicinity of the optical axis are investigated. In both cases analytical expressions for the irradiance are derived. Numerical simulations of the energetic efficiency of fractal generalized zone plates under plane wave illumination are carried out. In addition, some effects on the axial irradiance caused by variations in the area of their transparent rings are shown.


Losing ground: physician income, 1995-2003.
Between 1995 and 2003, average physician net income from the practice of medicine declined about 7 percent after adjusting for inflation, according to a national study from the Center for Studying Health System Change (HSC). The decline in physicians' real income stands in sharp contrast to the wage trends for other professionals who saw about a 7 percent increase after adjusting for inflation during the same period. Among different types of physicians, primary care physicians fared the worst with a 10.2 percent decline in real income between 1995 and 2003, while surgeons' real income declined by 8.2 percent. But medical specialists' real income essentially remained unchanged. Physicians reported working slightly fewer hours overall but spent more time on direct patient care. Flat or declining fees from both public and private payers appear to be a major factor underlying declining real incomes for physicians. The downward trend in real incomes since the mid-1990s likely is an important reason for growing physician unwillingness to undertake pro bono work, including charity care and volunteering to serve on hospital committees.


Cluster homes: a community for profoundly and severely retarded persons.
This article discusses community residences that in 1983 replaced two large institutional facilities for retarded persons in Florida. Twenty-six of these residences, or clusters, usually housing 24 clients each, are located throughout the state. The clusters are designed to normalize the living conditions of the mentally retarded, increase their safety, and place them close to society and their families. Medical and social services, including occupational therapy, are made available to all residents of the clusters. This article focuses on one of these clusters, with special emphasis on the role and scope of occupational therapy services within this unique setting. Special problems and experiences encountered in the delivery of occupational therapy services to the profoundly and severely retarded clients in the clusters are explored.


Voluntary collective isolation as a best response to COVID-19 for indigenous populations? A case study and protocol from the Bolivian Amazon.
Indigenous communities worldwide share common features that make them especially vulnerable to the complications of and mortality from COVID-19. They also possess resilient attributes that can be leveraged to promote prevention efforts. How can indigenous communities best mitigate potential devastating effects of COVID-19? In Bolivia, where nearly half of all citizens claim indigenous origins, no specific guidelines have been outlined for indigenous communities inhabiting native communal territories. In this Public Health article, we describe collaborative efforts, as anthropologists, physicians, tribal leaders, and local officials, to develop and implement a multiphase COVID-19 prevention and containment plan focused on voluntary collective isolation and contact-tracing among Tsimane forager-horticulturalists in the Bolivian Amazon. Phase 1 involves education, outreach, and preparation, and phase 2 focuses on containment, patient management, and quarantine. Features of this plan might be exported and adapted to local circumstances elsewhere to prevent widespread mortality in indigenous communities.


Endovascular therapy for critical limb ischemia.
Critical limb ischemia (CLI) represents an advanced disease state of peripheral arterial disease. It manifests as lower extremity ischemic rest pain or ischemic skin lesions leading to ulceration or gangrene. Patients with CLI often have multiple medical comorbidities and a 1-year mortality rate of 25% and a 1-year amputation rate of 25%. Historically, bypass surgery with autogenous veins for flow restoration has been the first-line therapy for CLI. However, advances in endovascular techniques and device technology have changed the treatment paradigm. Catheter-based technologies are rapidly evolving at a rate that is outpacing large-scale studies evaluating relevant clinical outcomes. Patients with CLI require a multidisciplinary management approach centered on aggressive medical therapies, wound care and prompt revascularization, with an emphasis on limb salvage. This review summarizes the contemporary endovascular therapies including balloon angioplasty, atherectomy and bare-metal stenting. In addition, we review emerging technologies, such as drug-eluting stents, drug-coated balloons and chronic total occlusion recanalization devices.


Compensatory water effects link yearly global land CO2 sink changes to temperature.
Large interannual variations in the measured growth rate of atmospheric carbon dioxide (CO2) originate primarily from fluctuations in carbon uptake by land ecosystems. It remains uncertain, however, to what extent temperature and water availability control the carbon balance of land ecosystems across spatial and temporal scales. Here we use empirical models based on eddy covariance data and process-based models to investigate the effect of changes in temperature and water availability on gross primary productivity (GPP), terrestrial ecosystem respiration (TER) and net ecosystem exchange (NEE) at local and global scales. We find that water availability is the dominant driver of the local interannual variability in GPP and TER. To a lesser extent this is true also for NEE at the local scale, but when integrated globally, temporal NEE variability is mostly driven by temperature fluctuations. We suggest that this apparent paradox can be explained by two compensatory water effects. Temporal water-driven GPP and TER variations compensate locally, dampening water-driven NEE variability. Spatial water availability anomalies also compensate, leaving a dominant temperature signal in the year-to-year fluctuations of the land carbon sink. These findings help to reconcile seemingly contradictory reports regarding the importance of temperature and water in controlling the interannual variability of the terrestrial carbon balance. Our study indicates that spatial climate covariation drives the global carbon cycle response.


Mobility Patterns During COVID-19 Travel Restrictions in Nairobi Urban Informal Settlements: Who Is Leaving Home and Why.
Nairobi's urban slums are ill equipped to prevent spread of the novel coronavirus disease (COVID-19) due to high population density, multigenerational families in poorly ventilated informal housing, and poor sanitation. Physical distancing policies, curfews, and a citywide lockdown were implemented in March and April 2020 resulting in sharp decreases in movement across the city. However, most people cannot afford to stay home completely (e.g., leaving daily to fetch water). If still employed, they may need to travel longer distances for work, potentially exposing them COVID-19 or contributing to its spread. We conducted a household survey across five urban slums to describe factors associated with mobility in the previous 24 h. A total of 1695 adults were interviewed, 63% female. Of these, most reported neighborhood mobility within their informal settlement (54%), 19% stayed home completely, and 27% reported long-distance mobility outside their informal settlement, mainly for work. In adjusted multinomial regression models, women were 58% more likely than men to stay home (relative risk ratio (RRR): 1.58, 95% confidence interval (CI): 1.16, 2.14) and women were 60% less likely than men to report citywide mobility (RRR: 0.40; 95% CI 0.31, 0.52). Individuals in the wealthiest quintile, particularly younger women, were most likely to not leave home at all. Those who reported citywide travel were less likely to have lost employment (RRR: 0.49; 95% CI 0.38, 0.65) and were less likely to avoid public transportation (RRR: 0.30; 95% CI 0.23, 0.39). Employment and job hunting were the main reasons for traveling outside of the slum; less than 20% report other reasons. Our findings suggest that slum residents who retain their employment are traveling larger distances across Nairobi, using public transportation, and are more likely to be male; this travel may put them at higher risk of COVID-19 infection but is necessary to maintain income. Steps to protect workers from COVID-19 both in the workplace and while in transit (including masks, hand sanitizer stations, and reduced capacity on public transportation) are critical as economic insecurity in the city increases due to COVID-19 mitigation measures. Workers must be able to commute and maintain employment to not be driven further into poverty. Additionally, to protect the majority of individuals who are only travelling locally within their settlement, mitigation measures such as making masks and handwashing stations accessible within informal settlements must also be implemented, with special attention to the burden placed on women.


Gastric emptying and orocecal transit time in pregnancy.
To evaluate the effects of pregnancy on gastrointestinal function, we determined gastric emptying time, orocecal transit time, and fasting gastrointestinal hormone levels (cholecystokinin, gastrin, pancreatic polypeptide, neurotensin) in 11 women with mild dyspeptic symptoms during the first and third trimesters of their pregnancies, and again 4-6 months after delivery. After the women ingested a disaccharide solution, orocecal transit time was determined by monitoring breath hydrogen concentrations at 10-min intervals, and values were compared with the postpartum value. Ultrasound examinations of gastric emptying were performed during the same intervals. The half-emptying time and the final gastric emptying time did not differ in the first and third trimesters and postpartum, but gastrointestinal transit time was significantly longer in the third trimester of pregnancy than postpartum (100.0 min [range, 50.5-240.0 min] vs 70.0 min [range, 40.5-240.0 min; P < 0.05]), respectively. Mean plasma pancreatic polypeptide values were lower in the third trimester of pregnancy than postpartum. and a negative correlation was observed between pancreatic polypeptide levels and transit time in the third trimester (r = -0.65; P = 0.0261). The plasma levels of other gastrointestinal hormones did not differ in the various periods studied. Our study shows that, despite evident dyspeptic symptoms, there were no significant alterations in gastric emptying or orocecal transit time during the first trimester of pregnancy. Conversely, in the third trimester, orocecal transit time was significantly longer.


Identification of neurotoxic cytokines by profiling Alzheimer's disease tissues and neuron culture viability screening.
Alzheimer's disease (AD) therapeutics based on the amyloid hypothesis have shown minimal efficacy in patients, suggesting that the activity of amyloid beta (Aβ) represents only one aspect of AD pathogenesis. Since neuroinflammation is thought to play an important role in AD, we hypothesized that cytokines may play a direct role in promoting neuronal death. Here, we profiled cytokine expression in a small cohort of human AD and control brain tissues. We identified AD-associated cytokines using partial least squares regression to correlate cytokine expression with quantified pathologic disease state and then used neuron cultures to test whether cytokines up-regulated in AD tissues could affect neuronal viability. This analysis identified cytokines that were associated with the pathological severity. Of the top correlates, only TNF-α reduced viability in neuron culture when applied alone. VEGF also reduced viability when applied together with Aβ, which was surprising because VEGF has been viewed as a neuro-protective protein. We found that this synthetic pro-death effect of VEGF in the context of Aβ was commensurate with VEGFR-dependent changes in multiple signaling pathways that govern cell fate. Our findings suggest that profiling of tissues combined with a culture-based screening approach can successfully identify new mechanisms driving neuronal death.


Molecular characterization of the Trichomonas gallinae morphologic complex in the United States.
Forty-two Trichomonas gallinae isolates were molecularly characterized to determine whether isolates differed in genetic sequence of multiple gene targets depending on host species or geographical location. The 5.8S ribosomal RNA (rRNA) and flanking internal transcribed spacer (ITS) gene regions were amplified by polymerase chain reaction, and the sequences were analyzed phylogenetically. The results of the sequence analysis strongly suggest at least 2 species may exist within the T. gallinae morphologic complex. Based on ITS sequences, one group demonstrated high nucleotide identity to the 3 T. gallinae sequences available in GenBank, whereas the second group was more closely related to T. vaginalis (98%) than to T. gallinae (92%). Two common ground-dove (Columbina passerina) isolates shared a 95% identity with T. vaginalis and a 92% identity with T. gallinae and T. tenax. Sequence analysis of both the 18S rRNA and alpha-tubulin genes from a subset of the isolates supports the 5.8S-ITS sequence results. All of the T. vaginalis-like isolates originated from Arizona, California, or Texas, whereas T. gallinae isolates were found in all sampled states. Both T. vaginalis-like and T. gallinae isolates were involved in trichomoniasis outbreaks in California and Arizona.


Steroidomics for highlighting novel serum biomarkers of testosterone doping.
Aim: Quantification of testosterone (T) and 5α-dihydrotestosterone serum concentrations proved to be an efficient alternative to urinary steroid profiling for the detection of T doping. In this context, additional serum markers could be discovered by exploratory untargeted steroidomics studies. Results: Endogenous steroid metabolites were monitored by ultra high-performance liquid chromatography coupled to high-resolution mass spectrometry in serum samples collected during a T administration clinical trial. A three-step workflow for accurate review of annotation was used and multifactorial data analysis allowed highlighting promising serum biomarkers. Longitudinal monitoring of selected compounds was performed to assess T abuse detection capabilities. Conclusion: Application of serum steroidomics showed high potential for biomarker discovery of T doping, suggesting longitudinal monitoring of steroid hormones in serum as a significant improvement in detection of endogenous steroids abuse.


NetCTLpan: pan-specific MHC class I pathway epitope predictions.
Reliable predictions of immunogenic peptides are essential in rational vaccine design and can minimize the experimental effort needed to identify epitopes. In this work, we describe a pan-specific major histocompatibility complex (MHC) class I epitope predictor, NetCTLpan. The method integrates predictions of proteasomal cleavage, transporter associated with antigen processing (TAP) transport efficiency, and MHC class I binding affinity into a MHC class I pathway likelihood score and is an improved and extended version of NetCTL. The NetCTLpan method performs predictions for all MHC class I molecules with known protein sequence and allows predictions for 8-, 9-, 10-, and 11-mer peptides. In order to meet the need for a low false positive rate, the method is optimized to achieve high specificity. The method was trained and validated on large datasets of experimentally identified MHC class I ligands and cytotoxic T lymphocyte (CTL) epitopes. It has been reported that MHC molecules are differentially dependent on TAP transport and proteasomal cleavage. Here, we did not find any consistent signs of such MHC dependencies, and the NetCTLpan method is implemented with fixed weights for proteasomal cleavage and TAP transport for all MHC molecules. The predictive performance of the NetCTLpan method was shown to outperform other state-of-the-art CTL epitope prediction methods. Our results further confirm the importance of using full-type human leukocyte antigen restriction information when identifying MHC class I epitopes. Using the NetCTLpan method, the experimental effort to identify 90% of new epitopes can be reduced by 15% and 40%, respectively, when compared to the NetMHCpan and NetCTL methods. The method and benchmark datasets are available at http://www.cbs.dtu.dk/services/NetCTLpan/.


Development and validation of a LC-MS/MS method for the simultaneous determination of aflatoxins, dyes and pesticides in spices.
Based on several alerts from European countries over the last years concerning spices, we have been encouraged to establish an accurate method for the determination of dyes, aflatoxins and pesticides in various types of spices using reversed-phase (RP) liquid chromatography-tandem mass spectrometry interfaced with electrospray (LC-ESI-MS/MS). A simple sample treatment procedure entailing the use of an extraction step with acetonitrile without further cleanup has been developed. A C18 column with an aqueous ammonium formate/methanol mixture as the mobile phase was used, and gradient elution was performed. Mass spectral acquisition was done in positive ion mode by applying multiple reaction monitoring of at least two fragmentation transitions per compound to provide a high degree of selectivity. The method was in-house validated in terms of linearity, sensitivity, repeatability, recovery and selectivity on six kinds of spices. Satisfactory results in the majority of the cases were obtained for all analytes and matrices, with practical limits of quantitation acceptable for routine monitoring purposes. Extraction recoveries for most of the compounds ranged from 60% to 140% at spiking levels of 0.05 and 0.5 mg kg(-1). The applicability of the method for the simultaneous determination of dyes, aflatoxins and pesticides in several types of spices was demonstrated, and the method successfully applied to a limited number of products from the local market.


Neural evidence for distracter suppression during visual search in real-world scenes.
Selecting visual information from cluttered real-world scenes involves the matching of visual input to the observer's attentional set--an internal representation of objects that are relevant for current behavioral goals. When goals change, a new attentional set needs to be instantiated, requiring the suppression of the previous set to prevent distraction by objects that are no longer relevant. In the present fMRI study, we investigated how such suppression is implemented at the neural level. We measured human brain activity in response to natural scene photographs that could contain objects from (1) a currently relevant (target) category, (2) a previously but not presently relevant (distracter) category, and/or (3) a never relevant (neutral) category. Across conditions, multivoxel response patterns in object-selective cortex carried information about objects present in the scenes. However, this information strongly depended on the task relevance of the objects. As expected, information about the target category was significantly increased relative to the neutral category, indicating top-down enhancement of task-relevant information. Importantly, information about the distracter category was significantly reduced relative to the neutral category, indicating that the processing of previously relevant objects was suppressed. Such active suppression at the level of high-order visual cortex may serve to prevent the erroneous selection of, or interference from, objects that are no longer relevant to ongoing behavior. We conclude that the enhancement of relevant information and the suppression of distracting information both contribute to the efficient selection of visual information from cluttered real-world scenes.


The Activity of CCL18 is Principally Mediated through Interaction with Glycosaminoglycans.
The CC chemokine ligand 18 (CCL18) was first identified as a chemoattractant for naïve T cells. It has been reported to recruit T and B lymphocytes, and we show here, natural killer (NK) cells, but with low efficacy. Investigation of its ability to elicit G-protein-coupled signaling showed that it does not involve extracellular signal-regulated kinase (ERK) phosphorylation, and it is not able to induce receptor internalization, as assessed on CCR3. CCL18 has recently been reported to possess activities unrelated to cellular recruitment, but it had no effect on T lymphocyte proliferation. We postulated that a more potent chemoattractant may be produced under inflammatory conditions but only minor truncations were observed, with the major form being the full-length protein. In view of the lack of potent immunomodulatory properties, we wondered if binding to CCL18 by the tick chemokine binding proteins Evasin-1 and -4 was an artifact of the methods used, but complex formation was confirmed by size exclusion chromatography, and abrogation of its binding to, and antagonism of, CCR3. Its receptor has remained elusive since its cloning in 1997, although it has been reported to induce migration of breast cancer cells by signaling through PITPNM3, but we show that this receptor is not expressed on lymphocytes. We have developed a radiolabeled equilibrium competition binding assay and demonstrated that it bound with high affinity to peripheral blood leukocytes (PBLs), but the binding was displaced similarly by both unlabelled CCL18 as well as heparin. Both heparin binding and binding to PBLs are considerably abrogated by mutation of the BBXB motif in the 40s loop suggesting an essential role of the CCL18-glycosaminoglycan interaction.


Surface enhanced hyper Raman scattering (SEHRS) and its applications.
Surface enhanced hyper Raman scattering (SEHRS) is the spontaneous, two-photon excited Raman scattering that occurs for molecules residing in high local optical fields of plasmonic nanostructures. Being regarded as a non-linear analogue of surface enhanced Raman scattering (SERS), SEHRS shares most of its properties, but also has additional characteristics. They include complementary spectroscopic information resulting from different selection rules and a stronger enhancement due to the non-linearity in excitation. In practical spectroscopy, this can translate to advantages, which include a high selectivity when probing molecule-surface interactions, the possibility of probing molecules at low concentrations due to the strong enhancement, and the advantages that come with excitation in the near-infrared. In this review, we give examples of the wealth of vibrational spectroscopic information that can be obtained by SEHRS and discuss work that has contributed to understanding the effect and that therefore provides directions for SEHRS spectroscopy. Future applications could range from biophotonics to materials research.


The Association of Cesarean Skin Incision Length and Postoperative Wound Complications.
This study was aimed to evaluate the relationship between cesarean skin incision length and wound complications. Planned secondary analysis of a multicenter double-blind randomized trial of adjunctive azithromycin versus placebo (in addition to standard cefazolin) in women ≥24 weeks undergoing cesarean delivery during labor or ≥4 hours after membrane rupture. Skin incision length (cm) was measured just prior to skin closure. The primary outcome was a composite of wound complications (wound infection, separation, seroma, hematoma, or dehiscence) up to 6 weeks of postpartum. Individual components of the composite were examined as secondary outcomes. Outcomes were compared between groups defined by the lowest (≤25th), middle (25-75th) and highest (>75th) incision length quartiles. Logistic regression was used to adjust for potential confounding variables. Of the 2,013 women enrolled in the primary trial, 1,916 had recorded incision lengths and were included in this secondary analysis. The overall rate of composite wound complications was 7.8%. Median incision length was 15.0 cm (interquartile range: 14.0-16.5) with the lowest quartile defined as ≤14, middle as >14 to ≤16.5, and highest as >16.5 cm. Mean BMI, parity, use of staples, and duration of surgery differed significantly between the three incision length groups. In unadjusted analysis, the longest incision lengths were associated with an increased risk of the wound composite and wound infections (odds ratio [OR] = 2.27, 95% confidence interval [CI]: 1.43-3.60 and OR = 2.30, 95% CI: 1.27-4.15, respectively) compared with the shortest incision lengths. However, after multivariable adjustments, these associations were nullified. Additional analyses considering incision length as a continuous variable and using 10th/90th percentile cut-offs still did not suggest any associations with outcomes. Increasing skin incision length is not independently associated with an increased risk of postoperative wound complications. · After multivariable adjustments, skin incision length was not independently associated with an increased risk of postoperative wound complications.. · A reasonable incision length needed to safely perform the procedure should be used..


Improving tolerability of lithium with a once-daily dosing schedule.
Lithium has been proved to be highly efficacious in the treatment of bipolar affective disorder, though a narrow therapeutic index and a high incidence of troublesome side effects often leads to poor compliance in patients. Therefore, there is a need to explore treatment strategies to improve the efficacy and side effect profile of lithium. We compared the efficacy and side effect profile of a once-daily versus twice-daily dosing schedule of lithium in mania. Eighty-three manic patients according to International Classification of Diseases, 10th Revision, Diagnostic Criteria for Research, giving informed consent were randomly allocated to receive regular lithium carbonate once daily or twice daily. They were assessed using Bech-Rafaelsen Mania Rating Scale, a lithium side effect scale, hemogram, renal function test, lipid profile, and a thyroid function test at baseline and Day 7, Day 21, and Day 42. Repeated-measures analysis of variance for Bech-Rafaelsen Mania Rating Scale scores showed a significant main effect, but interaction of treatment groups over time was not significant. Those subjects receiving twice-daily lithium experienced significantly higher urinary frequency on Day 21 (P = 0.008) and Day 42 (P = 0.035). They also required significantly higher total daily dose of lithium (P = 0.017) and had lower serum lithium levels (P < 0.001). There was a significant positive correlation between urinary frequency at Day 42 with lithium dose. A twice-daily dose of lithium was of similar efficacy as the once-daily schedule but produces higher renal adverse effects that may be dose-related. Therefore, a single daily dose of lithium can be a viable method to reduce the side effects of lithium, which may lead to better patient compliance.


Outpatient parenteral antimicrobial therapy-treated bone and joint infections in a tropical setting.
Osteoarticular infections are a primary indication for outpatient parenteral antimicrobial therapy (OPAT). The climate and geographical diversity of tropical Australia, together with the prevalence of melioidosis, disseminated gonococcal disease and community-acquired methicillin-resistant Staphylococcus aureus renders this a challenging environment in which to manage such infections. We evaluated patients managed by the Royal Darwin Hospital Hospital in the Home service for bone and joint infections. A retrospective analysis of the therapeutic outcomes at the end of intravenous therapy was carried out for patients treated between 1 January 2006 and 15 September 2007. Fifty-five patients were treated, including 21 (38%) indigenous Australians and 18 (33%) from remote communities. Baseline characteristics were similar to other published data, but there were two cases each of gonococcal septic arthritis and melioidosis. During treatment, 39 (71%) lived at home, with five (9%) of these receiving treatment at community clinics. Thirteen (24%) resided in self-care units in the hospital grounds. Three (5%) were managed at hostels or in prison. Median duration of parenteral therapy was 42 days, with a median of 22 days outside hospital, providing a total saving of 1307 bed-days. Clinical success at end of therapy was 84%, with no significant difference between indigenous and non-indigenous cohorts. OPAT for osteoarticular infections is both feasible and effective in a tropical environment, including for indigenous patients. Extension of treatment to remote-dwelling patients is facilitated by the innovative use of self-care units and administration of treatment at remote clinics.


Molecular energy dissipation in nanoscale networks of Dentin Matrix Protein 1 is strongly dependent on ion valence.
The fracture resistance of biomineralized tissues such as bone, dentin, and abalone is greatly enhanced through the nanoscale interactions of stiff inorganic mineral components with soft organic adhesive components. A proper understanding of the interactions that occur within the organic component, and between the organic and inorganic components, is therefore critical for a complete understanding of the mechanics of these tissues. In this paper, we use Atomic Force Microscope (AFM) force spectroscopy and dynamic force spectroscopy to explore the effect of ionic interactions within a nanoscale system consisting of networks of Dentin Matrix Protein 1 (DMP1) (a component of both bone and dentin organic matrix), a mica surface, and an AFM tip. We find that DMP1 is capable of dissipating large amounts of energy through an ion-mediated mechanism, and that the effectiveness increases with increasing ion valence.


Restoration of torque in defective flagellar motors.
Paralyzed motors of motA and motB point and deletion mutants of Escherichia coli were repaired by synthesis of wild-type protein. As found earlier with a point mutant of motB, torque was restored in a series of equally spaced steps. The size of the steps was the same for both MotA and MotB. Motors with one torque generator spent more time spinning counterclockwise than did motors with two or more generators. In deletion mutants, stepwise decreases in torque, rare in point mutants, were common. Several cells stopped accelerating after eight steps, suggesting that the maximum complement of torque generators is eight. Each generator appears to contain both MotA and MotB.


The usefulness of radiological grading scales to predict pain intensity, functional impairment, and health-related quality of life after surgery for lumbar degenerative disc disease.
The goal of this study is to determine the relationship of radiological grading scales of lumbar degenerative disc disease (DDD) with postoperative pain intensity, functional impairment, and health-related quality of life (HRQoL). Response to surgical treatment at 6 weeks (W6) on the visual analogue scale (VAS) for back and leg pain, Oswestry-Disability (ODI) and Roland-Morris Disability Index (RMDI), Timed Up and Go (TUG) test, EuroQol (EQ) 5D, and Short-Form Health-Survey (SF-12) physical component summary (PCS) was compared between patients with different Modic (MOD) and Pfirrmann (PFI) grades. Longitudinal outcomes at day 3 (D3), W6, 6 months (M6), and 1 year (Y1) were compared. The study included 338 patients (mean age, 58.6 years), of which n = 202 (59.8%) had MOD 1-3 and n = 217 (64.2%) PFI 4-5 changes. Patients with MOD 1-3 were as likely as patients without MOD changes to be treatment-responders at W6 in terms of VAS leg pain, ODI, RMDI, TUG, EQ5D, and SF-12 PCS. Similarly, patients with PFI 4-5 were as likely as patients with PFI 1-3 changes to be treatment-responders at W6. Longitudinal outcomes were similar at D3, W6, M6, and Y1 between patients with and without MOD changes. Patients with PFI 4-5 fared similar to those with PFI 1-3 except for inferior HRQoL on the SF-12 PCS metric at Y1. There was no distinct relationship between commonly used radiological grading scales of lumbar DDD with clinical outcome. Therefore, no prognosis should be made on the grounds of preoperative PFI and MOD classifications for patients undergoing spine surgery for lumbar DDD.


Structural basis of substrate specificity and protease inhibition in Norwalk virus.
Norwalk virus (NV), the prototype human calicivirus, is the leading cause of nonbacterial acute gastroenteritis. The NV protease cleaves the polyprotein encoded by open reading frame 1 of the viral genome at five nonhomologous sites, releasing six nonstructural proteins that are essential for viral replication. The structural details of how NV protease recognizes multiple substrates are unclear. In our X-ray structure of an NV protease construct, we observed that the C-terminal tail, representing the native substrate positions P5 to P1, is inserted into the active site cleft of the neighboring protease molecule, providing atomic details of how NV protease recognizes a substrate. The crystallographic structure of NV protease with the C-terminal tail redesigned to mimic P4 to P1 of another substrate site provided further structural details on how the active site accommodates sequence variations in the substrates. Based on these structural analyses, substrate-based aldehyde inhibitors were synthesized and screened for inhibition potency. Crystallographic structures of the protease in complex with each of the three most potent inhibitors were determined. These structures showed concerted conformational changes in the S4 and S2 pockets of the protease to accommodate variations in the P4 and P2 residues of the substrate/inhibitor, which could be a mechanism for how the NV protease recognizes multiple sites in the polyprotein with differential affinities during virus replication. These structures further indicate that the mechanism of inhibition by these inhibitors involves covalent bond formation with the side chain of the conserved cysteine in the active site by nucleophilic addition, and such substrate-based aldehydes could be effective protease inhibitors.


The genome sequence and effector complement of the flax rust pathogen Melampsora lini.
Rust fungi cause serious yield reductions on crops, including wheat, barley, soybean, coffee, and represent real threats to global food security. Of these fungi, the flax rust pathogen Melampsora lini has been developed most extensively over the past 80 years as a model to understand the molecular mechanisms that underpin pathogenesis. During infection, M. lini secretes virulence effectors to promote disease. The number of these effectors, their function and their degree of conservation across rust fungal species is unknown. To assess this, we sequenced and assembled de novo the genome of M. lini isolate CH5 into 21,130 scaffolds spanning 189 Mbp (scaffold N50 of 31 kbp). Global analysis of the DNA sequence revealed that repetitive elements, primarily retrotransposons, make up at least 45% of the genome. Using ab initio predictions, transcriptome data and homology searches, we identified 16,271 putative protein-coding genes. An analysis pipeline was then implemented to predict the effector complement of M. lini and compare it to that of the poplar rust, wheat stem rust and wheat stripe rust pathogens to identify conserved and species-specific effector candidates. Previous knowledge of four cloned M. lini avirulence effector proteins and two basidiomycete effectors was used to optimize parameters of the effector prediction pipeline. Markov clustering based on sequence similarity was performed to group effector candidates from all four rust pathogens. Clusters containing at least one member from M. lini were further analyzed and prioritized based on features including expression in isolated haustoria and infected leaf tissue and conservation across rust species. Herein, we describe 200 of 940 clusters that ranked highest on our priority list, representing 725 flax rust candidate effectors. Our findings on this important model rust species provide insight into how effectors of rust fungi are conserved across species and how they may act to promote infection on their hosts.


A novel approach to identify genes that determine grain protein deviation in cereals.
Grain yield and protein content were determined for six wheat cultivars grown over 3 years at multiple sites and at multiple nitrogen (N) fertilizer inputs. Although grain protein content was negatively correlated with yield, some grain samples had higher protein contents than expected based on their yields, a trait referred to as grain protein deviation (GPD). We used novel statistical approaches to identify gene transcripts significantly related to GPD across environments. The yield and protein content were initially adjusted for nitrogen fertilizer inputs and then adjusted for yield (to remove the negative correlation with protein content), resulting in a parameter termed corrected GPD. Significant genetic variation in corrected GPD was observed for six cultivars grown over a range of environmental conditions (a total of 584 samples). Gene transcript profiles were determined in a subset of 161 samples of developing grain to identify transcripts contributing to GPD. Principal component analysis (PCA), analysis of variance (ANOVA) and means of scores regression (MSR) were used to identify individual principal components (PCs) correlating with GPD alone. Scores of the selected PCs, which were significantly related to GPD and protein content but not to the yield and significantly affected by cultivar, were identified as reflecting a multivariate pattern of gene expression related to genetic variation in GPD. Transcripts with consistent variation along the selected PCs were identified by an approach hereby called one-block means of scores regression (one-block MSR).


Orthotopic liver transplantation in patients with human immunodeficiency virus and end-stage liver disease.
Patients with human immunodeficiency virus (HIV) most often have hepatitis C virus (HCV) or hepatitis B (HBV) virus coinfection, or both, as a cause of their liver disease. Recent survival statistics show that patients infected with HIV treated with highly active antiretroviral therapy (HAART) can expect a significant prolongation of life by interfering with the natural progression of HIV to acquired immune deficiency syndrome (AIDS). Therefore, HIV-positive patients experiencing complications of liver failure are at greater immediate risk of dying from their end-stage liver disease (ESLD) rather than their HIV. Many transplant centers still consider HIV infection as a contraindication for orthotopic liver transplantation (OLT). At our two institutions, we believe that patients with HIV suffering from ESLD should be considered for OLT. This study evaluates the survival of patients undergoing OLT with HIV under HAART therapy. OLT was performed in 16 patients with HIV suffering from ESLD as a result of chronic HCV, chronic HBV, or fulminant hepatic failure (FHF). Collected data include patient demographics, patient and graft survival, pre-OLT assessments, and postoperative complications (including opportunistic infections). Ten patients at Pittsburgh and 6 patients at Miami received OLT. Of the 16 patients who received OLT, 14 remain alive to date. Thirteen of 16 patients are more than 12 months post-OLT, whereas the last patient is currently 6 months post-OLT. Five patients at Miami and 9 of 10 patients at Pittsburgh received HAART therapy before OLT, although 2 of the Pittsburgh patients had their HAART therapy discontinued before OLT because of significant liver dysfunction. The pre-OLT viral loads were undetectable in 13 of 16 patients. The cluster determinant (CD)4 count was less than 200 in 6 patients and greater than 100 in 2 patients before OLT. In all patients, CD4 counts increased above 200 in the post-OLT period. Tacrolimus toxicity associated with the pharmacologic inhibition of cytochrome p450 metabolism caused by protease inhibitors occurred in 6 patients after OLT. Six patients (38%) experienced acute cellular rejection immediately after OLT. Our experience suggests that OLT is effective in selected HIV-positive patients suffering from ESLD. Patient and graft survival was similar to non-HIV-positive patients suffering from the same indications for OLT. Acute cellular rejection was no less frequent that seen in non-HIV-positive patients. Given the complex pharmacologic interactions between the protease inhibitors and tacrolimus, careful monitoring, and attention is required to prevent toxicity or underdosing.


Surgical debridement and primary closure of the oral mucosa for repair of open mandibular pars incisiva fractures in three neonatal calves.
3 neonatal female calves (ages, < 1 to 4 days) were examined because of mandibular trauma. Physical examination indicated that each calf had an open fracture of the mandibular pars incisiva (rostral mandibular fracture) with ventral displacement of the incisors at the affected region. Oral radiographs were obtained for 1 calf and revealed that 5 incisors were fractured at the level of the apical dental buds. Each calf was anesthetized. The fracture site and surrounding tissues were surgically debrided and flushed with sterile 0.05% chlorhexidine solution. The laceration in the oral mucosa was closed with absorbable suture in an interrupted horizontal mattress pattern. Additionally, a Penrose drain was placed during primary closure and removed 4 days later in 1 calf. The fractured incisors were removed during primary debridement in another calf. All calves received perioperative antimicrobials and analgesics. One calf developed mild osteomyelitis of the rostral mandible, which resolved with additional surgical debridement and antimicrobial treatment. That calf and another developed mild brachygnathia. At the time of last follow-up (3 to 13 months after hospital discharge), all 3 calves were eating and growing as expected. 3 calves with open rostral mandibular fractures were successfully managed by surgical debridement and primary closure of the oral laceration. The procedure was easy to perform, did not require specialized equipment, and was less expensive than other repair methods. This procedure may be an effective and economic on-farm treatment alternative for calves with rostral mandibular fractures.


Breast cancer incidence and mortality in the Nordic capitals, 1970-1998. Trends related to mammography screening programmes.
The aim of the present study was to relate the time trends in breast cancer incidence and mortality to the introduction of mammography screening in the Nordic capitals. Helsinki offered screening to women aged 50-59 starting in 1986. The other three capitals offered screening to women aged 50-69 starting in 1989 in Stockholm, 1991 in Copenhagen, and 1996 in Oslo. Prevalence peaks in breast cancer incidence depended on the age groups covered by the screening, the length of the implementation of screening, and the extent of background opportunistic screening. No mortality reduction following the introduction of screening was visible after seven to 12 years of screening in any of the three capitals where significant effects of the screening on the breast cancer mortality had already been demonstrated by using other analytical methods for the evaluation. No visible effect on mortality reduction was expected in Oslo due to too short an observation period. The study showed that the population-based breast cancer mortality trend is too crude a measure to detect the effect of screening on breast cancer mortality during the first years after the start of a programme.


The etiology of autistic traits in preschoolers: a population-based twin study.
Autism Spectrum Disorders (ASD) are highly heritable, but the exact etiological mechanisms underlying the condition are still unclear. Using a multiple rater twin design in a large sample of general population preschool twins, this study aimed to (a) estimate the contribution of genetic and environmental factors to autistic traits, controlling for the possible effects of rater bias, (b) to explore possible sex differences in etiology and (c) to investigate the discordance in autistic traits in monozygotic and same-sex dizygotic twin pairs. The Netherlands Twin Register collected maternal and paternal ratings on autistic traits from a general population of 38,798 three-year-old twins. Autistic traits were assessed with the DSM-oriented Pervasive Developmental Problems scale of the Child Behavior Check List for preschoolers (1½-5 years). Mother and fathers showed high agreement in their assessment of autistic traits (r = .60-.66). Differences between children in autistic traits were largely accounted for by genetic effects (boys: 78% and girls: 83%). Environmental effects that are unique to a child also played a modest role. Environmental effects shared by children growing up in the same family were negligible, once rater bias was controlled for. While the prevalence for clinical ASD is higher in boys than in girls, this study did not find evidence for striking differences in the etiology of autistic traits across the sexes. Even though the heritability was high, 29% of MZ twin pairs were discordant for high autistic traits (clinical range vs. normal development), suggesting that despite high genetic risk, environmental factors might lead to resilience, unaffected status in the context of genetic risk, in some children. It is important to focus future research on risk factors that might interplay with a genetic disposition for ASD, but also on protective factors that make a difference in the lives of children at genetic risk.


Cytoplasmic tail-dependent internalization of membrane-type 1 matrix metalloproteinase is important for its invasion-promoting activity.
Membrane-type 1 matrix metalloproteinase (MT1-MMP) is an integral membrane proteinase that degrades the pericellular extracellular matrix (ECM) and is expressed in many migratory cells, including invasive cancer cells. MT1-MMP has been shown to localize at the migration edge and to promote cell migration; however, it is not clear how the enzyme is regulated during the migration process. Here, we report that MT1-MMP is internalized from the surface and that this event depends on the sequence of its cytoplasmic tail. Di-leucine (Leu571-572 and Leu578-579) and tyrosine573 residues are important for the internalization, and the mu2 subunit of adaptor protein 2, a component of clathrin-coated pits for membrane protein internalization, was found to bind to the LLY573 sequence. MT1-MMP was internalized predominantly at the adherent edge and was found to colocalize with clathrin-coated vesicles. The mutations that disturb internalization caused accumulation of the enzyme at the adherent edge, though the net proteolytic activity was not affected much. Interestingly, whereas expression of MT1-MMP enhances cell migration and invasion, the internalization-defective mutants failed to promote either activity. These data indicate that dynamic turnover of MT1-MMP at the migration edge by internalization is important for proper enzyme function during cell migration and invasion.


Optimal method for short-term or long-term islet preservation: comparison of islet culture, cold preservation and cryopreservation.
Islet preservation plays an important role for the success of islet transplantation. To determine the optimal method for islet preservation, we compared the outcomes of islet culture, cold preservation, and cryopreservation in this study. Isolated rat islets were divided into three groups: 37 °C group (conventional culture at 37 °C in RPMI-1640 medium), 4 °C group (cold preservation at 4 °C in University of Wisconsin (UW) solution), and -80 °C group (cryopreservation at -80 °C with dimethyl sulfoxide (DMSO)). Recovery rate, Calcein-AM/PI double staining, insulin release, mRNA level of hypoxia-inducible factor-1α (HIF-1α), and protein level of Bax in islets were examined after short-term (1 day) or long-term (7 days) preservation. After either short-term or long-term preservation, 4 °C group showed higher recovery rate of the islets number, lower percentage of PI positive area, better insulin release ability, and lower expression levels of HIF-1α and Bax in comparison to the 37 or -80 °C group. Meanwhile, islets in 37 °C group showed better function, and down-regulation of HIF-1α and Bax than those in -80 °C group on day 1; however, worse function of islets, up-regulated HIF-1α and Bax in 37 °C group were observed in comparison to -80 °C group on day 7. These results suggest that cold preservation at 4 °C in UW solution is the optimal method in comparison to the conventional culture at 37 °C or cryopreservation at -80 °C for short-term or long-term islet preservation. Furthermore, the potential mechanism may relate to, at least in part, apoptosis induced by the HIF-1α.


Evaluation of the intrinsic innervation of the internal anal sphincter using electrical stimulation.
Stimulation of the rectal mucosa with a bipolar electrode leads to relaxation of the internal anal sphincter. Intraoperative studies in two subjects showed that transmission of the impulse was independent of extrinsic nerves and was interrupted by circular myotomy. Characteristics of the reflex were studied in 11 healthy women and 19 women with severe idiopathic constipation. One control subject and two patients did not tolerate the test. In the remainder the stimulus caused a clearly defined fall in internal sphincter pressure. The mean resting maximum anal canal pressure before stimulation was the same in both groups (90 (10) v 104 (7) cm H2O, p = 0.3, controls v patients). The threshold stimulus for relaxation (12 (2) v 14 (1) mamps, p = 0.5), the maximum percent fall in resting pressure (43 (7) v 46 (4)%, p = 0.7) and the lowest absolute resting pressure produced by stimulation (48 (13) v 49 (6) cm H2O, p = 0.9) were the same in both groups. The stimulus required to achieve maximum relaxation was significantly higher in the patient group (23 (3) v 32 (2) mamps, p = 0.012) suggesting abnormal intrinsic innervation of the sphincter in these patients. Electrical stimulation should not replace balloon distension for routine testing of the rectoanal reflex but it may be useful in quantitative studies.


Systemic allergic reactions to gelatin included in vaccines as a stabilizer.
Most of the children who showed systemic immediate-type reactions, including anaphylactic shock, to measles, mumps, rubella, and varicella vaccines had IgE antibodies to gelatin; thus we suspected that the allergic symptoms are caused by gelatin antigen, which is usually included in these live-virus vaccines as a stabilizer. We hypothesized that the anti-gelatin IgE is elicited by immunization with DTaP (diphtheria-tetanus-acellular pertussis) vaccines, which contained a small amount of gelatin as a spillover protein after purification of pertussis toxin. To test this hypothesis, we conducted a case-control study to determine whether children with anti-gelatin IgE had received gelatin-containing DTaP vaccines, and it was indeed found that all such children in the study had immunization histories that included the gelatin-containing DTaP vaccines. Based on these findings, the vaccine manufacturers had removed gelatin from all the DTaP and live-virus vaccines produced in Japan by 2000.


An Analysis of the Changes in Communication Techniques in the Italian Codes of Medical Deontology.
The code of deontology of the Italian National Federation of the Colleges of Physicians, Surgeons and Dentists (FNOMCeO) contains the principles and rules to which the professional medical practitioner must adhere. This work identifies and analyzes the medical-linguistic choices and the expressive techniques present in the different editions of the code, and evaluates their purpose and function, focusing on the first appearance and the subsequent frequency of key terms. Various aspects of the formal and expressive revisions of the eight editions of the Codes of Medical Deontology published after the Second World War (from 1947/48 to 2014) are here presented, starting from a brief comparison with the first edition of 1903. Formal characteristics, choices of medical terminology and the introduction of new concepts and communicative attitudes are here identified and evaluated. This paper, in presenting a quantitative and epistemological analysis of variations, modifications and confirmations in the different editions of the Italian code of medical deontology over the last century, enucleates and demonstrates the dynamic paradigm of changing attitudes in the medical profession. This analysis shows the evolution in medical-scientific communication as embodied in the Italian code of medical deontology. This code, in its adoption, changes and adaptations, as evidenced in its successive editions, bears witness to the expressions and attitudes pertinent to and characteristic of the deontological stance of the medical profession during the twentieth century.


Capsaicin: From Plants to a Cancer-Suppressing Agent.
Capsaicinoids are plant secondary metabolites, capsaicin being the principal responsible for the pungency of chili peppers. It is biosynthesized through two pathways involved in phenylpropanoid and fatty acid metabolism. Plant capsaicin concentration is mainly affected by genetic, environmental and crop management factors. However, its synthesis can be enhanced by the use of elicitors. Capsaicin is employed as food additive and in pharmaceutical applications. Additionally, it has been found that capsaicin can act as a cancer preventive agent and shows wide applications against various types of cancer. This review is an approach in contextualizing the use of controlled stress on the plant to increase the content of capsaicin, highlighting its synthesis and its potential use as anticancer agent.


SBOLCanvas: A Visual Editor for Genetic Designs.
SBOLCanvas is a web-based application that can create and edit genetic constructs using the SBOL data and visual standards. SBOLCanvas allows a user to create a genetic design visually and structurally from start to finish. It also allows users to incorporate existing SBOL data from a SynBioHub repository. By the nature of being a web-based application, SBOLCanvas is readily accessible and easy to use. A live version of the latest release can be found at https://sbolcanvas.org.


A polymorphism of the GTP-cyclohydrolase I feedback regulator gene alters transcriptional activity and may affect response to SSRI antidepressants.
Tetrahydrobiopterin (BH(4)) is an essential cofactor for synthesis of many neurotransmitters including serotonin. In serotonergic neurons, BH(4) is tightly regulated by GTP-cyclohydrolase I feedback regulator (GFRP). Given the pivotal role of the serotonergic system in mood disorders and selective serotonin reuptake inhibitors (SSRIs) antidepressant function, we tested the hypothesis that GFRP gene (GCHFR) variants would modify response to antidepressants in subjects with major depression. Two single nucleotide polymorphisms (rs7164342 and rs7163862) in the GCHFR promoter were identified and occurred as two haplotypes (GA or TT). A multiple regression analysis revealed that homozygous individuals for the TT haplotype were less likely to respond to the SSRI fluoxetine than to the tricyclic antidepressant nortriptyline (P = 0.037). Moreover, the TT haplotype showed a reduced transcription rate in luciferase reporter gene assays, which may impact on BH(4)-mediated neurotransmitter production, thus suggesting a biological process through which GCHFR promoter variants might influence antidepressant response.


Ambient temperature and violent crime: tests of the linear and curvilinear hypotheses.
Laboratory research on the effects of temperature has led theorists to propose a curvilinear model relating negative affect and aggression. Two alternative explanations of these lab findings are proposed--one artifactual, one based on attributions for arousal. Both alternatives predict a linear relationship between temperature and aggression in real-world settings, whereas the negative affect curvilinear model predicts a specific curvilinear effect. Two studies are reported that investigated the relationship between temperature and violent crime. Both studies yielded significant linear relationships and failed to demonstrate the specified curvilinear relationship. Also, both studies yielded significant day-of-the-week effects. Implications of these findings for the study of aggression are discussed.


Beneath the label: unsatisfactory compliance with ESRB, PEGI and IARC industry self-regulation requiring loot box presence warning labels by video game companies.
Loot boxes in video games are a form of in-game transactions with randomized elements. Concerns have been raised about loot boxes' similarities with gambling and their potential harms (e.g. overspending). Recognizing players' and parents' concerns, in mid-2020, the Entertainment Software Rating Board (ESRB) and PEGI (Pan-European Game Information) announced that games containing loot boxes or any forms of in-game transactions with randomized elements will be marked by a new label stating 'In-Game Purchases (Includes Random Items)'. The same label has also been adopted by the International Age Rating Coalition (IARC) and thereby assigned to games available on digital storefronts, e.g. the Google Play Store. The label is intended to provide more information to consumers and allow them to make more informed purchasing decisions. This measure is not legally binding and has been adopted as industry self-regulation or corporate social responsibility. Previous research has suggested that industry self-regulation might not be effectively complied with due to conflicting commercial interests. Compliance with the ESRB's, PEGI's and IARC's loot box presence warning label was assessed in two studies. The first study found that 60.6% of all games labelled by either the ESRB or PEGI (or 16.1% using a more equitable methodology) were not labelled by the other. The majority of the inconsistencies were caused by the ESRB refusing to apply the measure retroactively. Five instances where one age rating organization culpably failed to accurately identify loot box presence were identified (although only two cases were admitted by the relevant organization). Generally, with newly released games, consumers can rely on the PEGI and ESRB labels. PEGI has retroactively labelled many older games, meaning that consumers can expect the labelling to be accurate. However, due to the ESRB's policies (which it has refused to improve), North American consumers cannot rely on the label for many older games containing loot boxes, unlike their European counterparts. The data suggest that the loot box issue is far more pressing on mobile platforms than console/PC platforms. The second study found that 71.0% of popular games containing loot boxes on the Google Play Store (whose age rating system is regulated through IARC) did not display the label and were therefore non-compliant. The IARC's current policy on the Google Play Store is that only games submitted for rating after February 2022 are required to be labelled. This policy (which the IARC has refused to improve) means that most popular and high-grossing games can be, and presently are, marketed without the label, thus significantly reducing the measure's scope and potential benefit. The Apple App Store still does not allow loot box presence to be disclosed. At present, consumers and parents cannot rely on this self-regulatory measure to provide accurate information as to loot box presence for mobile games. Due to their immense scale, the mobile markets pose regulatory and enforcement challenges that PEGI admits are not yet resolved. The mere existence of this measure cannot be used to justify the non-regulation of loot boxes by governments, given the poor compliance and doubtful efficacy (even if when complied with satisfactorily). Improvements to the existing age rating systems are proposed. Preregistered Stage 1 protocol: https://doi.org/10.17605/OSF.IO/E6QBM (date of in-principle acceptance: 12 January 2023).