Ecosystem biomonitoring with eDNA: metabarcoding across the tree of life in a tropical marine environment.
Effective marine management requires comprehensive data on the status of marine biodiversity. However, efficient methods that can document biodiversity in our oceans are currently lacking. Environmental DNA (eDNA) sourced from seawater offers a new avenue for investigating the biota in marine ecosystems. Here, we investigated the potential of eDNA to inform on the breadth of biodiversity present in a tropical marine environment. Directly sequencing eDNA from seawater using a shotgun approach resulted in only 0.34% of 22.3 million reads assigning to eukaryotes, highlighting the inefficiency of this method for assessing eukaryotic diversity. In contrast, using 'tree of life' (ToL) metabarcoding and 20-fold fewer sequencing reads, we could detect 287 families across the major divisions of eukaryotes. Our data also show that the best performing 'universal' PCR assay recovered only 44% of the eukaryotes identified across all assays, highlighting the need for multiple metabarcoding assays to catalogue biodiversity. Lastly, focusing on the fish genus Lethrinus, we recovered intra- and inter-specific haplotypes from seawater samples, illustrating that eDNA can be used to explore diversity beyond taxon identifications. Given the sensitivity and low cost of eDNA metabarcoding we advocate this approach be rapidly integrated into biomonitoring programs.


Asymmetric dimethylarginine correlates with measures of disease severity, major adverse cardiovascular events and all-cause mortality in patients with peripheral arterial disease.
Peripheral arterial disease (PAD) is associated with major cardiovascular morbidity and mortality. Abnormalities in nitric oxide metabolism due to excess of the NO synthase inhibitor asymmetric dimethylarginine (ADMA) may be pathogenic in PAD. We explored the association between ADMA levels and markers of atherosclerosis, function, and prognosis. A total of 133 patients with symptomatic PAD were enrolled. Ankle-brachial index (ABI), walking time, vascular function measures (arterial compliance and flow-mediated vasodilatation) and plasma ADMA level were assessed for each patient at baseline. ADMA correlated inversely with ABI (r = -0.238, p = 0.003) and walking time (r = -0.255, p = 0.001), independent of other vascular risk factors. We followed up 125 (94%) of our 133 initial subjects with baseline measurements (mean 35 months). Subjects with ADMA levels in the highest quartile (> 0.84 mumol/l) showed a significantly greater occurrence of a major adverse cardiovascular event (MACE) compared to those with ADMA levels in the lower three quartiles (p = 0.001). Cox proportional-hazards regression analysis revealed that ADMA was a significant predictor of MACE, independent of other risk factors including age, sex, blood pressure, smoking history, diabetes and ABI (hazard ratio = 5.1, p < 0.001). Measures of vascular function, such as compliance, flow-mediated vasodilatation (FMVD) and blood pressure, as well as markers of PAD severity, including ABI and walking time, were not predictive. In conclusion, circulating levels of ADMA correlate independently with measures of disease severity and major adverse cardiovascular events. Agents that target this pathway may be useful for this patient population. Clinical Trial Registration - URL: http:// www.clinicaltrials.gov. Unique identifier: NCT00284076.


Correlation of Symptoms, Clinical Signs, and Biomarkers of Inflammation in Postsurgical Chronic Rhinosinusitis.
The study aimed to evaluate symptoms described by patients with chronic rhinosinusitis with polypoid changes/nasal polyps and their correlation with computed tomography (CT), nasal endoscopy, and intranasal biomarkers. Prospective multicenter study symptom data from postsurgical adult chronic rhinosinusitis study participants with recurrent disease refractory to medical therapy were analyzed in comparison with objective data. Using logistic regression analysis, participant-rated 16-question surveys from 258 participants were assessed for correlation with nasal endoscopy scores, CT percentage of sinus occlusion, and intranasal biomarkers of fungal antigens (Alternaria and Aspergillus), eosinophilic inflammation (eosinophil-derived neurotoxin [EDN] and major basic protein [MBP]), and inflammatory cytokines (interleukins 5 and 13). Study participant assessments revealed increased CT occlusion in participants presenting with greater inability to smell ( P < .019). Mucosal inflammation identified on nasal endoscopy was positively correlated with congestion ( P < .028), runny nose ( P < .002), and ear pain ( P < .007). Elevated EDN was positively correlated in patients with bothersome congestion ( P < .031) and runny nose ( P < .011). Sneezing was positively correlated with multiple markers: Alternaria ( P < .024), interleukin-13 ( P < .027), MBP ( P < .034), and interleukin-5 ( P < .019). Nasal endoscopy, not CT imaging, has the strongest correlation with the 2 cardinal symptoms of congestion and runny nose in CRS patients; these correlate with biomarkers of eosinophilic inflammation.


The economic burden of households affected by tuberculosis in Brazil: First national survey results, 2019-2021.
One of the three main targets of the World Health Organization (WHO) End TB Strategy (2015-2035) is that no tuberculosis (TB) patients or their households face catastrophic costs (defined as exceeding 20% of the annual household income) because of the disease. Our study seeks to determine, as a baseline, the magnitude and main drivers of the costs associated with TB disease for patients and their households and to monitor the proportion of households experiencing catastrophic costs in Brazil. A national cross-sectional cluster-based survey was conducted in Brazil in 2019-2021 following WHO methodology. TB patients of all ages and types of TB were eligible for the survey. Adult TB patients and guardians of minors (<18 years old) were interviewed once about costs, time loss, coping measures, income, household expenses, and asset ownership. Total costs, including indirect costs measured as reported household income change, were expressed as a percentage of annual household income. We used descriptive statistics to analyze the cost drivers and multivariate logistic regression to determine factors associated with catastrophic costs. We interviewed 603 patients, including 538 (89%) with drug-sensitive (DS) and 65 (11%) with drug-resistant (DR) TB. Of 603 affected households, 48.1% (95%CI: 43-53.2) experienced costs above 20% of their annual household income during their TB episode. The proportion was 44.4% and 78.5% among patients with DS- and DR-TB, respectively. On average, patients incurred costs of US$1573 (95%CI: 1361.8-1785.0) per TB episode, including pre-diagnosis and post-diagnosis expenses. Key cost drivers were post-diagnosis nutritional supplements (US$317.6, 95%CI: 232.7-402.6) followed by medical costs (US$85.5, 95%CI: 54.3-116.5) and costs of travel for clinic visits during treatment (US$79.2, 95%CI: 61.9-96.5). In multivariate analysis, predictors of catastrophic costs included positive HIV status (aOR = 3.0, 95%CI:1.1-8.6) and self-employment (aOR = 2.7, 95%CI:1.1-6.5); high education was a protective factor (aOR = 0.1, 95%CI:0.0-0.9). Although the services offered to patients with TB are free of charge in the Brazilian public health sector, the availability of free diagnosis and treatment services does not alleviate patients' financial burden related to accessing TB care. The study allowed us to identify the costs incurred by patients and suggest actions to mitigate their suffering. In addition, this study established a baseline for monitoring catastrophic costs and fostering a national policy to reduce the costs to patients for TB care in Brazil.


[Immunological detection of sphingomyelin exchange protein in tumors and in embryonic rat liver].
The interaction of postmicrosomal supernatants from the Jensen sarcoma, sarcoma 45, nephroma RA, normal, regeherating and fetal rat livers with antisera to the sphingomyelin exchange protein isolated from hepatoma 27 was studied by double immunodiffusion. It was shown that this protein is present in comparable amounts in all the tumours tested and in fetal rat liver, whereas in normal and regenerating liver its content is negligible. Based on the correlation between the content of sphingomyelin exchange protein in cell cytosols and the amount of sphingomyelin in the corresponding mitochondria it is suggested that this protein is responsible for the presence of sphingomyelin in mitochondria.


Open Versus Closed Group Treatment of Men with a History of Sexual Offenses.
Treatment programs for people with histories of sexual violence form a critical part of criminal justice service rehabilitation. Completion of these programs is often a precondition of release. Meta-analytic reviews suggest moderate benefit is associated with treatment completion, although effect sizes vary. This study examined whether commencement of open versus closed group programs was associated with treatment completion and recidivism. Participants were 426 adult men who commenced treatment between April 1, 2014, and December 31, 2017. Participants were followed-up until June 30, 2018. Programs varied by type (open versus closed), location (in-custody versus in-community) and intensity (moderate versus high). No significant differences were observed between open and closed programs for treatment completion but men who were treated in-custody were more likely to complete treatment when compared to those men who commenced treatment in the community. No significant differences were observed between open and closed programs for sexual or for any recidivism.


AntagomiR-451 inhibits oxygen glucose deprivation (OGD)-induced HUVEC necrosis via activating AMPK signaling.
Oxygen glucose deprivation (OGD) application in cultured human umbilical vein endothelial cells (HUVECs) mimics ischemic injuries. AntagomiR-451, the miroRNA-451 ("miR-451") inhibitor, could activate pro-survival AMP-activated protein kinase (AMPK) signaling. In the current study, we showed that forced-expression of antagomiR-451 depleted miRNA-451 and significantly attenuated OGD-induced necrosis of HUVECs. Activation of AMPK was required for antagomiR-451-mediated pro-survival actions. AMPK inhibition, by AMPKα shRNA or dominant negative mutation, almost completely abolishedantagomiR-451-mediated HUVEC protection again OGD. Reversely, forced-activation of AMPK by exogenous expression of constructively-active AMPKα inhibited OGD-induced HUVEC necrosis. At the molecular level, antagomiR-451 expression in HUVECs inhibited OGD-induced programmed necrosis, the latter was evidenced by mitochondrial p53-cyclophilinD (Cyp-D) association, mitochondrial depolarization as well as reactive oxygen species (ROS) production and lactate dehydrogenase (LDH) breach. Together, we suggest that antagomiR-451 activates AMPK to inhibit OGD-induced programmed necrosis in HUVECs.


Invited review: Genetics and claw health: Opportunities to enhance claw health by genetic selection.
Routine recording of claw health status at claw trimming of dairy cattle has been established in several countries, providing valuable data for genetic evaluation. In this review, we examine issues related to genetic evaluation of claw health; discuss data sources, trait definitions, and data validation procedures; and present a review of genetic parameters, possible indicator traits, and status of genetic and genomic evaluations for claw disorders. Different sources of data and traits can be used to describe claw health. Severe cases of claw disorders can be identified by veterinary diagnoses. Data from lameness and locomotion scoring, activity information from sensors, and feet and leg conformation traits are used as auxiliary traits. The most reliable and comprehensive information is data from regular hoof trimming. In genetic evaluation, claw disorders are usually defined as binary traits, based on whether or not the claw disorder was present (recorded) at least once during a defined time period. The traits can be specific disorders, composite traits, or overall claw health. Data validation and editing criteria are needed to ensure reliable data at the trimmer, herd, animal, and record levels. Different strategies have been chosen, reflecting differences in herd sizes, data structures, management practices, and recording systems among countries. Heritabilities of the most commonly analyzed claw disorders based on data from routine claw trimming were generally low, with ranges of linear model estimates from 0.01 to 0.14, and threshold model estimates from 0.06 to 0.39. Estimated genetic correlations among claw disorders varied from -0.40 to 0.98. The strongest genetic correlations were found among sole hemorrhage (SH), sole ulcer (SU), and white line disease (WL), and between digital/interdigital dermatitis (DD/ID) and heel horn erosion (HHE). Genetic correlations between DD/ID and HHE on the one hand and SH, SU, or WL on the other hand were, in most cases, low. Although some of the studies were based on relatively few records and the estimated genetic parameters had large standard errors, there was, with some exceptions, consistency among studies. Various studies evaluate the potential of various data soureces for use in breeding. The use of hoof trimming data is recommended for maximization of genetic gain, although auxiliary traits, such as locomotion score and some conformation traits, may be valuable for increasing the reliability of genetic evaluations. Routine genetic evaluation of direct claw health has been implemented in the Netherlands (2010); Denmark, Finland, and Sweden (joint Nordic evaluation; 2011); and Norway (2014), and other countries plan to implement evaluations in the near future.


Developmental DNA rearrangements and micronucleus-specific sequences in five species within the Tetrahymena pyriformis species complex.
In Tetrahymena thermophila, the development of a transcriptionally active macronucleus from a transcriptionally inert micronucleus includes the elimination of many segments of DNA, the bulk of which belong to repetitive sequence families. Two approaches were used to study the interspecies variations in developmentally eliminated DNA segments. First, the occurrence of restriction fragments crosshybridizing to developmentally eliminated DNA segments isolated from T. thermophila was examined in other species of Tetrahymena. Most micronucleus-specific sequence families examined showed large differences in numbers and intensities of crosshybridizing bands in different species, indicating the possibility of gain or loss of repeats within each of the sequence families. Second, the presence of developmentally excisable DNA segments, i.e., of rearrangement sites, was examined in the same set of species at a number of unique loci. This was carried out by comparing the hybridization patterns of seven unique macronucleus-retained sequences in the micro- and macronuclei of each of the species. Essentially all of the loci displayed variability with respect to the presence of rearrangement sites among the species examined. Results from the two approaches indicate that generation or loss of developmental rearrangements can occur among the species examined here.


Estimating molar-incisor-hypomineralization among 8-year-olds based on 15-year public oral health practice-based data.
Background: A wide range in the prevalence of molar-incisor-hypomineralization (MIH) has been reported. Population-based studies are recommended. However, such studies are expensive and time-consuming.Objectives: To estimate the magnitude of MIH condition among 8-year-olds based on routine oral health examinations and to associate first permanent molar (FPM) affection with that of other permanent teeth over time.Materials and methods: This retrospective study, with cross-sectional and longitudinal components, was based on electronic oral health records; all 8-year-olds examined between 2002 and 2016 were included.Results: The average estimated prevalence of MIH was 8.3%; yearly range was 4.8-15.9%. The mean number of affected teeth was 1.4; 62% had one affected tooth. One-surface defects were the most frequent (66%). Asymmetric distribution of affected teeth was observed. In follow-up, 10.7%, 8.4%, and 11.2% had at least one affected permanent canine, premolar, or second permanent molar, respectively. The proportion of children with other MIH-affected permanent teeth was higher in the group with ≥2 MIH-affected teeth than in group with one affected FPM at the age of 8.Conclusions: The average prevalence of MIH was comparable to that reported elsewhere. The number of MIH-affected teeth at early mixed dentition predicts the affection of other permanent teeth over time, mainly that of permanent canines. Further screening of children with MIH is recommended to improve individually tailored early preventive and restorative dental care.


Multicenter, randomized, comparative cost-effectiveness study comparing 0, 1, and 2 diagnostic medial branch (facet joint nerve) block treatment paradigms before lumbar facet radiofrequency denervation.
Among patients presenting with axial low back pain, facet arthropathy accounts for approximately 10-15% of cases. Facet interventions are the second most frequently performed procedures in pain clinics across the United States. Currently, there are no uniformly accepted criteria regarding how best to select patients for radiofrequency denervation. A randomized, multicenter study was performed in 151 subjects with suspected lumbar facetogenic pain comparing three treatment paradigms. Group 0 received radiofrequency denervation based solely on clinical findings; group 1 underwent denervation contingent on a positive response to a single diagnostic block; and group 2 proceeded to denervation only if they obtained a positive response to comparative blocks done with lidocaine and bupivacaine. A positive outcome was predesignated as > or =50% pain relief coupled with a positive global perceived effect persisting for 3 months. In group 0, 17 patients (33%) obtained a successful outcome at 3 months versus eight patients (16%) in group 1 and 11 (22%) patients in group 2. Denervation success rates in groups 0, 1, and 2 were 33, 39, and 64%, respectively. Pain scores and functional capacity were significantly lower at 3 months but not at 1 month in group 2 subjects who proceeded to denervation compared with patients in groups 0 and 1. The costs per successful treatment in groups 0, 1, and 2 were $6,286, $17,142, and $15,241, respectively. Using current reimbursement scales, these findings suggest that proceeding to radiofrequency denervation without a diagnostic block is the most cost-effective treatment paradigm.


Root contraction in Cycas and Zamia (Cycadales) determined by gelatinous fibers.
• Reaction wood (RW) in seed plants can induce late and usually secondary changes in organ orientation. Conifers produce compression wood (CW), generated by compression tracheids, which generate a push force. Angiosperms produce tension wood (TW), generated by tension wood fibers (TWF) often described as "gelatinous fibers," which exert a pull force. Usually RW is produced eccentrically, but it can occur concentrically, as in aerial roots of Ficus. However, gymnosperms can produce gelatinous fibers (tension fibers, TF), as in cortical and secondary phloem tissues (Gnetum). TFs are therefore limited neither to wood, xylem, nor angiosperms. Here we demonstrate that TFs in secondary phloem are involved in contraction of roots of cycads and compare them with TFs of Ficus.• We sectioned root material of cycads at various stages of seedling development using simple staining and histochemical procedures to follow the course of secondary phloem development. Aerial roots of Ficus were compared with the cycad root material.• Tension fibers (gelatinous fibers) occur extensively and continuously in the secondary phloem in roots that undergo contraction. Older tissues, but notably the xylem, become distorted by contraction. TFs in cycads correspond in cell wall features to TFs that occur in Ficus, but do not occur in secondary xylem. The individual fibers visibly contract.• Tissue contraction in Cycas and Zamia corresponds to that found in angiosperms and Gnetum and further broadens the scope of the activity of tension tissues. This finding possibly indicates that gelatinous fibers originated at a very early period of seed plant evolution.


Thalamocortical abnormalities in auditory brainstem response patterns distinguish DSM-IV bipolar disorder type I from schizophrenia.
Bipolar disorder type I (BP-I) belongs to a spectrum of affective disorders that are expressed in many different ways and therefore can be difficult to distinguish from other conditions, especially unipolar depression, schizoaffective disorder, schizophrenia (SZ), but also anxiety and personality disorders. Since early diagnosis and treatment have shown to improve the long-term prognosis, complementary specific biomarkers are of great value. The auditory brainstem response (ABR) has previously been applied successfully to identify specific abnormal ABR patterns in SZ and Asperger syndrome. The current study investigated the early auditory processing of complex sound stimuli e.g. forward masking, in BP-I compared to SZ patients. The ABR curves of BP-I patients (n=23) and SZ patients (n=20) were analyzed in terms of peak amplitudes and correlation with an ABR norm curve based on a non-psychiatric control group (n=20). BP-I patients had significantly higher wave III (p=0.0062) and wave VII (p=0.0472) amplitudes compared with SZ patients. Furthermore, BP-I patients, and to a lesser extent SZ patients, showed low correlation with the norm ABR curve in the part of the curve comprising waves VI-VII. Sample size was relatively small and study groups were not matched for age and gender. BP-I patients showed specific aberrances, specifically in the latter part of the ABR curve, implicating abnormalities in thalamocortical circuitry. The abnormal ABR wave patterns significantly separated BP-I patients from SZ patients suggesting that ABR might serve as a biomarker for BP-I.


Production of monoclonal antibodies for the immunohistochemical detection of gastric carcinomas.
About 700 antibody-secreting hybrids were obtained by fusion of lymphocytes, (harvested from mice hyperimmunized with the human gastric carcinoma line KATO III) and P3-X63-Ag8-653 myeloma cells. Antibody specificity was screened in ELISA performed on glutaraldehyde-fixed cultured cells and on paraffin-embedded tissue sections stained with the method of avidin-biotin-peroxidase. When tested in ELISA, the monoclonal antibody produced by the hybrid clone BD-5 was found to bind only to the cell line used as immunogen, among the many neoplastic or normal human cell lines tested. When assayed on paraffin sections with the avidin-biotin-peroxidase method, the BD-5 monoclonal antibody stained gastric carcinomas, but not the normal mucosa. Pancreatic carcinomas were also stained, while the corresponding normal gland was not. The antibody strongly stained the normal colonic and small intestinal mucosa. Among the other normal or neoplastic tissues tested, a weak reactivity was observed only with some epithelial cells of the salivary glands and with some carcinoma cells of the uterus and of the lung. It is concluded that the BD-5 antibody reacts with an epitope normally present on intestinal mucosa, which, following neoplastic transformation, is ectopically expressed also on gastric and pancreatic carcinomas. This monoclonal could represent a useful reagent for histopathological diagnosis.


Functional rescue of elastin insufficiency in mice by the human elastin gene: implications for mouse models of human disease.
Diseases linked to the elastin gene arise from loss-of-function mutations leading to protein insufficiency (supravalvular aortic stenosis) or from missense mutations that alter the properties of the elastin protein (dominant cutis laxa). Modeling these diseases in mice is problematic because of structural differences between the human and mouse genes. To address this problem, we developed a humanized elastin mouse with elastin production being controlled by the human elastin gene in a bacterial artificial chromosome. The temporal and spatial expression pattern of the human transgene mirrors the endogenous murine gene, and the human gene accurately recapitulates the alternative-splicing pattern found in humans. Human elastin protein interacts with mouse elastin to form functional elastic fibers and when expressed in the elastin haploinsufficient background reverses the hypertension and cardiovascular changes associated with that phenotype. Elastin from the human transgene also rescues the perinatal lethality associated with the null phenotype. The results of this study confirm that reestablishing normal elastin levels is a logical objective for treating diseases of elastin insufficiency such as supravalvular aortic stenosis. This study also illustrates how differences in gene structure and alternative splicing present unique problems for modeling human diseases in mice.


PAX3 is expressed in human melanomas and contributes to tumor cell survival.
Deregulated expression of the transcription factor PAX3 was observed previously in several tumors like rhabdomyosarcoma and Ewing's sarcoma. Because PAX3 expression is also found in pluripotent neural crest cells, we investigated whether melanomas, tumors derived mostly from cutaneous intraepidermal melanocytes, might show deregulated PAX3 expression. Using a specific and sensitive reverse transcription-PCR, we detected PAX3 mRNA in 77% (27 of 35) of primary cultured melanomas. These results could be confirmed by direct in situ hybridization on the corresponding tissue sections where PAX3 expression was unambiguously confined to tumor cells and not detected in surrounding normal tissue, normal skin sections, or sections of benign lesions. Furthermore, down-regulation of PAX3 expression achieved through a specific antisense oligonucleotide-based treatment resulted in > 70% of dead cells specifically in PAX3-positive melanomas. Annexin V staining confirmed that primary melanoma cells underwent apoptosis after treatment These experiments suggest that in situ hybridization of PAX3 on paraffin-embedded tissue may represent a novel means to identify melanoma cell lesions, which appear to become dependent on expression of this deregulated transcription factor.


Immune biomarkers and anti-HIV activity in the reproductive tract of sexually active and sexually inactive adolescent girls.
Adolescent girls are disproportionately affected by the HIV/AIDS pandemic, accounting for 22% of all new HIV infections globally. Yet little is known regarding the immune microenvironment of the adolescent female reproductive tract, especially regarding differences among sexually active and inactive girls, a critical parameter to evaluate HIV susceptibility associated with young age and sexual debut. Cervico-vaginal lavage (CVL) was collected from sexually active (10) and inactive (8) girls aged 11-19 years and analyzed by ELISA for inflammation-associated biomarkers IL-6, IL-8, TNF-α, MIP-3α, IL-1α, IL-1β, matrix metalloproteinases (MMP) 1, 2, 7, 8, and 9, as well as anti-HIV mediators, Elafin, SLPI, human beta-defensin 2, and tissue inhibitor of matrix metalloproteinases (TIMP) 1 and 2. Cervical ectopy was analyzed using Volocity. Anti-HIV activity was determined by TZM-bl assay. Statistical analyses were performed using GraphPad Prism and R. Sexually inactive girls had significantly higher levels of TNF-α (P = .029) in CVL compared to sexually active girls. In contrast, sexually active girls showed a trend toward higher levels of IL-1α (P = .051) compared to the sexually inactive girls. Heat-map correlations between cervical ectopy and immune biomarkers were also distinct between the 2 populations with significant positive associations between % ectopy and inflammation-associated biomarkers IL-6, IL-1β, IL-8, MIP-3α, MMP-8, and MMP-9 observed in the sexually inactive but not sexually active group. Higher pro-inflammatory biomarker TNF-α, as well as a distinct inflammation-associated immune clustering in sexually inactive girls, can potentially increase risk for infections including HIV upon sexual debut. Future studies with larger sample sizes are needed to characterize the immune parameters associated with sexual activity.


Neuroprotection by IFN-γ via astrocyte-secreted IL-6 in acute neuroinflammation.
Inflammation eliminates pathogenic infections while also threatening the integrity of the central nervous system. In this study, using in vivo and in vitro models of acute neuroinflammation, we investigated the mechanisms by which inflammation and astrocytes affect neuronal apoptosis. The in vitro model mimicked acute neuroinflammation by incubation in IFN-γ-containing media with primary cultured cerebellar granule neurons, with or without cultured astrocytes. This quickly induced neuronal apoptosis characterized by cleaved caspase-3 expression, Hoechst 33342 staining, and intercellular Ca2+ influx, whereas the presence of astrocytes significantly protected neurons from these effects. IFN-γ in the inflammation media also promoted astrocyte secretion of IL-6, essential for protection. The supernatants of rat peripheral blood mononuclear cells stimulated by lymphocyte mitogen lipopolysaccharide or concanavalin A were used as inflammation media to verify the results. The in vivo model involved a peripheral challenge with lipopolysaccharide, with or without recombinant IFN-γ, in C57BL/6 mice. This confirmed the in vitro results: anti-IFN-γ antibodies exacerbated the acute course of neuroinflammation and led to neurocyte apoptosis in vivo. The pro-inflammatory cytokine IFN-γ provided neuroprotection during acute neuroinflammation via induction of astrocyte-secreted IL-6. The findings provide novel insights into the mechanisms of neuroprotection by IFN-γ during acute neuroinflammation, and may impact therapies for inflammation-related central nervous system injury and disease.


Acceptability of and willingness to pay for using a smartphone-based vaccination application in a Vietnamese cohort.
The increasing accessibility of smartphone in Vietnam shows potential in using smartphone applications for vaccination management to improve compliance. However, the acceptability and financial feasibility of using this application in Vietnam have not yet been understood. This study measured the general perception of and willingness to pay (WTP) for using smartphone-based vaccination management applications and their associated factors in Vietnam. A cross-sectional study was conducted between March and April 2016 in an urban vaccination clinic at the Hanoi Medical University in Vietnam. Convenience sampling was used to recruit 429 adult participants who had received vaccinations or whose children were vaccinated. Sociodemographic characteristics and the acceptability of and WTP for using smartphone-based vaccination management applications were evaluated. Among participants who used smartphone applications, 5% were aware of existing vaccination management applications, of whom 57.9% had previously used the applications. About 69.6% of participants believed that the applications were necessary, 93.7% of them were also willing to use the applications, and 79.1% were willing to pay 92.7 thousand Vietnamese dong (VND) for the applications on average. Participants who were older, unemployed, earned more household income, and having knowledge about functions and benefits of vaccination were less likely to use the vaccination applications. Participants who brought their children to get vaccinated at younger age and with higher level of education were willing to pay more for vaccination applications. High levels of acceptability of, willingness to use, and WTP for smartphone-based vaccination management applications among Vietnamese participants are reported. Increased education and awareness about the benefits of vaccination and vaccination management applications through community health workers might increase the feasibility of implementing such applications and perception toward their usage among young adult smartphone users. In addition, building a stronger relationship with health care providers at hospitals might improve the application's visibility and adoption.


PERFect: PERmutation Filtering test for microbiome data.
The human microbiota composition is associated with a number of diseases including obesity, inflammatory bowel disease, and bacterial vaginosis. Thus, microbiome research has the potential to reshape clinical and therapeutic approaches. However, raw microbiome count data require careful pre-processing steps that take into account both the sparsity of counts and the large number of taxa that are being measured. Filtering is defined as removing taxa that are present in a small number of samples and have small counts in the samples where they are observed. Despite progress in the number and quality of filtering approaches, there is no consensus on filtering standards and quality assessment. This can adversely affect downstream analyses and reproducibility of results across platforms and software. We introduce PERFect, a novel permutation filtering approach designed to address two unsolved problems in microbiome data processing: (i) define and quantify loss due to filtering by implementing thresholds and (ii) introduce and evaluate a permutation test for filtering loss to provide a measure of excessive filtering. Methods are assessed on three "mock experiment" data sets, where the true taxa compositions are known, and are applied to two publicly available real microbiome data sets. The method correctly removes contaminant taxa in "mock" data sets, quantifies and visualizes the corresponding filtering loss, providing a uniform data-driven filtering criteria for real microbiome data sets. In real data analyses PERFect tends to remove more taxa than existing approaches; this likely happens because the method is based on an explicit loss function, uses statistically principled testing, and takes into account correlation between taxa. The PERFect software is freely available at https://github.com/katiasmirn/PERFect.


Does Metformin Reduce Cancer Risks? Methodologic Considerations.
The substantial burden of cancer and diabetes and the association between the two conditions has been a motivation for researchers to look for targeted strategies that can simultaneously affect both diseases and reduce their overlapping burden. In the absence of randomized clinical trials, researchers have taken advantage of the availability and richness of administrative databases and electronic medical records to investigate the effects of drugs on cancer risk among diabetic individuals. The majority of these studies suggest that metformin could potentially reduce cancer risk. However, the validity of this purported reduction in cancer risk is limited by several methodological flaws either in the study design or in the analysis. Whether metformin use decreases cancer risk relies heavily on the availability of valid data sources with complete information on confounders, accurate assessment of drug use, appropriate study design, and robust analytical techniques. The majority of the observational studies assessing the association between metformin and cancer risk suffer from methodological shortcomings and efforts to address these issues have been incomplete. Future investigations on the association between metformin and cancer risk should clearly address the methodological issues due to confounding by indication, prevalent user bias, and time-related biases. Although the proposed strategies do not guarantee a bias-free estimate for the association between metformin and cancer, they will reduce synthesis of and reporting of erroneous results.


Lentiviral vector delivery of human interleukin-7 (hIL-7) to human immune system (HIS) mice expands T lymphocyte populations.
Genetically modified mice carrying engrafted human tissues provide useful models to study human cell biology in physiologically relevant contexts. However, there remain several obstacles limiting the compatibility of human cells within their mouse hosts. Among these is inadequate cross-reactvitiy between certain mouse cytokines and human cellular receptors, depriving the graft of important survival and growth signals. To circumvent this problem, we utilized a lentivirus-based delivery system to express physiologically relevant levels of human interleukin-7 (hIL-7) in Rag2-/-gammac-/- mice following a single intravenous injection. hIL-7 promoted homeostatic proliferation of both adoptively transferred and endogenously generated T-cells in Rag2-/-gammac-/- Human Immune System (HIS) mice. Interestingly, we found that hIL-7 increased T lymphocyte numbers in the spleens of HIV infected HIS mice without affecting viral load. Taken together, our study unveils a versatile approach to deliver human cytokines to HIS mice, to both improve engraftment and determine the impact of cytokines on human diseases.


Effect of drugs on urate binding to plasma proteins.
The effect of various drugs on urate binding to plasma proteins was investigated in normal subjects. Whereas allopurinol, aspirin, phenylbutazone, probenecid, and sulphinpyrazone all significantly reduced plasma urate concentrations, only aspirin, phenylbutazone, and probenecid significantly impaired urate binding. Colchicine and indomethacin in the doses administered had no significant effect on plasma urate concentrations or binding. In the case of aspirin, urate binding was reduced to 25% of normal, and this effect was quickly abolished after cessation of therapy. Phenylbutazone reduced urate binding to 56% and probenecid to 46% of normal; this impairment was still detected four days after cessation of therapy. Drugs may impair urate binding by competition for plasma protein binding sites, with displacement of bound urate. Impairment of urate binding in vivo by administration of certain drugs may be relevant to the precipitation of acute gouty arthritis, to the formation of gouty tophi, and to the augmentation of uricosuria. Furthermore, the role of drugs must be seriously considered during all studies on urate binding in patients with gout.


Sonosensitive Cavitation Nuclei-A Customisable Platform Technology for Enhanced Therapeutic Delivery.
Ultrasound-mediated cavitation shows great promise for improving targeted drug delivery across a range of clinical applications. Cavitation nuclei-sound-sensitive constructs that enhance cavitation activity at lower pressures-have become a powerful adjuvant to ultrasound-based treatments, and more recently emerged as a drug delivery vehicle in their own right. The unique combination of physical, biological, and chemical effects that occur around these structures, as well as their varied compositions and morphologies, make cavitation nuclei an attractive platform for creating delivery systems tuned to particular therapeutics. In this review, we describe the structure and function of cavitation nuclei, approaches to their functionalization and customization, various clinical applications, progress toward real-world translation, and future directions for the field.


Effect of chromophore encapsulation on linear and nonlinear optical properties: the case of "miniSOG", a protein-encased flavin.
Linear and nonlinear spectroscopic parameters of flavin mononucleotide, FMN, have been examined both experimentally and computationally under conditions in which FMN is (1) solvated in a buffered aqueous solution, and (2) encased in a protein that is likewise solvated in a buffered aqueous solution. The latter was achieved using "miniSOG" which is an FMN-containing protein engineered from Arabidopsis thaliana phototropin 2. Although it is reasonable to expect that the encasing protein could have an appreciable effect, certainly on the nonlinear two-photon absorption cross section, we find that replacing the dynamic aqueous environment with the more static protein environment does little to influence the spectroscopic properties of FMN. The experimental and computational studies are consistent in this regard, and this agreement indicates that comparatively high-level computational methods can indeed be used with success on large chromophores with a complicated local environment. The results of the present study facilitate the much-needed development of well-characterized and readily-controlled chromophores suitable for use as intracellular sensitizers and fluorophores.


Liver cell uptake and degradation of soluble immunoglobulin G immune complexes in vivo and in vitro in rats.
Immune complexes were formed between dinitrophenylated human serum albumin (DNP-HSA) and polyclonal rabbit immunoglobulin G (IgG) anti-DNP antibodies at antibody excess. The antigen was labelled with isotope (125I-tyramine-cellobiose) or fluorochrome, (6-[fluorescein-5-(and-6)-carboxamido] hexanoic-acid, succinimidyl ester). The radiolabelled antigen, native or antibody complexed, was given intravenously to rats. Radioactivity was measured in various organs at 1 hour following injection. The liver was the main site for removal of the antigen as well as of the immune complexes. Within the liver, immune complexes were mainly associated with nonparenchymal liver cells, the total recovery from Kupffer cells being about 10 times greater than from the liver endothelial cells. The uncomplexed radiolabelled antigen was readily degraded by both cells types. After IgG complexing, the degradation decreased, both in Kupffer cells and in liver endothelial cells. In vitro experiments with isolated liver cells, showed that IgG complexing increased antigen uptake to about the same extent in Kupffer cells and in liver endothelial cells. The degradation of both antigen and immune complexes was less efficient in vitro than in vivo. Immune complex uptake in vitro was shown also by confocal fluorescence microscopy in Kupffer cells and in liver endothelial cells. Also in vitro, only minor uptake was found in the hepatocytes. We conclude that both liver endothelial cells and Kupffer cells are involved in the hepatic handling of soluble IgG immune complexes, but we found no evidence for substantial uptake by hepatocytes.


Cost-effectiveness of polatuzumab vedotin in relapsed or refractory diffuse large B-cell lymphoma.
A recent phase II trial showed that use of polatuzumab vedotin in combination with bendamustine plus rituximab (Pola-BR) in transplant-ineligible patients with relapsed or refractory (R/R) diffuse large B-cell lymphoma (DLBCL) resulted in longer progression-free survival and overall survival compared to bendamustine plus rituximab (BR) alone. In this study, we constructed a Markov model to assess the cost-effectiveness of Pola-BR versus BR in transplant-ineligible R/R DLBCL. We calculated the incremental cost-effectiveness ratio (ICER) of each treatment strategy from a US payer perspective, using a lifetime horizon and a willingness-to-pay threshold of $100,000 per quality-adjusted life-year (QALY). Use of Pola-BR was associated with an incremental cost of $92,641 compared to BR alone ($200,905 vs $108,265, respectively), an incremental effectiveness of 1.76 QALYs (2.35 vs 0.59 QALYs, respectively), and an ICER of $52,519/QALY. These data suggest that use of Pola-BR for R/R DLBCL is likely to be cost-effective compared to BR alone.


Correlation of dermoscopic structures of melanocytic lesions to reflectance confocal microscopy.
To determine the utility of reflectance confocal microscopy (RCM) in the in vivo evaluation of dermoscopic structures of melanocytic lesions. For each described dermoscopic feature, we evaluated by RCM at least 2 melanocytic lesions. A digital camera connected to the confocal computer enabled direct analysis of the dermoscopic structures. To ascertain precision of correlation, the orientation of the dermoscopic and RCM images were compared using a superimposed grid. Dermatology clinic specializing in pigmented lesions. Patients Eleven patients with melanocytic lesions, including 2 melanomas, 1 Spitz nevus, 7 dysplastic nevi, and 1 compound nevus. Main Outcome Measure Direct correlation of structures seen using dermoscopy with those seen using RCM. There was a good correlation between the global dermoscopic pattern and findings on the 4 x 4-mm mosaic of confocal images at the level of the dermoepidermal junction. The atypical network correlated with variability in the size and shape of dermal papillae. Globules corresponded with aggregates of bright cells, and darker shades of brown on dermoscopy appeared brighter on RCM. In peripheral streaks, RCM showed dense aggregates of pleomorphic cells of variable brightness and ill-defined cellular borders. These aggregates were continuous with the bright mesh that composed the central bulk of the lesion. A blue-white veil correlated with disruption of the rimmed papillae meshlike pattern and sometimes with the presence of bright cells corresponding to melanophages. Correlating dermoscopic structures to RCM features is possible and a necessary step toward understanding the potential benefits of RCM in the clinical setting.


Estimation of Duloxetine Hydrochloride in Pharmaceutical Formulations by RP-HPLC Method.
Simple, specific, accurate and precise method, namely, reverse phase high performance liquid chromatography was developed for estimation of duloxetine HCl in pharmaceutical formulations. For the high performance liquid chromatography method, Phenomenox C-18, 5 μm column consisting of 250×4.6 mm i.d. in isocratic mode, with mobile phase containing 0.01M 5.5 pH phosphate buffer: acetonitrile (60:40 v/v) and final pH adjust to 5.5±0.02 with phosphoric acid was used. The flow rate was 1.2 ml/min and effluent was monitored at 231 nm. The retention time was 5.61 min. The method was validated in terms of linearity, accuracy and precision. The linearity curve was found to be linear over 0.25-4 μg/ml. The limit of detection and limit of quantification were found to be 0.10 and 0.25 μg/ml respectively. The proposed method was successfully used to determine the drug content of marketed formulations.


Age differences and developmental trends in alarm peep responses by squirrel monkeys (Saimiri sciureus).
Alarm calls can code for different classes of predators or different types of predatory threat. Acoustic information can also encode the urgency of threat through variations in acoustic features within specific alarm call types. Squirrel monkeys (Saimiri sciureus) produce an alarm call, known as the alarm peep, in highly threatening situations. Infant squirrel monkeys appear to have an innate predisposition to respond to alarm peeps but require experience to associate alarm peeps with the appropriate type of predatory threat [Herzog & Hopf, American Journal of Primatology 7:99-106, 1984]. Little is known about age-related differences in the type or frequency of response to alarm peeps, or the development of alarm peep response in infants. The purpose of this study was to test experimentally the response strategies of different age classes of squirrel monkey to the playback of alarm peeps that were produced by infants, juveniles, or adults. Results suggest that infants, juveniles, and female subadults respond more frequently to alarm peeps than do adult females. Infant squirrel monkeys showed different behavioral strategies in response to alarm peeps as a function of age. Adult females differentiate between infant and adult alarm peeps by responding more frequently to the alarm peeps of adult females. These data demonstrate that squirrel monkeys use acoustic information to discern when to respond to the alarm peeps from conspecifics, and that infants gradually develop an adult-like response to alarm peeps over the first year of development.


Steady-state effects of extracellular potassium concentration on vascular smooth muscle reactivity.
Helical strips cut from small arteries taken from rabbit skeletal muscle were used to determine the steady-state effects of extracellular potassium concentration ([K]o) on the reactivity to epinephrine or norepinephrine. It was found that reactivity observed under the usual steady-state conditions (termed "usual reactivity") was depressed as steady-state [K]o was reduced from 8 to 1 mM. This depression reflected concurrent changes in two components of contraction: 1) a positive component (evaluated as reactivity during brief exposure to 10(-6) M ouabain) decreased when steady-state [K]o was reduced below approximately 5 mM, and 2) a negative component (evaluated indirectly as enhancement of reactivity produced by 10(-6) M ouabain) increased to a peak when steady-state [K]o was reduced from 8 to approximately 4-5 mM and decreased below this peak when steady-state [K]o was reduced below approximately 4-5 mM. It was also found that the magnitude of a reserve ouabain-sensitive negative component (demonstrated by abruptly increasing [K]o to 10.18 mM) increased from a minimum to a maximum level when steady-state [K]o was reduced from 6 to 3 mM. It is suggested that the effects of [K]o on the positive component of contraction, defined earlier as contractility, primarily involve some factor(s) other than transmembrane potential, whereas the effects on the negative component involve transmembrane potential exclusively and, in particular, the electrogenic potential. The latter suggestion constitutes the basis for use of the term electrogenesis in reference to this negative component.


Neonatal morbidities and developmental delay in moderately preterm-born children.
Children born moderately preterm (32-35(6/7) weeks' gestation) are at increased risk of both neonatal morbidities and developmental delays in early childhood. It is unknown whether neonatal morbidities contribute to the increased risk of developmental delay. The objective of this study was to determine the effect of neonatal morbidities after moderately preterm birth on development at preschool age. In a community-based, stratified cohort, parents of 832 moderately preterm children born in 2002 or 2003 completed the Ages and Stage Questionnaire when their child was 43 to 49 months old. Data on Apgar scores, asphyxia, tertiary NICU admission, hospital transfer, circulatory insufficiency, hypoglycemia, septicemia, mechanical ventilation, continuous positive airway pressure, apneas, caffeine treatment, and hyperbilirubinemia were obtained from medical records. We assessed associations of neonatal characteristics with developmental delay, adjusted for gender, small-for-gestational-age status, gestational age, and maternal education. Hypoglycemia and asphyxia were associated with developmental delay; odds ratios (ORs) were 2.42 (95% confidence interval [CI]: 1.23-4.77) and 3.18 (95% CI: 1.01-10.0), respectively. Tertiary NICU admission and hyperbilirubinemia had positive but statistically borderline nonsignificant associations with developmental delay: ORs were 1.74 (95% CI: 0.96-3.15) and 1.52 (95% CI: 0.94-2.46), respectively. No other neonatal morbidities were associated with developmental delay. In multivariate analyses, only hypoglycemia was associated with developmental delay (OR: 2.19; 95% CI: 1.08-4.46). In moderately preterm-born children, only hypoglycemia increased the risk of developmental delay at preschool age. A concerted effort to prevent hypoglycemia might enhance developmental outcome in this group.


Heterogenous graft rejection pathways in class I major histocompatibility complex-disparate combinations and their differential susceptibility to immunomodulation induced by intravenous presensitization with relevant alloantigens.
The present study investigates the heterogeneity of graft rejection pathways in class I major histocompatibility complex (MHC)-disparate combinations and the susceptibility of each pathway to immunomodulation induced by intravenous presensitization with alloantigens. Depletion of CD8+ T cells was induced by repeated administration of anti-CD8 monoclonal antibody. CD8+ T cell-depleted mice failed to generate anti-allo class I MHC cytotoxic T cell (CTL) responses but exhibited anti-allo class I MHC T cell responses, such as mixed lymphocyte reaction (MLR)/IL-2 production, that were induced by CD4+ T cells. In contrast, donor-specific intravenous presensitization (DSP), as a model of donor-specific transfusion, induced almost complete elimination of CD4+ and CD8+ T cell-mediated MLR/IL-2 production, whereas this regimen did not affect the generation of CTL responses induced by DSP-resistant elements (CD8+ CTL precursors and CD4+ CTL helpers). Prolongation of skin graft survival was not induced by either of the above two regimens alone, but by the combination of these. Prolonged graft survival was obtained irrespective of whether the administration of anti-CD8 antibody capable of eliminating CTL was started before or after DSP. The combination of DSP with injection of anti-CD4 antibody also effectively prolonged graft survival. However, this was the case only when the injection of antibody was started before DSP, because such antibody administration was capable of inhibiting the generation of CTL responses by eliminating DSP-resistant CD4+ CTL helpers. These results indicate that (a) the graft rejection in class I-disparate combinations is induced by CD8+ CTL-involved and -independent pathways that are resistant and susceptible to DSP, respectively; (b) DSP contributes to, but is not sufficient for, the prolongation of graft survival; and (c) the suppression of graft rejection requires an additional treatment for reducing DSP-resistant CTL responses. The results are discussed in the context of potential clinical application in attempts to inhibit the generation of DSP-resistant CTL responses upon the prospective DSP.


[Several biochemical signs of loss and restoration of visual function].
With the aid of cytophotometry the author studied the cellular chemistry of neurons (according to the activity of glutamate dehydrogenase) in the III, IV and V layers of the visual cortex in rats during the period of rehabilitation of specific functions following long-term light deprivation. It was demonstrated that a 2-week stay of deprivated animals in conditions of usual light is accompanied by visible normalization of the glutamate dehydrogenase activity in the cortical neurons. However, a complete rehabilitation of the control level was not attained. The study demonstrated differences in the reactions of identical neurons in the IV and V layers to the rehabilitation of specific functions. It is being assumed that the rehabilitation of the visual function may be due to biochemical reconstructions in the monomodel and certain parts of the polymodal neurons in the visual cortex.


Penetrating wounds to the torso: evaluation with triple-contrast multidetector CT.
Penetrating injuries account for a large percentage of visits to emergency departments and trauma centers worldwide. Emergency laparotomy is the accepted standard of care in patients with a penetrating torso injury who are not hemodynamically stable and have a clinical indication for exploratory laparotomy, such as evisceration or gastrointestinal bleeding. Continuous advances in technology have made computed tomography (CT) an indispensable tool in the evaluation of many patients who are hemodynamically stable, have no clinical indication for exploratory laparotomy, and are candidates for conservative treatment. Multidetector CT may depict the trajectory of a penetrating injury and help determine what type of intervention is necessary on the basis of findings such as active arterial extravasation and major vascular, hollow viscus, or diaphragmatic injuries. Because multidetector CT plays an increasing role in the evaluation of patients with penetrating wounds to the torso, the radiologists who interpret these studies should be familiar with the CT findings that mandate intervention.


Reproductive costs of heat shock protein in transgenic Drosophila melanogaster.
Senescence may evolve when genes have antagonistic effects between early reproduction and later age-specific mortality. Although widely consistent with data of quantitative genetics, this model has yet to be validated with the identification of a specific locus presenting such trade-offs. The molecular chaperone hsp70 may be a candidate for such a gene. Heat induced expression of the Hsp70 protein in adults decreases rates of age-specific mortality during normal aging, while maternally experienced heat shock depresses the production of mature progeny. Here we show that maternal heat shock reduces the proportion of egg hatch but not the viability of successfully hatched offspring. To assess whether heat induced maternal expression of hsp70 causes reduced egg hatch, we measured the proportion of eggs that hatch from females engineered to overexpress hsp70 transgenes. We used the same transgenic strains that extend longevity upon hsp70 expression and found that Hsp70 is sufficient to suppress egg hatch. The proportion of egg hatch as a function of hsp70 expression was not reduced in the first eggs laid after maternal heat shock, but appears in later laid eggs, which were at preoogenic and early vitellogenic stages during the maternal expression of hsp70. The contervailing effects of hsp70 upon fecundity and subsequent age-specific mortality exemplify antagonistic pleiotropy, and this trade-off could contribute to the evolution of Drosophila senescence.


[Cost structure of a telephone-based case management in primary care depression therapy].
Case management by health care assistants in small primary care practices has proven effective in improving depression symptoms in depressive patients. Resource consumption and costs of this intervention have not been evaluated yet. Health care assistants and general practitioners of 15 participating practices were interviewed for intervention-related time expenditures. Intervention costs were based on individual labour costs. The annual time spent was 6.3 h per patient for health care assistants and 3.6 h for general practitioners (mean 8.9 patients per practice). The case management caused costs of € 277 per patient and year. A telephone-based case management as add-on therapy improves quality of primary care depression therapy while causing only moderate costs for health care providers.


Factors associated with hospital length of stay and hospital charges of motor vehicle crash related hospitalizations among children in the United States.
To calculate national estimates of motor vehicle crash (MVC)-related hospitalization and associated use of health care resources among patients 20 years and younger and to explore the effects of certain sociodemographic and health care system-related factors and injury severity on use of hospital resources and lengths of stay (LOSs) in the United States. Data from the 2003 Healthcare Cost and Utilization Project Kids' Inpatient Database were used. Pediatric inpatient care in 3438 hospitals in 36 US states. Patients Patients 20 years and younger hospitalized with MVC-related injuries. National estimates of MVC-associated hospitalizations, rates, resource use, Injury Severity Scores, and demographics were calculated. Potentially significant covariate associations were studied using hospital charges and LOS. Sixty-two thousand eight hundred eighty MVC-related hospitalizations occurred among patients 20 years and younger in the United States in 2003, resulting in more than $2.0 billion (SD = $1.2 million) in inpatient charges and 304 196 days (SD = 55,113 days) of hospitalization. Mean (SD) hospital charges and LOS were $33,440 ($55,113) and 4.8 (7.7) days, respectively. The mean (SD) Injury Severity Score was 10.3 (11.4). Adolescents aged 18 through 20 years had the highest hospitalization rates (197 cases per 100,000 children). Older age, being male, urban hospital location, mortality during hospitalization, higher injury severity, and longer LOS were significantly associated with higher total charges. Longer LOS was significantly associated with older age, urban hospital location, higher injury severity, and mortality. Motor vehicle crash-related injuries among children burden health care resources, with nationwide charges exceeding $2 billion annually.


Upregulation of rat lung Na-K-ATPase during hyperoxic injury.
A major function of the alveolar epithelium is to keep the airspace free of fluid and preserve gas exchange. Since Na-K-ATPase is believed to be important in this process, we hypothesized that Na-K-ATPase in the rat lung would increase in response to acute lung injury with pulmonary edema. Na-K-ATPase localization, mRNA expression, and protein levels were determined in hyperoxic lung injury. Adult male rats were exposed to greater than 97% oxygen for 60 h followed by recovery in room air. At 60 h of hyperoxia, the wet-to-dry lung weights increased, consistent with edema. Within the alveolar capillary region, the sodium pump remained localized to the type II cell basolateral membrane by immunocytochemistry. By Northern blot analysis, the level of total lung mRNA expression of the alpha 1- and beta-subunits of Na-K-ATPase increased three- to fourfold during hyperoxia compared with unexposed rats. Total lung Na-K-ATPase membrane protein, visualized with a Western blot technique, appeared to increase by 24 h of hyperoxic insult when compared with levels in unexposed animals. The increase in sodium pump gene expression that occurs during hyperoxic insult, followed by an increase in sodium pump membrane protein, suggests that type II cells increase their Na-K-ATPase synthesis as an early response to pulmonary edema and/or hyperoxia.


Endoscopic Appearance and Management of Recto-Sigmoid Endometriosis: Case Report.
Endometriosis is the presence of endometrial glands outside the uterine cavity. Endometriosis can involve any part of the bowel but most commonly affects the recto-sigmoid part of the bowel. This case shows the endoscopic views of bowel endometriosis and also elaborates on how to approach and manage similar cases of bowel endometriosis. Patients with bowel endometriosis are better managed and treated with a multidisciplinary team including gastroenterologist, gynecologist and colorectal surgeon.


Small vessel disease and cognitive impairment: The relevance of central network connections.
Central brain network connections greatly contribute to overall network efficiency. Here we examined whether small vessel disease (SVD) related white matter alterations in central brain network connections have a greater impact on executive functioning than alterations in non-central brain network connections. Brain networks were reconstructed from diffusion-weighted MRI scans in 72 individuals (75 ± 8 years) with cognitive impairment and SVD on MRI. The centrality of white matter connections in the network was defined using graph theory. The association between the fractional anisotropy (FA) of central versus non-central connections, executive functioning, and markers of SVD was evaluated with linear regression and mediation analysis. Lower FA in central network connections was more strongly associated with impairment in executive functioning than FA in non-central network connections (r = 0.41 vs. r = 0.27; P < 0.05). Results were consistent across varying thresholds to define the central subnetwork (>50%-10% connections). Higher SVD burden was associated with lower FA in central as well as non-central network connections. However, only central network FA mediated the relationship between white matter hyperintensity volume and executive functioning [change in regression coefficient after mediation (95% CI): -0.15 (-0.35 to -0.02)]. The mediation effect was not observed for FA alterations in non-central network connections [-0.03 (-0.19 to 0.04)]. These findings suggest that the centrality of network connections, and thus their contribution to global network efficiency, appears to be relevant for understanding the relationship between SVD and cognitive impairment. Hum Brain Mapp 37:2446-2454, 2016. © 2016 Wiley Periodicals, Inc.


Backscatter laser depolarization studies of simulated stratospheric aerosols: crystallized sulfuric acid droplets.
The optical depolarizing properties of simulated stratospheric aerosols were studied in laboratory laser (0.633 microm) backscattering experiments for application to polarization lidar observations. Clouds composed of sulfuric acid solution droplets, some treated with ammonia gas, were observed during evaporation. The results indicate that the formation of minute ammonium sulfate particles from the evaporation of acid droplets produces linear depolarization ratios of delta approximately 0.02, but delta approximately 0.10-0.15 are generated from acid droplet crystallization effects associated with recycled aerosols and the introduction of ammonia gas into the chamber. It is concluded that partially crystallized sulfuric acid droplets are a likely candidate for explaining the lidar delta approximately 0.10 values that have been observed in the lower stratosphere in the absence of the relatively strong backscattering from homogeneous sulfuric acid droplet (delta approximately 0) or ice crystal (delta approximately 0.5) clouds.


Unpaid Professional Work at Home and Work-Life Interference among Employees with Care Responsibilities.
Employees with caregiving responsibilities often experience work-life interference (WLI), particularly when caring for either disabled persons and/or children. This study examines sample of 288 working Australians from the AWALI national survey data, who care for at least one family member or friend with long-term physical or mental illness, disability, or aging-related problems. We investigated the role of unpaid work at home in predicting WLI, based on a model that included indirect association via inferred causes for working unpaid hours at home and a conditional direct relationship based on number of children. The findings supported our prediction that unpaid work at home is positively associated with WLI but its effect is moderated by number of children. There was a conditional direct effect where employees with care responsibilities experienced a stronger relationship between unpaid hours and WLI when having more children. Further, when the perceived reason for unpaid work was excessively demanding work, the relationship with WLI was stronger. Implications for workers with multiple caregiving responsibilities are discussed.


Plasma ghrelin levels during exercise in healthy subjects and in growth hormone-deficient patients.
To characterise plasma levels of the recently identified endogenous ligand for the GH secretagogue receptor (ghrelin) during submaximal aerobic exercise in healthy adults and in GH-deficient adults. Eight healthy males (mean+/-s.e. age, 40.8+/-2.9 years) and eight hypopituitary males with verified GH deficiency (mean+/-s.e. age, 40.8+/-4.7 years) underwent a baseline test of their peak aerobic capacity (VO(2) peak) and lactate threshold (LT) on a cycle ergometer, as well as an evaluation of body composition. The patients were then studied on two occasions in random order when they exercised for 45 min at their LT. On one occasion, GH replacement had been discontinued from the evening before, whereas on the other occasion they received their evening GH in addition to an intravenous infusion of GH (0.4 IU) during exercise the following day. The healthy subjects exercised at their LT on one occasion without GH. The patients were significantly more obese and had lower VO(2) max (corrected for body weight) and LT as compared with the control subjects. Exercise induced a peak in serum GH concentrations after 45 min in the control group (11.43+/-3.61 microg/l). Infusion of GH in the patients resulted in a peak level after 45 min, whereas no increase was detected when exercising without GH (9.77+/-2.40 (GH) vs 0.11+/-0.07 microg/l (no GH)). Plasma ghrelin levels did not change significantly with time in either study, and no correlations were detected between ghrelin levels and parameters such as GH and IGF-I levels, age or body composition. Plasma ghrelin levels were significantly lower during the study period with GH as compared with the study with no GH. Submaximal aerobic exercise of an intensity sufficient to stimulate GH release was not associated with significant alterations in plasma ghrelin concentrations, which indicated that systemic ghrelin is not involved in the exercise-induced stimulation of GH secretion. The observation that ghrelin levels were lower during GH replacement suggests that GH may feedback-inhibit systemic ghrelin release.


Phage-typing patterns and lysogenicity of methicillin-resistant strains of Staphylococcus aureus from Sydney, Australia, 1965-85.
Methicillin-resistant strains of Staphylococcus aureus isolated at the Royal Prince Alfred Hospital since 1965 were differentiated by phage-typing and by their lysogenic status. Most of these strains were isolated during two periods, 1965-72 and 1976-85. Nearly all of the strains isolated in the first period had one of four phage-typing patterns. Strains with each typing pattern carried two prophages; these eight phages were all different, as characterised by serological grouping and lytic spectrum. Lysogenisation of the non-lysogenic strain 1489 with each of these phages narrowed its phage-typing pattern; the typing pattern of the double lysogens was generally similar to and occasionally identical with that of the host strain that had yielded the pair of phages. In the second period, strains with one of five other phage-typing patterns predominated. Representatives of each of these carried the lysogenic phage C. The first methicillin-resistant strain carrying this phage had been isolated in 1974. The current methicillin-resistant S. aureus strains thus appear to form a distinct group that can be differentiated from those seen in earlier years.


[Investigation of Hepatitis C Virus (HCV) Genotypes by "Reverse Hybridisation Strip Assay" and DNA Sequence Analysis Methods].
Hepatitis C Virus (HCV) is a virus that is estimated to infect approximately 185 million people worldwide and causes 350 000 deaths each year. The target in HCV treatment is the sustained virological response (SVR), and the most important virological factors determining SVR are genotype and baseline HCV-RNA levels. In this study, it was aimed to compare the HCV genotypes and subtypes detected by the "line probe assay (LIPA)" method with sequence analysis. HCV genotypes and subtypes were investigated by line probe assay (LIPA) (NLM analytica, Italy) and sequence analysis methods in 212 patients with chronic HCV diagnosis and with HCV RNA viral load of 10⁴ IU/ml and above. 5'UTR and core regions were studied in LIPA method and NS5B gene region was studied in the sequencing method. After amplifying 340 bp region in the NS5B gene region with the hemi-nested PCR method, sequence analysis (ABI 3500 Genetic Analyzer [Applied Biosystems, USA]) was performed. Statistical analysis of the study was determined by using TURCOSA Analytics program. In the study, HCV genotypes and subtypes that were determined in 212 patients by LIPA method were; 40 (18.8%) genotype 1a, 97 (45.75%) genotype 1b, 7 (3.3%) genotype 2, 12 (5.6%) genotype 2a/c, 2 (0.94%) genotype 2b, 19 (8.96%) genotype 3, 16 (7.55%) genotype 4, 1 (0.47%) genotype 4a, 15 (7.5%) genotype 4c/d, 1 (0.47%) genotype 4h and 2 (0.94%) genotype 5. The HCV genotypes and subtypes determined in 212 patients by sequence analysis were; 20 (9.43%) genotype 1a, 118 (55.6%) genotype 1b, 16 (7.55%) genotype 2a, 4 (1.89%) genotype 2b, 1 (0.47%) genotype 2k, 21 (9.91%) genotype 3a, 2 (0.94%) genotype 4a, 28 (13.21%) genotype 4d and 2 (0.94%) genotype 5a. The difference in results between the two methods was found to be statistically significant (p< 0.001). According to the viral load-genotype relationship, the highest viral load was detected in genotype 1a patients (p< 0.001). In conclusion, in order to determine HCV genotypes and subtypes in our study, LIPA and sequence analysis methods were compared and the genotype compatibility between the two methods were determined as 97.5% on the basis of genotype and 19% on the basis of subtype. Since the direct-acting antiviral (DEA) treatment protocol in chronic HCV patients is planned according to the genotype/subtype determination, the inconsistency of the results obtained in the LIPA method, which is routinely widely used, with the sequence analysis method is remarkable, and it was concluded that these results should be supported by studies containing more samples.


Synthesis of an insulin-loaded mucoadhesive nanoparticle designed for intranasal administration: focus on new diffusion media.
Intranasal administration is a drug delivery approach to provide a non-invasive pharmacological response in the central nervous system with relatively small peripheral side effects. To improve the residence time of intranasal drug delivery systems in the nasal mucosa, mucoadhesive polymers (e.g., chitosan) can be used. Here, insulin-loaded chitosan nanoparticles were synthesized and their physiochemical properties were evaluated based on requirements of intranasal administration. The nanoparticles were spherical (a hydrodynamic diameter of 165.3 nm, polydispersity index of 0.24, and zeta potential of +21.6 mV) that granted mucoadhesion without any noticeable toxicity to the nasal tissue. We applied a new approach using the Krebs-Henseleit buffer solution along with simulated nasal fluid in a Franz's diffusion cell to study this intranasal drug delivery system. We used the Krebs-Henseleit buffer because of its ability to supply glucose to the cells which serves as a novel ex vivo diffusion medium to maintain the viability of the tissue during the experiment. Based on diffusion rate and histopathological endpoints, the Krebs-Henseleit buffer solution can be a substituent solution to the commonly used simulated nasal fluid for such drug delivery systems.


[Lipid vehicles for the parenteral administration of drugs (1): emulsions].
To increase the efficiency of medication used in therapy and diagnostics, there have been, during the last decades, important advances in the inclusion techniques hereof, in vehicles which give greater selectivity and tolerance. Among the colloidal vehicle systems of medications with different medications, the object of the present review study. Therefore, we analyze the rationale of their use as a vehicle of medication, their physical and chemical characteristics, the ways of elimination after their administration, as well as the formulations in oil-water emulsions for the parenteral administration of diferent active substances such as diazepam, propofol, methohexital, ethomidate, amphotericin B, cytostatics, and cyclosporine, among others. The results of published studies on the efficiency and therapeutic applications of these are presented.


Enzyme-substrate intermediate formation at lysine 329 of human deoxyhypusine synthase.
Deoxyhypusine (Nepsilon-(4-aminobutyl)lysine) is the key intermediate in the posttranslational synthesis of the unique amino acid, hypusine (Nepsilon-(4-amino-2-hydroxybutyl)lysine). Deoxyhypusine synthase catalyzes the formation of deoxyhypusine by conjugation of the butylamine moiety of spermidine to the epsilon-amino group of one specific lysine residue of the eukaryotic translation initiation factor 5A (eIF-5A) precursor protein. However, in the absence of the eIF-5A precursor, catalysis involves only the NAD-dependent cleavage of spermidine to generate 1,3-diaminopropane and a putative 4-carbon amine intermediate that gives rise to Delta1-pyrroline. We have obtained evidence for a covalent enzyme-substrate intermediate that accumulates in the absence of the eIF-5A precursor. Incubation of human recombinant enzyme with [1, 8-3H]spermidine and NAD, followed by reduction with NaBH3CN, resulted in specific radiolabeling of the enzyme. The radioactive component in the reduced enzyme intermediate was identified as deoxyhypusine and was shown to occur at a single locus. The fact that labeled deoxyhypusine was found after treatment with a reducing agent suggests an intermediate with the butylamine moiety derived from spermidine attached through an imine linkage to the epsilon-amino group of a specific lysine residue of the enzyme. This residue has been identified as lysine 329. Separate experiments showing efficient transfer of labeled butylamine moiety from enzyme intermediate to eIF-5A precursor strongly support a reaction mechanism involving an imine intermediate.


Axon guidance in the mouse optic chiasm: retinal neurite inhibition by ephrin "A"-expressing hypothalamic cells in vitro.
In the mammalian visual system, retinal axons undergo temporal and spatial rearrangements as they project bilaterally to targets on the brain. Retinal axons cross the neuraxis to form the optic chiasm on the hypothalamus in a position defined by overlapping domains of regulatory gene expression. However, the downstream molecules that direct these processes remain largely unknown. Here we use a novel in vitro paradigm to study possible roles of the Eph family of receptor tyrosine kinases in chiasm formation. In vivo, Eph receptors and their ligands distribute in complex patterns in the retina and hypothalamus. In vitro, retinal axons are inhibited by reaggregates of isolated hypothalamic, but not dorsal diencephalic or cerebellar cells. Furthermore, temporal retinal neurites are more inhibited than nasal neurites by hypothalamic cells. Addition of soluble EphA5-Fc to block Eph "A" subclass interactions decreases both the inhibition and the differential response of retinal neurites by hypothalamic reaggregates. These data show that isolated hypothalamic cells elicit specific, position-dependent inhibitory responses from retinal neurites in culture. Moreover, these responses are mediated, in part, by Eph interactions. Together with the in vivo distributions, these data suggest possible roles for Eph family members in directing retinal axon growth and/or reorganization during optic chiasm formation.


Multiple imputation of incomplete multilevel data using Heckman selection models.
Missing data is a common problem in medical research, and is commonly addressed using multiple imputation. Although traditional imputation methods allow for valid statistical inference when data are missing at random (MAR), their implementation is problematic when the presence of missingness depends on unobserved variables, that is, the data are missing not at random (MNAR). Unfortunately, this MNAR situation is rather common, in observational studies, registries and other sources of real-world data. While several imputation methods have been proposed for addressing individual studies when data are MNAR, their application and validity in large datasets with multilevel structure remains unclear. We therefore explored the consequence of MNAR data in hierarchical data in-depth, and proposed a novel multilevel imputation method for common missing patterns in clustered datasets. This method is based on the principles of Heckman selection models and adopts a two-stage meta-analysis approach to impute binary and continuous variables that may be outcomes or predictors and that are systematically or sporadically missing. After evaluating the proposed imputation model in simulated scenarios, we illustrate it use in a cross-sectional community survey to estimate the prevalence of malaria parasitemia in children aged 2-10 years in five regions in Uganda.


[Epilepsy surgery in children with focal cortical dysplasias].
Focal cortical dysplasias (FCD) are the first cause of surgery in paediatric epilepsy surgery. The pre-surgical assessment in FCD is often complex, since they are lesions that can be highly epileptogenic and at the same time can preserve neurological functioning and may not be displayed in magnetic resonance imaging. The success of the operation largely depends on the proper identification of the lesion and the possibility of performing a complete resection of the dysplastic tissue. In this work we review the literature related with this topic, in relation to the authors' experience. The study reviews some of the advances made as regards the pre-surgical assessment and the neurosurgical management of epilepsy in children with FCD; results from the surgical series regarding the classifications of FCD and the post-surgical prognostic factors are commented on; some anatomo-clinical phenotypes that are distinctive in children with FCD and their surgical management are described; and current challenges and the future of the surgical treatment of epilepsy in FCD are also briefly discussed. The advances being made in the methods of pre-surgical diagnosis and surgical procedures are making it possible to offer successful treatment at earlier ages in patients with FCD who were previously considered 'non-lesional' and in patients with localised lesions in the 'eloquent cortex'. The identification of anatomo-electro-clinical phenotypes of FCD makes it possible to establish surgical approaches and post-surgical prognostic expectations that are well suited to each situation, which are better in the transmantle-type FCD IIb and in bottom-of-sulcus dysplasias than in multilobe FCD, which are mostly FCD I.


Improving pharmaceutical innovation by building a more comprehensive database on drug development and use.
New drugs and biologics have had a tremendous impact on the treatment of many diseases. However, available measures suggest that pharmaceutical innovation has remained relatively flat, despite substantial growth in research and development spending. We review recent literature on pharmaceutical innovation to identify limitations in measuring and assessing innovation, and we describe the framework and collaborative approach we are using to develop more comprehensive, publicly available metrics for innovation. Our research teams at the Brookings Institution and Deerfield Institute are collaborating with experts from multiple areas of drug development and regulatory review to identify and collect comprehensive data elements related to key development and regulatory characteristics for each new molecular entity approved over the past several decades in the United States and the European Union. Subsequent phases of our effort will add data on downstream product use and patient outcomes and will also include drugs that have failed or been abandoned in development. Such a database will enable researchers to better analyze the drivers of drug innovation, trends in the output of new medicines, and the effect of policy efforts designed to improve innovation.


Voltage-Gated Sodium Channels Regulating Action Potential Generation in Itch-, Nociceptive-, and Low-Threshold Mechanosensitive Cutaneous C-Fibers.
We evaluated the effect of voltage-gated sodium channel 1 (NaV1) blockers in three nonoverlapping C-fiber subtypes in the mouse skin: chloroquine (CQ)-sensitive C-fibers with high mechanical thresholds-itch C-fibers; second, CQ-insensitive, capsaicin-sensitive C-fibers with high mechanical thresholds-nociceptors; and CQ and capsaicin-insensitive C-fibers with a very low mechanical threshold-C-LTMs. NaV1-blocking drugs were applied to the nerve terminal receptive fields using an innervated isolated dorsal mouse skin-nerve preparation where the drugs are delivered into the skin intra-arterially. We combined these studies with an analysis of the mRNA expression of the α-subunits of NaV1 in individual dorsal root ganglia neurons labeled from the same region of the skin. Our results show that virtually all nociceptors and itch C-fibers expressed the tetrodotoxin (TTX)-resistant channels NaV1.8 and NaV1.9. However, TTX applied selectively into the skin abolished the action potential firing in response to mechanical stimulation in 75% of the itch C-fibers, 100% of the nociceptors, and 100% of C-LTMs. NaV1.7 was the most commonly expressed TTX-sensitive NaV1 in all three C-fiber subtypes innervating the dorsal skin. Selectively blocking NaV1.7 abolished responses in about 40% of itch C-fibers, 65% of nociceptors, but only 20% of C-LTMs. Blocking NaV1.8 alone had no affect on the firing sensitivity of the C-fibers. However, in itch and nociceptive C-fibers where the activation was not inhibited with a NaV1.7 blocker, adding the NaV1.8 blocker silenced action potential discharge.


Responses to 19 generations of litter size selection in the Nebraska Index line. I. Reproductive responses estimated in pure line and crossbred litters.
Our objective was to estimate responses in reproductive traits in the Nebraska Index line (I) after 19 generations of selection for increased litter size. Responses were estimated in dams producing pure line, F1, and three-way cross litters. A total of 850 litters were produced over six year-seasons, including 224 pure line litters, 393 F1 litters produced from I and C females mated with Danbred NA Landrace (L) or Duroc-Hampshire (T) boars, and 233 litters by F1 L x I and L x C females mated with T boars. Contrasts of means were used to estimate the genetic difference between I and C and interactions of line differences with mating type. Farrowing rates of lines I (u = 91.0%) and C (u = 92.8%) did not differ. Averaged across all genetic groups, mean number born alive per litter was 10.1 pigs, and number and weight of pigs weaned per litter, both adjusted for number nursed and weaning age of 12 d, were 9.7 pigs and 34.4 kg, respectively. Averaged across mating types, direct genetic effects of I were greater than C (P < 0.05) for total born (3.53 pigs), number born alive (2.53 pigs), number of mummified pigs (0.22 pig), and litter birth weight (2.14 kg). The direct genetic effect of line I was less than C (P < 0.05) for litter weaning weight (-1.88 kg). Interactions of line effects with crossing system were significant (P < 0.05) for total number born, number of stillborn pigs, number weaned, and litter weaning weight. In pure line litters, I exceeded C by 4.18 total pigs and 1.76 stillborn pigs per litter, whereas the estimate of I-C in F1 litters was 2.74 total pigs and 0.78 stillborn pig per litter. The contrast between I and C for number weaned and litter weaning weight in pure litters was 0.32 pig and -0.28 kg, respectively, compared with 0.25 pig and -2.14 kg in F1 litters. Crossbreeding is an effective way to use the enhanced reproductive efficiency of the Index line.


Spectrally encoded fiber-based structured lighting probe for intraoperative 3D imaging.
Three dimensional quantification of organ shape and structure during minimally invasive surgery (MIS) could enhance precision by allowing the registration of multi-modal or pre-operative image data (US/MRI/CT) with the live optical image. Structured illumination is one technique to obtain 3D information through the projection of a known pattern onto the tissue, although currently these systems tend to be used only for macroscopic imaging or open procedures rather than in endoscopy. To account for occlusions, where a projected feature may be hidden from view and/or confused with a neighboring point, a flexible multispectral structured illumination probe has been developed that labels each projected point with a specific wavelength using a supercontinuum laser. When imaged by a standard endoscope camera they can then be segmented using their RGB values, and their 3D coordinates calculated after camera calibration. The probe itself is sufficiently small (1.7 mm diameter) to allow it to be used in the biopsy channel of commonly used medical endoscopes. Surgical robots could therefore also employ this technology to solve navigation and visualization problems in MIS, and help to develop advanced surgical procedures such as natural orifice translumenal endoscopic surgery.


The complete amino acid sequence of chicken skeletal-muscle enolase.
The complete amino acid sequence of chicken skeletal-muscle enolase, comprising 433 residues, was determined. The sequence was deduced by automated sequencing of hydroxylamine-cleavage, CNBr-cleavage, o-iodosobenzoic acid-cleavage, clostripain-digest and staphylococcal-proteinase-digest fragments. The presence of several acid-labile peptide bonds and the tenacious aggregation of most CNBr-cleavage fragments meant that a commonly used sequencing strategy involving initial CNBr cleavage was unproductive. Cleavage at the single Asn-Gly peptide bond with hydroxylamine proved to be particularly useful. Comparison of the sequence of chicken enolase with the two yeast enolase isoenzyme sequences shows that the enzyme is strongly conserved, with 60% of the residues identical. The histidine and arginine residues implicated as being important for the activity of yeast enolase are conserved in the chicken enzyme. Secondary-structure predictions are analysed in an accompanying paper [Sawyer, Fothergill-Gilmore & Russell (1986) Biochem. J. 236, 127-130].


Parallel gas spectroscopy using mid-infrared supercontinuum from a single Si3N4 waveguide.
Efficient third-order nonlinear optical processes have been successfully integrated on silicon nitride (Si3N4) waveguides. In particular, owing to Si3N4 wide transparency window spanning from the visible to the middle-infrared (mid-IR), efficient mid-IR dispersive-wave (DW) generation from a fiber laser has been recently demonstrated, and its potential as a source for absorption spectroscopy of a single gas has been established. Here we show that the system can be further engineered to broaden the coverage of a single DW without losing efficiency, as to enable simultaneous and discrete detection of several gas-phase molecules within the 2900 and 3380cm-1 functional group region. We demonstrate quantitative detection of acetylene, methane, and ethane using a simple direct-absorption spectroscopy scheme, achieving a several hundreds of parts-per-million noise-equivalent detection limit with a 5 cm long gas cell.


Biochemical transfer of relational responding (transposition).
Hooded rats were trained to choose the larger of two circles in order to obtain a liquid reward. Recipients of brain extract from these trained donors showed a significant preference for an even larger circle when given a nonreinforced test for several days after the injection of extract. The substrate for relational learning may be biochemically transferred.


Mental Health and Quality of Life for Healthcare Workers in a University Hospital Under COVID-19.
The aim of this study was to assess the psychosocial characteristics of the employees working at a university hospital and investigated the factors affecting their quality of life (QOL) under COVID-19. This study enrolled 1,191 healthcare workers from a university hospital, including doctors, nurses, administrative officer and technicians. Besides demographic information, depression, anxiety, somatization, insomnia, resilience, and QOL were assessed. The nurses presented significantly higher scores for anxiety, depression and showed significantly higher insomnia scores and significantly lower resilience scores. The occupations showed significant differences in the QOL and sub-groups, including the overall quality of life and general health (F=4.774, p<0.001), psychological domain (F=6.230, p<0.001), and environment domain (F=5.254, p<0.001). There was a positive correlation between the QOL and resilience (r=0.608, p<0.01). However, depression (r=-0.502, p<0.01), anxiety (r=-0.425, p<0.01), somatization (r=-0.364, p<0.01), and insomnia (r=-0.385, p<0.01) showed negative correlations with the QOL. Resilience was the most important factor influencing the QOL. The results of this study showed that low resilience adversely affected the QOL and the mental health of the healthcare workers, which consequently had a direct effect on the quality of medical care given to patients.


[Evaluation of fetal protective-adaptive reactions in gestosis and pelvic presentation].
Biophysical profile of the fetus (BPF) was determined with the use of ultrasonic scanning and cardiotocography in 21 pregnant women with the pelvic presentation of the fetus and 32 patients with gestosis. Uteroplacental and fetoplacental circulation were also under study. The results of studies were analyzed together with the data on the labor outcome and course of the neonatal period. The findings evidence a regular depression of some BPF elements in hypoxia. Circulatory disorders in gestosis are initiated by disorders of the uteroplacental circulation, followed by involvement of the fetoplacental circulation in this process. Reduced BPF parameters in pelvic presentation of the fetus evidence functional immaturity of the fetus. A close relationship between the results of BPF assessment and fetal Doppler ultrasonics data has been revealed.


Connecting EEG signal decomposition and response selection processes using the theory of event coding framework.
The neurophysiological mechanisms underlying the integration of perception and action are an important topic in cognitive neuroscience. Yet, connections between neurophysiology and cognitive theoretical frameworks have rarely been established. The theory of event coding (TEC) details how perceptions and actions are associated (bound) in a common representational domain (the "event file"), but the neurophysiological mechanisms underlying these processes are hardly understood. We used complementary neurophysiological methods to examine the neurophysiology of event file processing (i.e., event-related potentials [ERPs], temporal EEG signal decomposition, EEG source localization, time-frequency decomposition, EEG network analysis). We show that the P3 ERP component and activity modulations in inferior parietal regions (BA40) reflect event file binding processes. The relevance of this parietal region is corroborated by source localization of temporally decomposed EEG data. We also show that temporal EEG signal decomposition reveals a pattern of results suggesting that event file processes can be dissociated from pure stimulus and response-related processes in the EEG signal. Importantly, it is also documented that event file binding processes are reflected by modulations in the network architecture of theta frequency band activity. That is, when stimulus-response bindings in event files hamper response selection this was associated with a less efficient theta network organization. A more efficient organization was evident when stimulus-response binding in event files facilitated response selection. Small-world network measures seem to reflect event file processing. The results show how cognitive-theoretical assumptions of TEC can directly be mapped to the neurophysiology of response selection.


The co-crystal N,N'-bis-[(pyridin-1-ium-2-yl)meth-yl]ethane-dithio-amide bis-(2,6-di-nitro-benzoate)-2,6-di-nitro-benzoic acid (1/4).
The asymmetric unit of title co-crystal, C14H16N4S2 (2+)·2C7H3N2O6 (-)·4C7H4N2O6, comprises a centrosymmetric dipyridinium dication, a 2,6-di-nitro-benzoate anion and two independent 2,6-di-nitro-benzoic acid mol-ecules. The pyridin-ium rings are each approximately perpendicular to the central di-thio-amide unit [dihedral angle = 80.67 (12)°]. The carboxyl-ate/carb-oxy-lic acid groups are approximately perpendicular to the benzene ring to which they are attached [dihedral angles = 78.85 (16), 81.46 (19) and 71.28 (15)°]. By contrast, the major twist exhibited by a nitro group is manifested in a dihedral angle of 32.66 (17)°. The most prominent feature of the crystal packing is linear supra-molecular chains along [1-10], featuring O-H⋯O(carboxyl-ate) and pyridinium-N-H⋯O hydrogen bonds. These are consolidated into a three-dimensional architecture by thio-amide-nitro N-H⋯O, C-H⋯O and π-π [inter-centroid distance = 3.524 (2) Å] inter-actions. One of the nitro O atoms was refined over two sites; the major site was 0.65 (7) occupied.


Differential effect of ionizing radiation on transcription in repair-deficient and repair-proficient mice.
Experiments were designed to examine in vivo changes in total transcription and in the expression of the c-fos gene following whole-body exposure of mice to JANUS fission-spectrum neutrons. Radiation repair-deficient (wst/wst) and -proficient (wst/., C57BL/6 x C3H F1) mice were exposed to JANUS fission-spectrum neutrons calibrated to deliver a gut dose of 50 cGy. Animals were sacrificed less than 10 or at 60 min postirradiation, and gut tissues were removed for study. Our results revealed that, in repair-proficient mice, an immediate depression (relative to untreated control) in total transcription was evident that continued through 1 h postirradiation. Conversely, radiation-sensitive wst/wst mice displayed doubled transcription levels postirradiation. Expression of c-fos was consistently depressed following radiation exposure in control and wst/wst mice. However, the depression of c-fos mRNA was delayed in wst/wst mice relative to controls. These results demonstrate abnormal regulation of transcription and of c-fos mRNA accumulation in repair-deficient wasted mice following exposure to ionizing radiation. In addition, this work documents rapid total transcriptional depression in normal mice following radiation exposure.


Limbal palisades of Vogt.
The palisades of Vogt are distinctive normal features of the human corneoscleral limbus. Our clinical studies indicate that they are more discrete in younger and in more heavily pigmented individuals, and that they appear more regular and prominent at the lower limbus than at the upper limbus. They are seen only infrequently in the horizontal meridian. There is some symmetry (though it is not exact) from one eye to the other in the same person. The anatomy of the palisades appears to be unique for a given individual. In this respect, as well as in their microscopic anatomy, the palisades of Vogt appear comparable to fingerprints, and the term "conjunctivoglyphics" ("conjunctival carvings") or "limboglyphics" is suggested in analogy with "dermatoglyphics." The palisades of Vogt have a distinct vasculature with narrow, barely visible, arterial and venous components of radially oriented hairpin loops. Angiography reveals that these vessels leak fluorescein relatively late and only to a moderate extent. They respond to inflammation by dilatation and gross breakdown of their physiologic barrier properties. The functions of the palisades of Vogt are not known with certainty, but their interpalisadal epithelial rete ridges may serve as a repository for corneal epithelial cells. They may thus be important in both aging and diseases of the cornea.