Hyperthermic Intraperitoneal Chemotherapy-Induced Molecular Changes in Humans Validate Preclinical Data in Ovarian Cancer.
Hyperthermic intraperitoneal chemotherapy (HIPEC) confers a survival benefit in epithelial ovarian cancer (EOC) and in preclinical models. However, the molecular changes induced by HIPEC have not been corroborated in humans. A feasibility trial evaluated clinical and safety outcomes of HIPEC with cisplatin during optimal cytoreductive surgery (CRS) in patients with EOC diagnosed with stage III, IV, or recurrent EOC. Pre- and post-HIPEC biopsies were comprehensively profiled with genomic and transcriptomic sequencing to identify mutational and RNAseq signatures correlating with response; the tumor microenvironment was profiled to identify potential immune biomarkers; and transcriptional signatures of tumors and normal samples before and after HIPEC were compared to investigate HIPEC-induced acute transcriptional changes. Thirty-five patients had HIPEC at the time of optimal CRS; all patients had optimal CRS. The median progression-free survival (PFS) was 24.7 months for primary patients and 22.4 for recurrent patients. There were no grade 4 or 5 adverse events. Anemia was the most common grade 3 adverse event (43%). Hierarchical cluster analyses identified distinct transcriptomic signatures of good versus poor responders to HIPEC correlating with a PFS of 29.9 versus 7.3 months, respectively. Among good responders, significant HIPEC-induced molecular changes included immune pathway upregulation and DNA repair pathway downregulation. Within cancer islands, % programmed cell death protein 1 expression in CD8+ T cells significantly increased after HIPEC. An exceptional responder (PFS 58 months) demonstrated the highest programmed cell death protein 1 increase. Heat shock proteins comprised the top differentially upregulated genes in HIPEC-treated tumors. Distinct transcriptomic signatures identify responders to HIPEC, and preclinical model findings are confirmed for the first time in a human cohort.


No evidence for the use of DIR, D-D fusions, chromosome 15 open reading frames or VH replacement in the peripheral repertoire was found on application of an improved algorithm, JointML, to 6329 human immunoglobulin H rearrangements.
Antibody diversity is created by imprecise joining of the variability (V), diversity (D) and joining (J) gene segments of the heavy and light chain loci. Analysis of rearrangements is complicated by somatic hypermutations and uncertainty concerning the sources of gene segments and the precise way in which they recombine. It has been suggested that D genes with irregular recombination signal sequences (DIR) and chromosome 15 open reading frames (OR15) can replace conventional D genes, that two D genes or inverted D genes may be used and that the repertoire can be further diversified by heavy chain V gene (VH) replacement. Safe conclusions require large, well-defined sequence samples and algorithms minimizing stochastic assignment of segments. Two computer programs were developed for analysis of heavy chain joints. JointHMM is a profile hidden Markow model, while JointML is a maximum-likelihood-based method taking the lengths of the joint and the mutational status of the VH gene into account. The programs were applied to a set of 6329 clonally unrelated rearrangements. A conventional D gene was found in 80% of unmutated sequences and 64% of mutated sequences, while D-gene assignment was kept below 5% in artificial (randomly permutated) rearrangements. No evidence for the use of DIR, OR15, multiple D genes or VH replacements was found, while inverted D genes were used in less than 1 per thousand of the sequences. JointML was shown to have a higher predictive performance for D-gene assignment in mutated and unmutated sequences than four other publicly available programs. An online version 1.0 of JointML is available at http://www.cbs.dtu.dk/services/VDJsolver.


A Gja1 missense mutation in a mouse model of oculodentodigital dysplasia.
Oculodentodigital dysplasia (ODDD) is an autosomal dominant disorder characterized by pleiotropic developmental anomalies of the limbs, teeth, face and eyes that was shown recently to be caused by mutations in the gap junction protein alpha 1 gene (GJA1), encoding connexin 43 (Cx43). In the course of performing an N-ethyl-N-nitrosourea mutagenesis screen, we identified a dominant mouse mutation that exhibits many classic symptoms of ODDD, including syndactyly, enamel hypoplasia, craniofacial anomalies and cardiac dysfunction. Positional cloning revealed that these mice carry a point mutation in Gja1 leading to the substitution of a highly conserved amino acid (G60S) in Cx43. In vivo and in vitro studies revealed that the mutant Cx43 protein acts in a dominant-negative fashion to disrupt gap junction assembly and function. In addition to the classic features of ODDD, these mutant mice also showed decreased bone mass and mechanical strength, as well as altered hematopoietic stem cell and progenitor populations. Thus, these mice represent an experimental model with which to explore the clinical manifestations of ODDD and to evaluate potential intervention strategies.


Representation of three-dimensional visual space in the cerebral cortex.
This article reviews two issues relevant to the topic of how three-dimensional space is represented in the cerebral cortex. The first is the question of how individual neurons encode information that might contribute to stereoscopic estimation of visual depth. Particular attention is given to the current understanding of the neural representation of motion through three-dimensional space and to the complexities that arise in interpreting neuronal responses to this complex stimulus parameter. The second issue considered is the disorderlines that exists in the retinotopic mapping of the visual field in some cortical visual areas. Several extrastriate areas have been found to contain maps of the contralateral visual hemifield that are disorderly in the sense that the representation of various parts of the visual field are often misplaced or grossly over-or under-represented. It is suggested that this disorderlines may in some cases represent adaptations to facilitate certain types of visual functions.


Increasing sharp safety device use in healthcare: a semi-structured interview study.
The use of sharp safety devices in healthcare is considered the most important means of preventing occupational percutaneous injuries and has been mandated for use in most hospitals in industrialized countries including in Canada. However, clinical personnel's perceptions on the use of safety devices needs further characterization to improve compliance. This study's objective was to identify healthcare provider perspectives on different aspects of sharp safety device use and on how use could be increased. Using a constant comparison approach, data from semi-structured interviews with 39 nurses, physicians and phlebotomists providing direct patient care, and six nurses acting as the interface between clinical personnel and purchasing departments, were analyzed. Study participants were from three of the six health authorities in British Columbia. The four major categories that emerged from the data were: selection processes; identification and replacement; training; and multi-level barriers and facilitators. Findings highlighted the importance of including personnel regularly using safety devices at each stage of their selection including when they are being considered for replacement with superior devices, as well as the need for appropriate initial and refresher training, and how practices at the hospital, ward and individual level facilitate safety device use.


Adipokines and Inflammation Alter the Interaction Between Rheumatoid Arthritis Synovial Fibroblasts and Endothelial Cells.
Objective: The long-distance migration of rheumatoid arthritis synovial fibroblasts (RASFs) in the severe combined immunodeficiency (SCID) mouse model of rheumatoid arthritis (RA) suggests that an interaction between RASFs and endothelial cells (EC) is critical in this process. Our objective was to assess whether immunomodulatory factors such as adipokines and antirheumatic drugs affect the adhesion of RASFs to ECs or the expression of surface molecules. Methods: Primary ECs or human umbilical vein endothelial cell (HUVEC) and primary RASFs were stimulated with adiponectin (10 μg/mL), visfatin (100 ng/mL), and resistin (20 ng/mL) or treated with methotrexate (1.5 and 1,000 μM) and the glucocorticoids prednisolone (1 μM) and dexamethasone (1 μM), respectively. The expression of adhesion molecules was analyzed by real-time polymerase chain reaction. The interaction of both cell types was analyzed under static (cell-to-cell binding assay) and dynamic conditions (flow-adhesion assay). Results: Under static conditions, adipokines increased mostly binding of RASFs to EC (adiponectin: 40%, visfatin: 28%, tumor necrosis factor α: 49%). Under flow conditions, visfatin increased RASF adhesion to HUVEC (e.g., 0.5 dyn/cm2: 75.2%). Reduced adhesion of RASFs to E-selectin was observed after treatment with dexamethasone (e.g., 0.9 dyn/cm2: -40%). In ECs, tumor necrosis factor α (TNF-α) increased expression of intercellular adhesion molecule 1 (20-fold) and vascular cell adhesion molecule 1 (77-fold), whereas P-selectin was downregulated after stimulation with TNF-α (-6-fold). Conclusion: The adhesion of RASFs to EC was increased by visfatin under static and flow conditions, whereas glucocorticoids were able to decrease adhesion to E-selectin. The process of migration and adhesion of RASFs to ECs could be enhanced by adipokines via adhesion molecules and seems to be targeted by therapeutic intervention with glucocorticoids.


CONBERCEPT IN PATIENTS WITH TREATMENT-NAIVE NEOVASCULAR AGE-RELATED MACULAR DEGENERATION IN REAL-LIFE SETTING IN CHINA.
In this study, we aimed to evaluate the efficacy and safety of intravitreal conbercept in patients with treatment-naive neovascular age-related macular degeneration in real-life setting. Three consecutive intravitreal injections of conbercept following a pro re nata protocol. The main outcomes were the changes of Early Treatment Diabetic Retinopathy Study best-corrected visual acuity and central retinal thickness between the baseline and the 12th month. Mean best-corrected visual acuity was improved from 39.39 ± 24.91 letters at the baseline to 44.26 ± 22.89 letters at the final follow-up (P < 0.001). At the 12th month, the proportion of optimal response was 43.48% compared with 36.96% of poor response and 19.56% of nonresponse. A mean central retinal thickness of 480.94 ± 178.47 μm at the baseline was significantly reduced to 366.33 ± 173.52 μm at the 12th month. Patients received a median of 5.32 intravitreal injections. At the 12th month, the mean change in best-corrected visual acuity of eyes with intraretinal cystoid fluid from the baseline was markedly lower than that of eyes without intraretinal cystoid fluid. No adverse events were attributed to conbercept. With 12-month follow-up, conbercept was proved to be an effective and safety treatment for patients with treatment-naive neovascular age-related macular degeneration in real-life setting.


Instructional technology adoption of medical faculty in teaching.
Despite large investment by higher education institutions in technology for faculty and student use, instructional technology is not being integrated into instructions in the higher education institutions, including medical education institutions. While diffusion of instructional technologies has been reached a saturation point with early adopters of technology, it has remained limited among mainstream faculty. This study explored technology adoption patterns and perceptions of medical faculty about barriers and incentives to technology adoption in teaching.Complete data was obtained from 155 participants by using survey methodology and analyzed on the basis of theories of diffusion of innovation.Findings provided evidence for limited adoption of relatively new tools associated with instruction into mainstream faculty. Inadequate hardware for students and faculty, lack of reward structure, insufficient traing oppurtunites were identified as major barriers to faculty technology adoption.


Comparison of the clinical effectiveness of oseltamivir and zanamivir against influenza virus infection in children.
We compared the clinical effectiveness of oseltamivir and zanamivir in children with influenza A (H1N1) virus, influenza A (H3N2) virus, and influenza B virus infections. Total febrile period and the duration of fever after the start of treatment were compared between an oseltamivir-treated group (mean age, 8.9 years; range, 4.0-15.9 years) and a zanamivir-treated group (mean age, 10.0 years; range, 4.0-15.7 years) in the pediatric outpatient clinics of our hospitals. Oseltamivir was used to treat 91 children with influenza A (H3N2) infection and 24 children with influenza A (H1N1) infection. Zanamivir was used to treat 35 children with influenza A (H3N2) infection and 12 children with influenza A (H1N1) infection. Oseltamivir was also used to treat 128 children with influenza B virus infection, and zanamivir was used to treat 59 with influenza B virus infection. There was no statistically significant difference in total febrile period or duration of fever after the start of treatment between the oseltamivir-treated group and the zanamivir-treated group of children with influenza A (H3N2) infection (mean duration of febrile period, 2.40 days vs. 2.39 days; mean duration of fever after the start of treatment, 1.35 days vs. 1.40 days), influenza A (H1N1) (mean duration of febrile period, 2.60 days vs. 2.46 days; mean duration of fever after the start of treatment, 1.79 days vs, 1.54 days), or influenza B (mean duration of febrile period, 2.95 days vs. 2.84 days; mean duration of fever after the start of treatment, 1.86 days vs. 1.67 days). Oseltamivir was more effective against influenza A (H3N2) than against influenza A (H1N1) or influenza B. Oseltamivir and zanamivir were equally effective in reducing the febrile period of children with influenza A (H1N1), influenza A (H3N2), and influenza B virus infection.


Shared and distinct neural correlates of first and second language morphological processing in bilingual brain.
While morphology constitutes a crucial component of the human language system, the neural bases of morphological processing in the human brain remains to be elucidated. The current study aims at exploring the extent to which the second language (L2) morphological processing would resemble or differ from that of their first language (L1) in adult Chinese-English bilinguals. Bilingual participants were asked to complete a morphological priming lexical decision task drawing on derivational morphology, which is present for both Chinese and English, when their electrophysiological and optical responses were recorded concurrently. Functional near-infrared spectroscopy (fNIRS) revealed a neural dissociation between morphological and semantic priming effects in the left fronto-temporal network, while L1 Chinese engaged enhanced activation in the left prefrontal cortex for morphological parsing relative to L2 English. In the early stage of lexical processing, cross-language morphological processing manifested a difference in degree, not in kind, as revealed by the early left anterior negativity (ELAN) effect. In addition, L1 and L2 shared both early and late structural parsing processes (P250 and 300 ~ 500 ms negativity, respectively). Therefore, the current results support a unified competition model for bilingual development, where bilinguals would primarily employ L1 neural resources for L2 morphological representation and processing.


Thermal sensitivity of bacteriocytes constrains the persistence of intracellular bacteria in whitefly symbiosis under heat stress.
Temperature affects the persistence of diverse symbionts of insects. Our previous study indicates that the whitefly symbionts confined within bacteriocytes or scattered throughout the body cavity outside bacteriocytes may have differential thermal sensitivity. However, the underlying mechanisms remain largely unknown. Here, we report that following continuous heat stress, Portiera and Hamiltonella were almost completely depleted in two species of Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) of the Bemisia tabaci whitefly cryptic species complex. Meanwhile, proliferation of bacteriocytes was severely inhibited and approximately 50% of the nymphs had lost one of the two bacteriomes. While cell size of bacteriocytes was increased, cell number was severely decreased leading to reduction of total volume of bacteriocytes. Moreover, bacteriocyte organelles and associated symbionts were lysed, and huge amount of electron-dense inclusions accumulated. Eventually, Portiera and Hamiltonella failed to be transmitted to the next generation. In contrast, Rickettsia could be detected although at a reduced level, and successfully transmitted to eggs. The results suggest that the thermal sensitivity of bacteriocytes may limit thermal tolerance and vertical transmission of the associated symbionts, and consequently different patterns of distribution of symbionts may affect their capacity to tolerate unfavourable temperatures and persistence in the host.


A Role for the RNA Polymerase Gene Specificity Factor σ54 in the Uniform Colony Growth of Uropathogenic Escherichia coli.
The canonical function of a bacterial sigma (σ) factor is to determine the gene specificity of the RNA polymerase (RNAP). In several diverse bacterial species, the σ54 factor uniquely confers distinct functional and regulatory properties on the RNAP. A hallmark feature of the σ54-RNAP is the obligatory requirement for an activator ATPase to allow transcription initiation. Different activator ATPases couple diverse environmental cues to the σ54-RNAP to mediate adaptive changes in gene expression. Hence, the genes that rely upon σ54 for their transcription have a wide range of different functions suggesting that the repertoire of functions performed by genes, directly or indirectly affected by σ54, is not yet exhaustive. By comparing the growth patterns of prototypical enteropathogenic, uropathogenic, and nonpathogenic Escherichia coli strains devoid of σ54, we uncovered that the absence of σ54 results in two differently sized colonies that appear at different times specifically in the uropathogenic E. coli (UPEC) strain. Notably, UPEC bacteria devoid of individual activator ATPases of the σ54-RNAP do not phenocopy the σ54 mutant strain. Thus, it seems that σ54's role as a determinant of uniform colony appearance in UPEC bacteria represents a putative non-canonical function of σ54 in regulating genetic information flow. IMPORTANCE RNA synthesis is the first step of gene expression. The multisubunit RNA polymerase (RNAP) is the central enzyme responsible for RNA synthesis in bacteria. The dissociable sigma (σ) factor subunit directs the RNAP to different sets of genes to allow their expression in response to various cellular needs. Of the seven σ factors in Escherichia coli and related bacteria, σ54 exists in a class of its own. This study has uncovered that σ54 is a determinant of the uniform growth of uropathogenic E. coli on solid media. This finding suggests a role for this σ54 in gene regulation that extends beyond its known function as an RNAP gene specificity factor.


Relationship between Gs alpha messenger ribonucleic acid splice variants and the molecular forms of Gs alpha protein in rat brown adipose tissue.
Recent work indicates that multiple molecular forms of Gs alpha can be produced by alternative splicing of a primary Gs alpha transcript. This study examined the pattern of expression of messenger RNAs (mRNAs) encoding the large (Gs alpha-L) and small (Gs alpha-S) molecular forms of Gs alpha in several rat tissues, including brown adipose tissue. In all tissues examined, Gs alpha-L mRNA was most abundant. This was most pronounced for brain tissue (95% of total), and least for brown adipose tissue (57%). The pattern of Gs alpha isoform expression of a given tissue was strongly correlated with the Gs alpha mRNA splicing pattern in that tissue. Cold exposure and surgical denervation are treatments that have been shown to, respectively, increase or decrease total Gs alpha mRNA in rat brown adipose tissue. Despite producing up to 4-fold differences in total Gs alpha mRNA, these treatments did not affect the proportions of Gs alpha mRNA splice variants in brown fat. In contrast, perinatal stimulation of brown fat was associated with changes in the splicing pattern of Gs alpha mRNA, and these changes were reflected in Gs alpha protein expression. Specifically, during the perinatal period Gs alpha-L mRNA increased significantly but Gs alpha-L protein levels did not change, whereas Gs alpha-S mRNA did not change and Gs alpha-S protein levels significantly declined. These data provide further evidence that the increased Gs alpha mRNA levels that occur in interscapular brown adipose tissue during periods of stimulation serve to maintain membrane levels of Gs alpha protein.


Regulation of tau phosphorylation and protection against beta-amyloid-induced neurodegeneration by lithium. Possible implications for Alzheimer's disease.
Alzheimer's disease is a neurodegenerative disorder characterized by the accumulation of the beta-amyloid peptide and the hyperphosphorylation of the tau protein, among other features. The most widely accepted hypothesis on the etiopathogenesis of this disease proposes that the aggregates of the beta-amyloid peptide are the main triggers of tau hyperphosphorylation and the subsequent degeneration of affected neurons. In support of this view, fibrillar aggregates of synthetic beta-amyloid peptide induce tau hyperphosphorylation and cell death in cultured neurons. We have previously reported that lithium inhibits tau hyperphosphorylation and also significantly protects cultured neurons from cell death triggered by beta-amyloid peptide. As lithium is a relatively specific inhibitor of glycogen synthase kinase-3 (in comparison with other protein kinases), and other studies also point to a relevant role of this enzyme, we favor the view that glycogen synthase kinase-3 is a crucial element in the pathogenesis of Alzheimer's disease. In our opinion, the possibility of using lithium, or other inhibitors of glycogen synthase kinase-3, in experimental trials aimed to ameliorate neurodegeneration in Alzheimer's disease should be considered.


Management of avascular necrosis of the femoral head--an overview.
The most common cause of AVN of the femoral head is a displaced subcapital or transcervical fracture. Circumstances here differ significantly from those encountered in the patient with nontraumatic AVN, especially because the traumatic variety is seen most often in the elderly population. Attempts to preserve the femoral head are generally not indicated and the treatment of choice for the hip with sufficient pain and disability is either endoprosthetic replacement or total hip replacement. In contradistinction, nontraumatic AVN occurs primarily in younger adults and is often bilateral. In these patients the goal is to preserve, not to replace, the femoral head. "Conservative" or nonsurgical management has generally been unsuccessful. Results with established surgical procedures have been inconsistent and frequently disappointing, often because they have been instituted too late. Although no approach is completely effective, in general the earlier treatment begins, the better the results will be. Early diagnosis, therefore, is essential. This depends on a heightened clinical awareness of this condition. The importance of various imaging techniques, including high-quality plain radiographs, technetium scans, and magnetic resonance imaging must be stressed. Newer procedures for the treatment of early AVN are being developed and evaluated. These currently include the use of vascularized grafts, osteochondral allografts, transtrochanteric rotational osteotomy, and electrical stimulation. I hope that one or more of these approaches will prove useful in preserving the femoral head. Until such time, I shall continue to use bone grafting with decompression to treat hips with limited involvement. When definite collapse of the femoral head has occurred and when pain and disability are sufficient to require surgical intervention, the treatment of choice is arthroplasty. Although many still favor the use of a bipolar femoral endoprosthesis, I believe that total hip replacement provides more consistent, more durable, and better results. Whichever arthroplasty is chosen, I anticipate continually improving results with advances in technique and component design.


Toward the influence of temporal attention on the selection of targets in a visual search task: An ERP study.
We used ERPs to investigate whether temporal attention interacts with spatial attention and feature-based attention to enhance visual processing. We presented a visual search display containing one singleton stimulus among a set of homogenous distractors. Participants were asked to respond only to target singletons of a particular color and shape that were presented in an attended spatial position. We manipulated temporal attention by presenting a warning signal before each search display and varying the foreperiod (FP) between the warning signal and the search display in a blocked manner. We observed distinctive ERP effects of both spatial and temporal attention. The amplitudes for the N2pc, SPCN, and P3 were enhanced by spatial attention indicating a processing benefit of relevant stimulus features at the attended side. Temporal attention accelerated stimulus processing; this was indexed by an earlier onset of the N2pc component and a reduction in reaction times to targets. Most importantly, temporal attention did not interact with spatial attention or stimulus features to influence visual processing. Taken together, the results suggest that temporal attention fosters visual perceptual processing in a visual search task independently from spatial attention and feature-based attention; this provides support for the nonspecific enhancement hypothesis of temporal attention.


Limited role of the c-Jun N-terminal kinase pathway in a neonatal rat model of cerebral hypoxia-ischemia.
D-JNKI1, a cell-permeable peptide inhibitor of the c-Jun N-terminal kinase (JNK) pathway, has been shown to be a powerful neuroprotective agent after focal cerebral ischemia in adult mice and young rats. We have investigated the potential neuroprotective effect of D-JNKI1 and the involvement of the JNK pathway in a neonatal rat model of cerebral hypoxia-ischemia (HI). Seven-day-old rats underwent a permanent ligation of the right common carotid artery followed by 2 h of hypoxia (8% oxygen). Treatment with D-JNKI1 (0.3 mg/kg intraperitoneally) significantly reduced early calpain activation, late caspase 3 activation and, in the thalamus, autophagosome formation, indicating an involvement of JNK in different types of cell death: necrotic, apoptotic, and autophagic. However, the size of the lesion was unchanged. Further analysis showed that neonatal HI induced an immediate decrease in JNK phosphorylation (reflecting mainly JNK1 phosphorylation) followed by a slow progressive increase (including JNK3 phosphorylation 54 kDa), whereas c-jun and c-fos expression were both strongly activated immediately after HI. In conclusion, unlike in adult ischemic models, JNK is only moderately activated after severe cerebral HI in neonatal rats and the observed positive effects of D-JNKI1 are insufficient to give neuroprotection. Thus, for perinatal asphyxia, D-JNKI1 can only be considered in association with other therapies.


Yeast-expressed recombinant As16 protects mice against Ascaris suum infection through induction of a Th2-skewed immune response.
Ascariasis remains the most common helminth infection in humans. As an alternative or complementary approach to global deworming, a pan-anthelminthic vaccine is under development targeting Ascaris, hookworm, and Trichuris infections. As16 and As14 have previously been described as two genetically related proteins from Ascaris suum that induced protective immunity in mice when formulated with cholera toxin B subunit (CTB) as an adjuvant, but the exact protective mechanism was not well understood. As16 and As14 were highly expressed as soluble recombinant proteins (rAs16 and rAs14) in Pichia pastoris. The yeast-expressed rAs16 was highly recognized by immune sera from mice infected with A. suum eggs and elicited 99.6% protection against A. suum re-infection. Mice immunized with rAs16 formulated with ISA720 displayed significant larva reduction (36.7%) and stunted larval development against A. suum eggs challenge. The protective immunity was associated with a predominant Th2-type response characterized by high titers of serological IgG1 (IgG1/IgG2a > 2000) and high levels of IL-4 and IL-5 produced by restimulated splenocytes. A similar level of protection was observed in mice immunized with rAs16 formulated with alum (Alhydrogel), known to induce mainly a Th2-type immune response, whereas mice immunized with rAs16 formulated with MPLA or AddaVax, both known to induce a Th1-type biased response, were not significantly protected against A. suum infection. The rAs14 protein was not recognized by A. suum infected mouse sera and mice immunized with rAs14 formulated with ISA720 did not show significant protection against challenge infection, possibly due to the protein's inaccessibility to the host immune system or a Th1-type response was induced which would counter a protective Th2-type response. Yeast-expressed rAs16 formulated with ISA720 or alum induced significant protection in mice against A. suum egg challenge that associates with a Th2-skewed immune response, suggesting that rAS16 could be a feasible vaccine candidate against ascariasis.


Dairy calf mortality rate: influence of management and housing factors on calf mortality rate in Tulare County, California.
Data on calf mortality, calving site, calf-rearing facilities, and calf-management procedures were collected from 16 dairy farms in Tulare County, California. Discriminant analyses were used to test if any significant differences in these factors existed between farms with higher than average and farms with lower than average death losses. The average mortality rate over a period of at least 2 years on individual farms varied from a low of 3.5 +/- 1.1% to a high of 30.6 +/- 3.1%. Calf management personnel was the only factor significantly related to the mortality rate, with considerably fewer death losses on farms where the owner managed the calves than on farms where employees performed these duties. In general, other factor categories relating to size of the cow herd, calving site, and calf housing were not related significantly to calf deaths. The average number of heifer calves born each year on individual farms was increasing year by year, and on some farms, there seemed to be a concomitant increase in death losses. This, together with the tendency toward greater number of deaths on the larger owner-managed farms, indicated that calf care was diluted under these conditions.


Culture revisited: A reply to comments.
Replies to comments by Winston & Maher (see record 2015-45553-005), Abi-Hashem (see record 2015-45553-006), and Hook & Watkins (see record 2015-45553-007), on the original article by Christopher et al. (see record 2014-20055-001). In this brief response, the authors clarify some elements of their thinking and address some misconceptions put forward by the commentators.


Expansion of wastewater-based disease surveillance to improve health equity in California's Central Valley: sequential shifts in case-to-wastewater and hospitalization-to-wastewater ratios.
Over a third of the communities (39%) in the Central Valley of California, a richly diverse and important agricultural region, are classified as disadvantaged-with inadequate access to healthcare, lower socio-economic status, and higher exposure to air and water pollution. The majority of racial and ethnic minorities are also at higher risk of COVID-19 infection, hospitalization, and death according to the Centers for Disease Control and Prevention. Healthy Central Valley Together established a wastewater-based disease surveillance (WDS) program that aims to achieve greater health equity in the region through partnership with Central Valley communities and the Sewer Coronavirus Alert Network. WDS offers a cost-effective strategy to monitor trends in SARS-CoV-2 community infection rates. In this study, we evaluated correlations between public health and wastewater data (represented as SARS-CoV-2 target gene copies normalized by pepper mild mottle virus target gene copies) collected for three Central Valley communities over two periods of COVID-19 infection waves between October 2021 and September 2022. Public health data included clinical case counts at county and sewershed scales as well as COVID-19 hospitalization and intensive care unit admissions. Lag-adjusted hospitalization:wastewater ratios were also evaluated as a retrospective metric of disease severity and corollary to hospitalization:case ratios. Consistent with other studies, strong correlations were found between wastewater and public health data. However, a significant reduction in case:wastewater ratios was observed for all three communities from the first to the second wave of infections, decreasing from an average of 4.7 ± 1.4 over the first infection wave to 0.8 ± 0.4 over the second. The decline in case:wastewater ratios was likely due to reduced clinical testing availability and test seeking behavior, highlighting how WDS can fill data gaps associated with under-reporting of cases. Overall, the hospitalization:wastewater ratios remained more stable through the two waves of infections, averaging 0.5 ± 0.3 and 0.3 ± 0.4 over the first and second waves, respectively.


Clinical documentation and occurrence of putative periodontopathic bacteria in human immunodeficiency virus-associated periodontal disease.
Human immunodeficiency virus (HIV)-associated gingivitis (HIV-G) and HIV-associated periodontitis (HIV-P) are two intraoral lesions manifested by patients with HIV infection. Periodontal indices were measured for 87 subjects in 5 study groups: HIV-seropositive patients with healthy periodontium (HIV-H), with HIV-G, or with HIV-P; and non-HIV-infected subjects with healthy periodontium (H) or with adult chronic periodontitis (P). The quantitative clinical parameters were compared and statistically significant intergroup differences were noted. The mean scores on PI and PD do not discriminate between HIV-seropositive and non-HIV-infected seronegative cohorts, but a significant difference in the GI between HIV-H and H was noted. When categories of PD and AL are examined, some differences become apparent. Generally, the PD and AL of HIV-P are not as great as those of P. PI correlates well with GI (r = 0.86) in P, but does not (r = 0.33) in HIV-P. In addition, the occurrence of selected putative periodontopathic bacteria (Porphyromonas gingivalis, spirochetes, and motile eubacteria) in these lesions was determined by brightfield (after staining), darkfield and immunofluorescent microscopy. No difference in microbiological profile in the bacterial groups monitored was found between P and HIV-P. Spirochetes were found to be more abundant than P. gingivalis in the lesions of P and HIV-P. In marked contrast, P. gingivalis was found to be in highest numbers in samples from the gingival crevice of H as determined by indirect immunofluorescence.


[Appendiceal mucocele disclosed by a psoas tumor. Apropos of a case].
Mucocele of the appendix is a rare lesion. Rupture of appendicular mucocele into the psoas is uncommon. We report a case of a 77-year-old man, with psoas tumor on abdominopelvic ultrasonography and computed tomography. The preoperative diagnosis was type III hydatid cyst of the psoas, psoas tumor or retroperitoneal mass. Surgical exploration revealed an appendicular mucocele ruptured into the psoas. Appendicectomy was performed with an uneventful postoperative course after 3 years. The authors discuss the diagnostic and therapeutic problems raised by this rare lesion.


Bacterial phosphotransferase system (PTS) in carbohydrate uptake and control of carbon metabolism.
More than 20 carbohydrates may be transported into the bacterial cell by the phosphoenopyruvate:carbohydrate phosphotransferase system (PTS) that is widely spread among bacteria. The PTS consists of two cytoplasmic energy-coupling proteins (Enzyme I and HPr) and a range of carbohydrate-specific Enzymes II, which catalyze concomitant carbohydrate translocation and phosphorylation. The phosphorylation status of PTS components reflects the availability of carbohydrates and the energy conditions of the cell. In many bacteria, PTS and the associated proteins convert this information to signals, which transduced through different mechanisms lead to phenomena of catabolite repression, inducer control or chemotaxis. These features of PTS provide bacteria with an integrated system, which assures optimal utilization of carbohydrates in complex environments. Furthermore, some bacteria evolved parallel systems that serve a regulatory functions, but apparently do not catalyze the carbohydrate transport. Here we review the findings that recently advanced the understanding of various aspects of PTS-dependent carbohydrate transport and regulation of bacterial catabolism.


Central retinal vein obstruction in a neonate occurring during laser photocoagulation treatment for retinopathy of prematurity.
A premature infant who developed central retinal vein obstruction in one eye during laser therapy for high-risk prethreshold retinopathy of prematurity is described. Systemic investigations were done to detect the cause of such an acute and unreported episode. Acquired deficiency of protein C levels was detected. Premature infants are at risk of thromboembolic episodes due to deranged hematological parameters. Acquired protein C deficiency possibly precipitated acute retinal vein occlusion during laser therapy in this infant. Intravitreal bevacizumab was used successfully to treat the episode of retinal venous occlusion. Retinal vein obstruction during laser treatment for retinopathy of prematurity is reported for the first time.


Which physicians and practices are using electronic medical records?
Greater use of electronic medical records (EMRs) has the potential to improve the quality of medical care and has become a goal of the U.S. administration. According to the National Ambulatory Medical Care Survey, only an average of 17.6 percent of physicians reported use of EMRs in their office-based practices. The adoption rate did not increase from 2001 through 2003. Practices with more physicians and those owned by health maintenance organizations (HMOs) were significantly more likely to use this technology, but use varied little by the characteristics of individual physicians, the practice's scope of services, or the practice's sources of revenue.


RNA turnover and chromatin-dependent gene silencing.
Over the last few years, there has been a convergence of two seemingly disparate fields of study: chromatin-dependent gene silencing and RNA turnover. In contrast to RNA turnover mechanisms that operate on a truly posttranscriptional level, we are at the beginning of studies leading the way toward a model in which RNA turnover mechanisms are also involved in chromatin-dependent gene regulation. In particular, data from a variety of organisms have shown that the assembly of silent chromatin coincides with the presence or absence of non-protein-coding RNAs (ncRNAs). These range from long ncRNAs that have been classically implicated in the regulation of dosage compensation and genomic imprinting to small ncRNAs which are involved in heterochromatin assembly via the RNA interference (RNAi) pathway. This raises the question of how common ncRNAs are used to control gene expression at the level of chromatin. It is known at least, that they are present, as recent findings indicate that transcription of eukaryotic genomes is much more widespread than previously anticipated. However, the existence of a ncRNA does not prove its biological significance. Thus, a future challenge will be to distinguish the ncRNAs that are in some way meaningful to the organism from those that arise from the imperfect fidelity of the transcription machinery. Finally, no matter whether functional or not, RNAs transcribed from supposedly silent chromatin seem to be processed rapidly. Recent data from both fission and budding yeast suggest that chromatin-dependent gene silencing is achieved, at least in part, through RNA turnover mechanisms that use components of the RNAi pathway as well as polyadenylation-dependent RNA decay. Hence, silent chromatin is not only controlled transcriptionally, but also on co- and posttranscriptional levels.


Comparison of signature lipid methods to determine microbial community structure in compost.
The microbial community structure changes substantially during the composting process and simple methods to follow these changes can potentially be used to estimate compost maturity. In this study, two such methods, the microbial identification (MIDI) method and the ester-linked (EL) procedure to determine the composition of long-chain fatty acids, were applied to compost samples of different age. The ability of the two methods to describe the microbial succession was evaluated by comparison with phospholipid fatty acid (PLFA) analysis on the same samples.Samples were taken from a 200-l laboratory compost reactor, treating source-separated organic household waste. During the initial stages of the process, the total concentration of fatty acids in compost samples treated with the EL and MIDI methods was many times higher than with the PLFA method. This was probably due to the presence of fatty acids from the organic material in the original waste. However, this substantial difference between PLFA and the other two methods was not found later in composting. Although the PLFA method gave the most detailed information about the growth and overall succession of the microbial community, the much simpler MIDI and EL methods also successfully described the shift from the initially dominating straight chain fatty acids to iso- and anteiso branched, 10 Me branched and cyclopropane fatty acids in the later stages of the process. Thus, the MIDI and EL extraction methods appear to be suitable for analysis of microbial FAME profiles in compost, particularly in the later stages of the process.


[Alterations in blood circulation of vesicourethral segment in men with neurogenic and non-neurogenic lower urinary tract symptoms].
To specify the pathogenetic link between blood supply of the bladder neck and lower urinary tract symptoms (LUTS). The study involved 78 men aged 26 to 50 years, including 19 patients with multiple sclerosis (MS) and LUTS, 29 patients with chronic prostatitis category IIIB with LUTS and 30 patients with chronic prostatitis category IIIB without LUTS (control group). All the patients underwent Doppler ultrasonography of prostatic arteries and selective study of blood flow in the neck of the bladder. Pharmacological test using combinations of 1-blockers (1-AB) with m-anticholinergics (m-CB) and phosphodiesterase type 5 inhibitors (PDEI-5). The changes in the blood circulation of VUS were found to correlate with LUTS. The response of blood flow depended not only on the type of pharmacological agents, but also on the degree of vascular changes and neurological deficit in VUS. The findings of VUS vascular pharmacological tests with PDEI-5 and 1-AB + m-CB were comparable in chronic prostatitis with non-neurogenic LUTS and in MS with neurogenic LUTS. Vascular reaction in VUS depends on the presence of neurological deficit. The combination of (1-AB+ m-CB) increases the blood circulation of the bladder neck and prostate and reduces the LUTS. PDEI-5 reduces LUTS due to the positive effect on the blood circulation of the prostate and VUS. Circulatory abnormalities (=hypoxia) in VUS is an important pathogenetic mechanism of neurogenic and non-neurogenic LUTS and the way to compensate them using a combination of (1-AB+ m-CB) or PDEI-5. Therefore, one of the mechanisms of LUTS is associated with impaired blood flow in the bladder neck and VUS.


HLA-DRB1*15 confers susceptibility to juvenile SLE but is not associated with disease presentation: an Egyptian Study.
The etiology of Systemic lupus erythematosus (SLE) seems to be multifactorial including environmental as well as genetic factors. Major histocompatibility complex (MHC) genes especially HLA-DRB1 and HLA-DQB1 are strongly implicated in susceptibility to SLE. Moreover ethnicity has been found to have a significant role in both disease susceptibility and disease expression. This study was carried out to determine HLA-DRB1 allele association with SLE susceptibility and disease presentation in Egyptian children with juvenile onset SLE. HLA-DRB1 allele typing was done using polymerase chain reaction-sequence-specific oligonucleotide probe for 65 juvenile Egyptian SLE patients and 150 healthy controls. p-values were corrected for the number of the alleles tested (Pc). HLA-DRB1*15 g allele was significantly increased in SLE children versus controls (OR = 4.76; 95% CI = 1.83-12.4; p = 0.001 and Pc = 0.012). No HLA-DRB1 allele was found to be statistically significant associated with musculoskeletal, cutaneous, hematologic, cardiac or neuropsychiatric manifestations in SLE patients (p > 0.05). Moreover no statistically significant association was found between HLA-DRB1 alleles and clinical presentation or histologic classes of lupus nephritis. The current work suggests that HLA-DRB1*15g allele may be a susceptibility allele in Egyptian children with SLE but is not related to clinical presentation of SLE.


[Monoclonal antibodies CD95 in an acute period of ishemic stroke in arterial hypertension].
The data on examination of 83 patients in acute period of cerebral stroke caused by arterial hypertension and 30 healthy controls are presented. In all the patients, content of Fas expressed lymphocytes (CD95), indirectly suggesting apoptosis, has been determined. The level of Fas/Apo-1+ lymphocytes was significantly higher comparing to the control group. Cerebral stroke severity positively correlated with Fas/Apo-1+ lymphocytes. The higher was a level of monoclonal antibodies CD95, the poorer was the course and prognosis both for recovering and for life. This correlation argues for a role of Fas/Apo-1+ lymphocytes in the apoptotic process, and the level of their expression reflects that of apoptosis in damaged brain tissue and serves as a sign of immune inflammation in response to acute cerebral disaster.


Mandibular midline distraction osteogenesis.
In orthodontics, bone structure, its density and dimensions play an essential role by explaining limitations in magnitude, size and extent of tooth movement. Severe anterior crowding is one of the most frequently encountered dental malocclusions. Its therapy is mostly limited by lack of basal and alveolar bone and it often involves tooth extractions. Mandibular midline distraction osteogenesis is a method of natural bone generation and also a treatment option to achieve space regaining in a much-reduced lower jaw with distinctive frontal place deficit and severe anterior crowding, without sacrificing permanent teeth. McCarthy and Guerrero were of the first researchers reporting on this method applied on human lower jaws and they increased clinical interest in this approach. Although this method has been clinically used ever since, many questions concerning effects on bone regeneration speed, bone quality, tooth movement into regenerated area, periodontal health and long-time stability of treatment outcomes have not been sufficiently investigated. This overview should present the current clinical and biological state of knowledge about bone gain and tooth movement through regenerate bone. Furthermore it should encourage interest in further research on this topic.


Bisphosphonate-associated femur fractures have high complication rates with operative fixation.
Bisphosphonate-associated femur fractures have been well described but the preoperative patient factors, treatment modalities, and complications of treatment are unclear. We asked whether a diagnosis of osteoporosis, the characteristic radiographic features of bisphosphonate-related femur fractures, and complication rates differed in patients with operatively treated femoral shaft fractures receiving bisphosphonates and in patients not receiving bisphosphonates. We retrospectively reviewed 43 patients with bisphosphonate-associated femoral shaft fractures (including subtrochanteric) from 2002 to 2008 and 20 patients with similar fractures but not treated with bisphosphonates. Similar implants were used in both groups, but a greater number of adjuvants were used in the bisphosphonate cohort. We recorded preoperative osteoporosis and radiographic findings of the characteristic bisphosphonate femur fracture and early complications. The minimum followup was 5 months (mean, 29 months; range 5-60 months). Preoperatively a greater percentage of patients treated with bisphosphonates had confirmed osteoporosis than those not treated with bisphosphonates (24% versus 5%, respectively), a greater percentage had a proximal fracture location (48% versus 40%, respectively), and their mean cortex to shaft diameter ratio was greater (24% versus 15%, respectively). The bisphosphonate cohort had a higher rate of intraoperative fractures (21% versus 0%) and postoperative plate failures (30% versus 0%). Despite low rates of other risk factors and ample use of biologic adjuvants, patients treated with bisphosphonates having femur fractures have more complications. Level III, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.


Secoisolariciresinol diglycoside, a flaxseed lignan, exerts analgesic effects in a mouse model of type 1 diabetes: Engagement of antioxidant mechanism.
Peripheral painful neuropathy is one of the most common complications in diabetes and necessitates improved treatment. Secoisolariciresinol diglycoside (SDG), a predominant lignan in flaxseed, has been shown in our previous studies to exert antidepressant-like effect. As antidepressant drugs are clinically used to treat chronic neuropathic pain, this work aimed to investigate the potential analgesic efficacy of SDG against diabetic neuropathic pain in a mouse model of type 1 diabetes. We subjected mice to diabetes by a single intraperitoneal (i.p.) injection of streptozotocin (STZ, 200 mg/kg), and Hargreaves test or von Frey test was used to assess thermal hyperalgesia or mechanical allodynia, respectively. Chronic instead of acute SDG treatment (3, 10 or 30 mg/kg, p.o., twice per day for three weeks) ameliorated thermal hyperalgesia and mechanical allodynia in diabetic mice, and these analgesic actions persisted about three days when SDG treatment was terminated. Although chronic treatment of SDG to diabetic mice did not impact on the symptom of hyperglycemia, it greatly attenuated excessive oxidative stress in sciatic nerve and spinal cord tissues, and partially counteracted the condition of weight decrease. Furthermore, the analgesic actions of SDG were abolished by co-treatment with the reactive oxygen species donor tert-butyl hydroperoxide (t-BOOH), but potentiated by the reactive oxygen species scavenger phenyl-N-tert-butylnitrone (PBN). These findings indicate that chronic SDG treatment can correct neuropathic hyperalgesia and allodynia in mice with type 1 diabetes. Mechanistically, the analgesic actions of SDG in diabetic mice may be associated with its antioxidant activity.


[Direct coronary stent implantation without predilatation--a new therapeutic approach with a special balloon catheter design].
Stent implantation serves as the gold standard for proximal lesions of the coronary arteries with a diameter between 2.75-3.5 mm. Our new concept aims at a reduced procedure duration and fluoro-time as well as a decreased ischemic period during stent implantation. A new therapeutic concept of a direct stent implantation without predilatation was tested using a specially developed balloon catheter on which various 14-16 mm long "slotted-tube" stents are mounted between two conical, radiopaque markers. In 105 consecutive patients, who were scheduled for angioplasty, this method of direct stent implantation was performed. Six of the procedures were performed for acute myocardial infarction and 8 in so-called high-risk procedures. The direct stent implantation was successful in 88%. In 6%, predilatation of the lesion site was necessary before stent placement. In the remaining 6%, a stent could not be successfully implanted despite the availability of various other systems. Comparison of the direct stent implantation with conventional stent placement with predilatation revealed that 1) The fluoro-time for direct stent implantation, compared to the conventional method, was 8.4 +/- 4.9 min vs. 13.7 +/- 8.0 min; p < 0.05, respectively. Furthermore, there were less balloons used per lesion for direct stent implantation (1.4 +/- 0.4) compared to the conventional method (1.7 +/- 0.7), but there was not a significant difference. 2) If we compare those patients with successful direct stent implantation with those with the unsuccessful procedures, the latter group had a higher percent of angiographically visible calcification at the site of the lesion (80% vs. 18%; p < 0.01). In addition, these patients had an increased average age (72 +/- 7 vs. 61 +/- 11 yrs; p < 0.01). The success rate of direct stent implantation did not depend on lesion diameter stenosis before PTCA. Stent dislocation was observed in 3.8% of the procedures, and a single case of stent embolism was seen. In conclusion, the direct stent implantation offers the advantages of a shortened fluoro-time, the use of fewer balloons, and has the potential of less ischemic stress compared to the conventional method of stent implantation with predilatation, if old patients with calcified lesions are excluded. This should be proved on a large scale in future studies also considering a learning curve with regard to the new method. Whether this new approach also reduces the restenosis rate, warrants further studies.


Altered patterns of transcription of the septin gene, SEPT9, in ovarian tumorigenesis.
Ovarian carcinoma represents the most lethal gynaecological malignancy. A variety of morphological subtypes are recognised (e.g. serous, mucinous, endometrioid), which may be benign, borderline or malignant. While their relationship is controversial, knowledge of the molecular mechanisms of ovarian tumorigenesis may help resolve this issue and perhaps identify early markers of disease. Perturbed patterns of expression of the SEPT9 gene on chromosome 17q25.3 have been implicated in a variety of tumour types including both breast and ovarian neoplasia. In preliminary studies, we showed that SEPT9 mRNA was upregulated in a bank of ovarian tumours, which included benign, borderline and malignant tumours, and reported increased levels of one splice variant, SEPT9_v4*. We now describe a comprehensive analysis of SEPT9 expression specifically in serous and mucinous ovarian tumours (benign, borderline and malignant), using cDNA microarray, semi- and quantitative RTPCR of microdissected archival tumour material. Our data show consistent and specific overexpression of both SEPT9_v1 and SEPT9_v4* transcripts in the epithelial component of ovarian tumours. These transcripts show highest levels of expression in serous and mucinous borderline tumours. SEPT9_v1 is also upregulated in both serous and mucinous carcinomas. Interestingly, highest levels of expression are observed in serous borderline and low-grade tumours rather than high-grade in keeping with a model of progression of benign, borderline and low-grade serous tumours.


[How clinicians make choices.]
Humans are influenced by irrational preferences in making choices, so that clinicians also are prone to irrational decision-making. Exhaustive information about the value of therapies are frequently unavailable, full results of intervention trials are often hidden, conclusive evidence is highly desirable but it is seldom accessible. Healthcare could benefit from the new ways of approaching the decision framework due to behavioral economists: understanding the hidden traps in decision-making and the way the human brain works is an essential step and the best protection from biases is always awareness.


Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of beta-tubulin.
Allyl sulfides are characteristic flavor components obtained from garlic. These sulfides are thought to be responsible for their epidemiologically proven anticancer effect on garlic eaters. This study was aimed at clarifying the molecular basis of this anticancer effect of garlic by using human colon cancer cell lines HCT-15 and DLD-1. The growth of the cells was significantly suppressed by diallyl trisulfide (DATS, HCT-15 IC50 = 11.5 microM, DLD-1 IC50 = 13.3 microM); however, neither diallyl monosulfide nor diallyl disulfide showed such an effect. The proportion of HCT-15 and that of DLD-1 cells residing at the G1 and S phases were decreased by DATS, and their populations at the G2/M phase were markedly increased for up to 12 h. The cells with a sub-G1 DNA content were increased thereafter. Caspase-3 activity was also dramatically increased by DATS. Fluorescence-activated cell sorter analysis performed on the cells arrested at the G1/S boundary revealed cell cycle-dependent induction of apoptosis through the transition of the G2/M phase to the G1 phase by DATS. DATS inhibited tubulin polymerization in an in vitro cell-free system. DATS disrupted microtubule network formation of the cells, and microtubule fragments could be seen at the interphase. Peptide mass mapping by liquid chromatography-tandem mass spectrometry analysis for DATS-treated tubulin demonstrated that there was a specific oxidative modification of cysteine residues Cys-12beta and Cys-354beta to form S-allylmercaptocysteine with a peptide mass increase of 72.1 Da. The potent antitumor activity of DATS was also demonstrated in nude mice bearing HCT-15 xenografts. This is the first paper describing intracellular target molecules directly modified by garlic components.


Alternative splicing of the Ca2+ channel beta4 subunit confers specificity for gabapentin inhibition of Cav2.1 trafficking.
Gabapentin is well established as an effective treatment for neuropathic pain; however, little is known about its mechanism of action. It binds with high affinity to Ca2+ channel alpha2delta subunits that are expressed in dorsal root ganglia. Mutation of a single alpha2delta amino acid, R217A, eliminates both gabapentin binding and analgesic efficacy. Gabapentin does not seem to have direct Ca2+ channel blocking properties but does affect overall levels of Ca2+channel surface expression in some circumstances. In this report, we examined gabapentin effects on trafficking and voltage-dependent gating properties of recombinant Ca(v)2.1 Ca2+ channel complexes transiently expressed in Xenopus laevis oocytes. We also determined electrophysiologically whether gabapentin causes displacement of beta subunits from Ca(v)2.1 complexes. Our principal findings are as follows: 1) gabapentin inhibits trafficking of recombinant Ca(v)2.1 Ca2+ channels in X. laevis oocytes; 2) gabapentin inhibition occurs in the presence of the Ca2+ channel beta4a subunit but not in the presence of beta4b; 3) gabapentin does not affect Ca(v)2.1 voltage-dependent gating parameters; 4) inhibition of Ca(v)2.1 trafficking is highly dependent on beta-subunit concentration; and 5) gabapentin inhibition of Ca(v)2.1 trafficking can be reversed by the alpha2delta R217A mutation. Overall, our results suggest that gabapentin reduces the number of beta4a-bound Ca(v)2.1 complexes that are successfully trafficked to the plasma membrane. This mechanism may help to explain why gabapentin is both effective and selective in the treatment of neuropathic pain states that involve up-regulation of alpha2delta subunits.


Analysis of illegal peptide drugs via HILIC-DAD-MS.
Biopharmaceuticals have established themselves as highly efficient medicines, and are still one of the fastest growing parts of the health-product industry. Unfortunately, the introduction of these promising new drugs went hand in hand with the creation of a black market for illegal and counterfeit biotechnology drugs. Particularly popular are the lyophilised peptides with a molecular weight of less than 5kDa. Most of them are meant for subcutaneous injection and are easily accessible via the internet. In recent years, different methods based on reversed phase liquid chromatography have been developed to detect and quantify these peptides. The emerging of more polar peptides however requires the introduction of other separation techniques. Therefore, we set out to develop and validate an analytical method based on hydrophilic interaction liquid chromatography (HILIC) to identify and quantify the most frequently encountered illegal peptides on the European market. For this objective, five different HILIC columns were selected and screened for their chromatographic performance. Among those columns, the ZIC HILIC column showed the best performance under the tested screening conditions in terms of resolution and symmetry factor for the targeted peptide set. Hence, the operational conditions were further optimised for the identification of illegal preparations via mass spectrometry (MS) and quantification via UV. Validation was performed via accuracy profiles based on the ISO 17025 guideline. The obtained validated HILIC-method allows for the detection and quantification of the most frequently encountered illegal peptides on the internet in a total run time of 35min including post gradient equilibration and online cleaning step. Combined with a previously developed RPLC-method, the ZIC HILIC system allows for the detection and quantification of a wide spectrum of illicit peptide drugs available on the internet. Furthermore, the developed method could also be envisaged for the detection of new emerging polar peptide drugs.


Electrochemical Generation of Individual Nanobubbles Comprising H2, D2, and HD.
The electrochemical reduction of deuterons (2D+ + 2e- → D2) at Pt nanodisk electrodes (radius = 15-100 nm) in D2O solutions containing deuterium chloride (DCl) results in the formation of a single gas nanobubble at the electrode surface. Analogous to that previously observed for the electrochemical generation of H2 nanobubbles, the nucleation and growth of a stable D2 nanobubble is characterized in voltammetric experiments by a highly reproducible and well-resolved sudden drop in the faradaic current, a consequence of restricted mass transport of D+ to the electrode surface following the liquid-to-gas phase transition. D2 nanobubbles are stable under potential control due to a dynamic equilibrium existing between D2 gas dissolution and electrochemical generation of D2 at the circumference of the Pt nanodisk electrode. Remarkably, within the error of the experimental measurement (<6%), the electrochemical current required to nucleate a D2 gas phase in a D2O solution is identical to that for the H2 gas phase in a H2O solutions, indicating that the concentration required for nucleating a D2 nanobubble in D2O (0.29 M) is ∼1.25 times larger than that for a H2 nanobubble (0.23 M), while the supersaturation is ∼300 in each case. We further demonstrate that individual nanobubbles can be electrogenerated in mixed D2O/H2O solutions containing both D+ and H+ at respective individual concentrations well below those required to nucleate a gas phase containing either pure D2 or H2. This latter finding indicates that the resulting nanobubbles comprise a mixture of D2, H2, and HD molecules with the chemical composition of a nanobubble determined by the concentrations and diffusivities of D+ and H+ in the mixed D2O/H2O solutions.


Origin of the slow afterhyperpolarization and slow rhythmic bursting in striatal cholinergic interneurons.
Striatal cholinergic interneurons recorded in slices exhibit three different firing patterns: rhythmic single spiking, irregular bursting, and rhythmic bursting. The rhythmic single-spiking pattern is governed mainly by a prominent brief afterhyperpolarization (mAHP) that follows single spikes. The mAHP arises from an apamin-sensitive calcium-dependent potassium current. A slower AHP (sAHP), also present in these neurons, becomes prominent during rhythmic bursting or driven firing. Although not apamin sensitive, the sAHP is caused by a calcium-dependent potassium conductance. It is not present after blockade of calcium current with cadmium or after calcium is removed from the media or when intracellular calcium is buffered with bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. It reverses at the potassium equilibrium potential. It can be generated by subthreshold depolarizations and persists after blockade of sodium currents by tetrodotoxin. It is slow, being maximal approximately 1 s after depolarization onset, and takes several seconds to decay. It requires >300-ms depolarizations to become maximally activated. Its voltage sensitivity is sigmoidal, with a half activation voltage of -40 mV. We conclude the sAHP is a high-affinity apamin-insensitive calcium-dependent potassium conductance, triggered by calcium currents partly activated at subthreshold levels. In combination with those calcium currents, it accounts for the slow oscillations seen in a subset of cholinergic interneurons exhibiting rhythmic bursting. In all cholinergic interneurons, it contributes to the slowdown or pause in firing that follows driven activity or prolonged subthreshold depolarizations and is therefore a candidate mechanism for the pause response observed in cholinergic neurons in vivo.


The association between possible stress markers and vocal symptoms.
Stress reaction provokes changes in the body involving cardiovascular alterations, autonomic reactions, neuroendocrine and immunologic as well as psychoneuroimmunologic changes. Both the primary and secondary effect of stress reaction may be of consequence for vocal function. The purpose of this questionnaire study was to determine the effect of stress symptoms on the occurrence of vocal symptoms. The study also aimed at investigating whether a possible effect was different for men and women. A total number of 1728 participants completed a questionnaire concerning speech, language, and voice. Six vocal symptoms and four possible stress symptoms were included in the questionnaire. There was a significant association between stress symptoms and the occurrence of vocal symptoms. The occurrence of muscle tension or a lump in the throat was significant regarding all the four possible stress symptoms. There were also significant results concerning gender difference. All vocal symptoms and two of four stress symptoms were more common among women. Physical changes caused by the stress reaction may result in vocal symptoms as those presented in the study. Both vocal symptoms and stress symptoms were more common among women. Stress should preferably be acknowledged as a risk factor containing and possibly entailing a number of physiological, psychological, and behavioral symptoms affecting the voice negatively.


Confining Ionic Liquids in Developing Quasi-Solid-State Electrolytes for Lithium Metal Batteries.
The concept of confining ionic liquids (ILs) in developing quasi-solid-state electrolytes (QSSEs) has been proposed, where ILs are dispersed in polymer networks/backbones and/or filler/host pores, forming the so-called confinement, and great research progress and promising research results have been achieved. In this review, the progress and achievement in developing QSSEs using IL-confinement for lithium metal batteries (LMBs), together with advanced characterizations and simulations, were surveyed, summarized, and analyzed, where the influence of specific parameters, such as IL (type, content, etc.), substrate (type, structure, surface properties, etc.), confinement methods, and so on, was discussed. The confinement concept was further compared with the conventional one in other research areas. It indicates that the IL-confinement in QSSEs improves the performance of electrolytes, for example, increasing the ionic conductivity, widening the electrochemical window, and enhancing the cycle performance of the assembled cells, and being different from those in other areas, that is, the IL-confinement concept in the battery area is in a broad extent. Finally, insights into developing QSSEs in LMBs with the confinement strategy were provided to promote the development and application of QSSE LMBs.


[Determination of retention during phacoemulsification with AOL in patients with open-angle glaucoma].
Phacoemulsification is the most sparing technique of cataract removal in patients with open-angle glaucoma. The retention of intraocular fluid should be determined to choose a pathogenetically oriented antiglaucoma operation for uncompensated open-angle glaucoma (OAG). The study of the Schlemm's canal retrograde blood filling phenomenon may be a technique of the most informative value. The phenomenon was studied on 102 eyes (in 82 cases of glaucoma) during phacoemulsification, which indicated the high effectiveness and safety of the use of this intraoperative method. Its advantage is that manipulations are easy-to-use and accessible. This method is of particular value in treating patients with cataract concurrent with OAG, who are to undergo phacoemulsification with a one-stage pathogenetically oriented antiglaucoma operation.


Time Discounting and Credit Market Access in a Large-Scale Cash Transfer Programme.
Time discounting is thought to influence decision-making in almost every sphere of life, including personal finances, diet, exercise and sexual behavior. In this article we provide evidence on whether a national poverty alleviation program in Kenya can affect inter-temporal decisions. We administered a preferences module as part of a large-scale impact evaluation of the Kenyan Government's Cash Transfer for Orphans and Vulnerable Children. Four years into the program we find that individuals in the treatment group are only marginally more likely to wait for future money, due in part to the erosion of the value of the transfer by inflation. However among the poorest households for whom the value of transfer is still relatively large we find significant program effects on the propensity to wait. We also find strong program effects among those who have access to credit markets though the program itself does not improve access to credit.


First Video Case Report of Chronic Retrograde Jejunojejunal Intussusception after Subtotal Gastrectomy with Braun's Anastomosis.
Intussusception, which is seen rarely in adults, is defined as the pulling or invagination of a part of the intestine into another segment of the intestine. In this case report we present chronic retrograde jejunojejunal intussusception following gastric surgery with Braun's anastomosis in adult with video presentation. A 66-year-old woman, who had undergone gastric surgery 39 years ago and cholecystectomy 20 years ago, was admitted to our clinic with the complaints about weight loss, abdominal pain, nausea, and vomiting. Upper gastrointestinal endoscopy (UGISE) was applied, and patient was treated with surgery. This case report indicates that intussusception should be considered in the presence of clinical complaints following gastric surgery, as well as importance of endoscopy in diagnosis.


Rhabdomyolysis associated with Mycoplasma pneumoniae pneumonia.
We report a case of rhabdomyolysis associated with Mycoplasma pneumoniae pneumonia in a 37-year-old Sri Lankan man who presented to the emergency department with complaints of feverishness, shortness of breath, cough, and generalised muscle pain. He had a serum creatinine kinase of 14 220 U/L, serum myoglobin of 1822 ng/mL, and serum creatinine of 195 µmol/L. His chest X-ray revealed bibasilar interstitial infiltrates. The antimycoplasma antibody titre was high. The patient was successfully treated with aggressive intravenous fluid replacement and azithromycin. The outcome was rapidly favourable, allowing us to discharge the patient 12 days after admission. On discharge, the serum creatinine kinase was 924 U/L and the creatinine was 126 µmol/L; the chest examination was unremarkable.


Pan-American League of Associations for Rheumatology (PANLAR) recommendations and guidelines for musculoskeletal ultrasound training in the Americas for rheumatologists.
To develop guidelines for Musculoskeletal Ultrasound (MSKUS) training for rheumatologists in the Americas. A total of 25 Rheumatologists from 19 countries of the American Continent participated in a consensus-based interactive process (Delphi method) using 2 consecutive electronic questionnaires. The first questionnaire included the following: the relevance of organizing courses to teach MSKUS to Rheumatologists, the determination of the most effective educational course models, the trainee levels, the educational objectives, the requirements for passing the course(s), the course venues, the number of course participants per instructor, and the percentage of time spent in hands-on sessions. The second questionnaire consisted of questions that did not achieve consensus (>65%) in the first questionnaire, topics, and pathologies to be covered at each course MSKUS level. General consensus was obtained for MSKUS courses to be divided into 3 educational levels: basic, intermediate, and advanced. These courses should be taught using a theoretical-didactic and hands-on model. In addition, the group established the minimum requirements for attending and passing each MSKUS course level, the ideal number of course participants per instructor (4 participants/instructor), and the specific topics and musculoskeletal pathologies to be covered. In the same manner, the group concluded that 60% to 70% of course time should be focused on hands-on sessions. A multinational group of MSKUS sonographers using a consensus-based questionnaire (Delphi method) established the first recommendations and guidelines for MSKUS course training in the Americas. Pan-American League of Associations for Rheumatology urges that these guidelines and recommendations be adopted in the future by both national and regional institutions in the American continent involved in the training of Rheumatologists for the performance of MSKUS.


Managing high-risk patients: the Mass General care management programme.
The Massachusetts General Care Management Program (Mass General CMP or CMP) was designed as a federally supported demonstration to test the impact of intensive, practice-based care management on high-cost Medicare fee-for-service (FFS) beneficiaries-primarily older persons-with multiple hospitalisations and multiple chronic conditions. The Massachusetts General Care Management Program operated over a 6-year period in two phases (3 years each). It started during the first phase at Massachusetts General Hospital, a major academic medical centre in Boston, Massachusetts in collaboration with Massachusetts General Physicians Organisation. During the second phase, the programme expanded to two more affiliated sites in and around the Boston area, including a community hospital, as well as incorporated several modifications primarily focused on the management of transitions to post-acute care in skilled nursing facilities. At the close of the demonstration in July 2012, Mass General Massachusetts General Care Management Program became a component of a new Pioneer accountable care organisation (ACO). The Massachusetts General Care Management Program is focused on individuals meeting defined eligibility criteria who are offered care that is integrated by a case manager embedded in a primary care practice. The demonstration project showed substantial cost savings compared to fee-for-service patients served in the traditional Medicare system but no impact on hospital readmissions. The Massachusetts General Care Management Program does not rest upon a "whole systems" approach to integrated care. It is an excellent example of how an innovative care co-ordination programme can be implemented in an existing health-care organisation without making fundamental changes in its underlying structure or the way in which direct patient care services are paid for. The accountable care organisation version of the Massachusetts General Care Management Program includes the staffing structure, standards of practice, collaborative approach to care transitions and information technology tools that were used in the original demonstration project.


Leptin release is decreased in white adipocytes isolated from progesterone-treated rats.
Previously, it has been proposed that progesterone has an inhibitory effect on leptin secretion by white adipocytes, because female rats treated with progesterone show unchanged plasma leptin concentrations despite heavier fat depots. In this study, we show that adipocytes isolated from intact rats release the same amount of leptin either in the presence or the absence of progesterone in the incubation medium. However, when we isolated white adipocytes from progesterone-treated and sham-treated rats and measured their leptin release for 6 hr, we found that adipocytes isolated from rats treated with progesterone for 72 hr showed a lower leptin release than those of sham-treated rats. These results confirm the proposed inhibitory action of progesterone on leptin production.


Epigenome-wide association study of mitochondrial genome copy number.
We conducted cohort- and race-specific epigenome-wide association analyses of mitochondrial deoxyribonucleic acid (mtDNA) copy number (mtDNA CN) measured in whole blood from participants of African and European origins in five cohorts (n = 6182, mean age = 57-67 years, 65% women). In the meta-analysis of all the participants, we discovered 21 mtDNA CN-associated DNA methylation sites (CpG) (P < 1 × 10-7), with a 0.7-3.0 standard deviation increase (3 CpGs) or decrease (18 CpGs) in mtDNA CN corresponding to a 1% increase in DNA methylation. Several significant CpGs have been reported to be associated with at least two risk factors (e.g. chronological age or smoking) for cardiovascular disease (CVD). Five genes [PR/SET domain 16, nuclear receptor subfamily 1 group H member 3 (NR1H3), DNA repair protein, DNA polymerase kappa and decaprenyl-diphosphate synthase subunit 2], which harbor nine significant CpGs, are known to be involved in mitochondrial biosynthesis and functions. For example, NR1H3 encodes a transcription factor that is differentially expressed during an adipose tissue transition. The methylation level of cg09548275 in NR1H3 was negatively associated with mtDNA CN (effect size = -1.71, P = 4 × 10-8) and was positively associated with the NR1H3 expression level (effect size = 0.43, P = 0.0003), which indicates that the methylation level in NR1H3 may underlie the relationship between mtDNA CN, the NR1H3 transcription factor and energy expenditure. In summary, the study results suggest that mtDNA CN variation in whole blood is associated with DNA methylation levels in genes that are involved in a wide range of mitochondrial activities. These findings will help reveal molecular mechanisms between mtDNA CN and CVD.


Hegemony, emancipation and mythology.
This epilogue offers a critical overview of some current interpretations of modern sport's origins and evolution in Latin America by certain European and North American commentators. These analyses still adhere to the tradition of the New World's creation. They seldom refer to primary sources. Conclusions are often born out of patronizing compassion and exotic impression. Latin American sources, if any, are mostly quoted when they serve a pessimistic historical bias cultivated by outsiders. In fact, Latin American sport represents a mirror image of more developed areas and indeed a reinvention of sport in specific cases of community isolation and self-development.


Bcl-3: A Double-Edged Sword in Immune Cells and Inflammation.
The NF-κB transcription factor family controls the transcription of many genes and regulates a number of pivotal biological processes. Its activity is regulated by the IκB family of proteins. Bcl-3 is an atypical member of the IκB protein family that regulates the activity of nuclear factor NF-κB. It can promote or inhibit the expression of NF-κB target genes according to the received cell type and stimulation, impacting various cell functions, such as proliferation and differentiation, induction of apoptosis and immune response. Bcl-3 is also regarded as an environment-dependent cell response regulator that has dual roles in the development of B cells and the differentiation, survival and proliferation of Th cells. Moreover, it also showed a contradictory role in inflammation. At present, in addition to the work aimed at studying the molecular mechanism of Bcl-3, an increasing number of studies have focused on the effects of Bcl-3 on inflammation, immunity and malignant tumors in vivo. In this review, we focus on the latest progress of Bcl-3 in the regulation of the NF-κB pathway and its extensive physiological role in inflammation and immune cells, which may help to provide new ideas and targets for the early diagnosis or targeted treatment of various inflammatory diseases, immunodeficiency diseases and malignant tumors.


Processes affecting the dissipation of the herbicide isoxaflutole and its diketonitrile metabolite in agricultural soils under field conditions.
Two-year field dissipation studies were conducted in three soil types in Minnesota to examine the processes affecting the dissipation of the herbicide isoxaflutole and its phytotoxic diketonitrile metabolite (DKN) under relatively cool, wet soil conditions. Plots of cuphea were treated with isoxaflutole and potassium bromide, a nonsorbed, nondegraded tracer. Replicate soil cores were collected six times during the growing season to a depth of 1 m, and the bromide or herbicide concentration was measured in each of five depth increments. The dissipation half-life (DT50) of isoxaflutole + DKN was 8-18 days in each soil. Bromide and herbicide concentrations were low at depths >40 cm throughout the study, and herbicide concentrations in soil 100 days after application were usually undetectable. Simulation modeling using Hydrus-1D for the loam soil suggested that plant uptake was an important mechanism of dissipation.


The development of a 'wet leg' pathway for chronic oedema.
Chronic oedema and leaking lymphorrhoea can be distressing and challenging for patients to live with. This article introduces the Chronic Oedema 'Wet Leg' Pathway, which became an output from the Lymphoedema Network Wales 'On the Ground Education Project' ( Thomas et al, 2017 ). The evidence-based pathway provides nurses and therapists with clear guidelines to support prompt, efficient and effective management of all patients with chronic oedema and wet legs. This article presents the evidence and demonstrates the benefits of timely active treatment for all patients with leaking lymphorrhoea, which includes decreasing pain and improving mobility and daily activities. Other benefits include reducing the risk of cellulitis and the development of chronic wounds. The pathway also demonstrates opportunities for the NHS by decreasing both the number of community nursing visits and the number of wound dressings used. By actively treating symptoms-rather than simply mopping up the leaking lymphorrhoea-this pathway demonstrates value-based health care by reducing waste, harm and variation within the NHS.


Reactivity of Tantalum Carbide Cluster Anions TaC n- ( n = 1-4) with Dinitrogen.
Dinitrogen activation/fixation is one of the most important and challenging subjects in synthetic as well as theoretical chemistry. In this study, the adsorption reactions of N2 onto TaC n- ( n = 1-4) cluster anions have been investigated by means of mass spectrometry in conjunction with density functional theory calculations. Following the experimental results that only TaC4- was observed to adsorb N2, theoretical calculations predicted that the TaC4- reaction system (TaC4- + N2 → TaC4N2-) has a negligible barrier on the approach of N2 molecule while insurmountable barriers are located on the reaction pathways of TaC1-3-/N2 reaction systems. The natural bond orbital and molecular orbital analysis indicated that the more positive charge on the metal center of TaC1-4- would facilitate the initial approach of the nonpolar N2 molecule, and the appropriate frontier orbital of TaC1-4- with proper symmetry (π-type 5d orbital) which can match up well with the π* antibonding orbital of the N2 molecule with less σ repulsion and more possibility for π back-donation would be helpful for the formation of the stable encounter complexes. This study reveals the fundamental rules and key factors governing the N2 adsorption.


Chemical regulation of carbon quantum dots from synthesis to photocatalytic activity.
Carbon quantum dots (CQDs) were synthesized by heating various carbon sources in HNO3 solution at reflux, and the effects of HNO3 concentration on the size of the CQDs were investigated. Furthermore, the oxygen-containing surface groups of as-prepared CQDs were selectively reduced by NaBH4 , leading to new surface states. The experimental results show that the sizes of CQDs can be tuned by HNO3 concentration and then influence their photoluminescent behaviors; the photoluminescent properties are related to both the size and surface state of the CQDs, but the photocatalytic activities are determined by surface states alone. The different oxygen-containing groups on the surface of the CQDs can induce different degrees of the band bending upward, which determine the separation and combination of the electron-hole pairs. The high upward band bending, which is induced by C=O and COOH groups, facilitates separation of the electron-hole pairs and then enhances high photocatalytic activity. In contrast, the low upward band bending induced by C-OH groups hardly prevents the electron-hole pairs from surface recombination and then exhibits strong photoluminescence. Therefore, both the photocatalytic activities and optical properties of CQDs can be tuned by their surface states.


Predicting delayed cerebral ischemia after subarachnoid hemorrhage using physiological time series data.
To develop and validate a prediction model for delayed cerebral ischemia (DCI) after subarachnoid hemorrhage (SAH) using a temporal unsupervised feature engineering approach, demonstrating improved precision over standard features. 488 consecutive SAH admissions from 2006 to 2014 to a tertiary care hospital were included. Models were trained on 80%, while 20% were set aside for validation testing. Baseline information and standard grading scales were evaluated: age, sex, Hunt Hess grade, modified Fisher Scale (mFS), and Glasgow Coma Scale (GCS). An unsupervised approach applying random kernels was used to extract features from physiological time series (systolic and diastolic blood pressure, heart rate, respiratory rate, and oxygen saturation). Classifiers (Partial Least Squares, linear and kernel Support Vector Machines) were trained on feature subsets of the derivation dataset. Models were applied to the validation dataset. The performances of the best classifiers on the validation dataset are reported by feature subset. Standard grading scale (mFS): AUC 0.58. Combined demographics and grading scales: AUC 0.60. Random kernel derived physiologic features: AUC 0.74. Combined baseline and physiologic features with redundant feature reduction: AUC 0.77. Current DCI prediction tools rely on admission imaging and are advantageously simple to employ. However, using an agnostic and computationally inexpensive learning approach for high-frequency physiologic time series data, we demonstrated that our models achieve higher classification accuracy.


[Health economic evaluation of community-based psychiatric interventions].
The health economic evaluation of community mental health interventions can contribute to an optimization of the allocation of mental health care resources. Existing studies on the health economic evaluation of community mental health interventions include a wide range of treatment options from the deinstitutionalization of long-term inpatients into community-based services to specialized services for specific patient groups. The results of these studies indicate that community-based mental health care is not generally less costly in comparison to more institutionalized forms of care. Programs that are based on the ACT approach have been found to be more efficient than inpatient care. Compared to the current standard of mental health care in most western European countries, an intensification of community mental health interventions could increase the efficiency of psychiatric treatment especially for heavy users and first episode patients. However, methodological flaws and a lack of national studies limit the validity of current health economic investigations, particularly for Germany.


Noninvasive prenatal screening for fetal common sex chromosome aneuploidies from maternal blood.
Objective To explore the feasibility of high-throughput massively parallel genomic DNA sequencing technology for the noninvasive prenatal detection of fetal sex chromosome aneuploidies (SCAs). Methods The study enrolled pregnant women who were prepared to undergo noninvasive prenatal testing (NIPT) in the second trimester. Cell-free fetal DNA (cffDNA) was extracted from the mother's peripheral venous blood and a high-throughput sequencing procedure was undertaken. Patients identified as having pregnancies associated with SCAs were offered prenatal fetal chromosomal karyotyping. Results The study enrolled 10 275 pregnant women who were prepared to undergo NIPT. Of these, 57 pregnant women (0.55%) showed fetal SCA, including 27 with Turner syndrome (45,X), eight with Triple X syndrome (47,XXX), 12 with Klinefelter syndrome (47,XXY) and three with 47,XYY. Thirty-three pregnant women agreed to undergo fetal karyotyping and 18 had results consistent with NIPT, while 15 patients received a normal karyotype result. The overall positive predictive value of NIPT for detecting SCAs was 54.54% (18/33) and for detecting Turner syndrome (45,X) was 29.41% (5/17). Conclusion NIPT can be used to identify fetal SCAs by analysing cffDNA using massively parallel genomic sequencing, although the accuracy needs to be improved particularly for Turner syndrome (45,X).


Rapid Separation of Asiatic Acid, Quercetin, and Kaempferol from Traditional Chinese Medicine Centella asiatica (L.) Urban Using HSCCC-Semi-Prep-HPLC and the Assessment of Their Potential as Fatty Acid Synthase Inhibitors.
The main objective of this study was to rapidly separate asiatic acid (AA), quercetin (QCN), and kaempferol (KPL) from Centella asiatica (L.) Urban using high-speed counter-current chromatography (HSCCC) in tandem with the UV detector of semipreparative high-performance liquid chromatography (Semi-Prep-HPLC) and to evaluate their potential as inhibitors of fatty acid synthetase (FAS). To efficiently prepare large amounts of AA, QCN, and KPL from Centella asiatica (L.) Urban, rapid and simple methods by HSCCC were established respectively based on the partition coefficients (K values) of crude samples. The conditions of HSCCC-Semi-Prep-HPLC for the large-scale separation of AA, QCN, and KPL from Centella asiatica (L.) Urban were established and optimized. This included selecting the solvent system, flow rate, rotation speed, and so on. HSCCC-Semi-Prep-HPLC was successfully applied to separate and purify AA, QCN, and KPL, with n-hexane-n-butanol-methanol-water (3 : 1 : 3 : 3, V : V : V : V) as the solvent system for AA, which was detected at a wavelength of 210 nm with the stationary phase retention of 70%, and with n-hexane-ethyl acetate-methanol-water (0.8 : 0.9 : 1.2 : 1, V : V : V : V) as the solvent system for the co-separation of QCN and KPL, which was detected at a wavelength of 254 nm with the stationary phase retention of 65%. AA could be isolated at a large scale with high purity (>91.0%) in only one-step HSCCC-Semi-Prep-HPLC separation (within 150 min) under the optimized conditions. Meanwhile, QCN and KPL could be simultaneously isolated at a large scale with high purity (>99.1%) by another one-step HSCCC-Semi-Prep-HPLC separation (within 240 min) under the optimized conditions. The assessment of inhibition potential revealed that AA exhibited the strongest inhibitory effect on FAS, with an IC50 of 9.52 ± 0.76 μg/mL. Madecassic acid (MA) followed closely with IC50 values of 10.84 ± 0.92 μg/mL. QCN and KPL showed similar and relatively weaker inhibitory effects on FAS, with IC50 values of 43.09 ± 2.98 μg/mL and 36.90 ± 1.83 μg/mL, respectively. Overall, the HSCCC-Semi-Prep-HPLC method proved to be a highly efficient and reliable technique for separating AA, QCN, and KPL from Centella asiatica (L.) Urban, and the isolated compounds showed potential as FAS inhibitors.


Representations of nurses and individuals in psychological distress in the photographic exhibit USAnatomy, by Steven Klein.
Objectives To analyze the representations of the nurse and individual in mental suffering portrayed in the photographic work by Steven Klein, in the USAnatomy exhibit held at the Museum of Sculpture (Museu da escultura) in São Paulo, in 2011. Methods Qualitative study carried out in 2012. Three photographs were submitted to iconographic analysis. The interpretation of the findings was based on theoretical frameworks of Foucault and Bourdieu on power relations. Results The nurse is represented as a sensual, insensitive person, with the power to control and torture while the person in psychological distress is represented as dirty, imprisoned and subjected to an asymmetrical relationship of power with the nurse. Final considerations Relationships of submission and symbolic dominance, in which the person in psychological distress has their body molded by discipline imposed by the nurse. Stereotypes of the image of the nurse oppose the ideology of the profession, which is to ensure the integrity of those being cared for.


A simple high-dose gentamicin regimen showed no side effects among neonates.
Treatment of infections in neonates with gentamicin is a balance between optimising bactericidal effect and minimising adverse effects. Previously, at the Neonatal Intensive Care Unit (NICU) at Kolding Hospital, Denmark, neonates suspected of having infections were treated daily with gentamicin 5 mg/kg for the first three days, thus exposing the smallest neonates to double gentamicin amounts compared with those used in most Danish NICUs. We aimed to evaluate if this regimen increased the trough values and oto- and nephrotoxicity. Neonates admitted to the NICU between 2008 and 2012 and treated with gentamicin were included retrospectively in the study. Neonates with trough serum (S)-gentamicin level ≥ 2.0 mg/l before the third dose were reviewed in detail. In total, S-gentamicin level was measured in 253 treated neonates of whom 7% displayed elevated trough values. Neonates < 32 weeks of age had a slightly higher incidence of S-gentamicin level ≥ 2.0 mg/l compared with less premature and mature infants (16%, 13%, and 2%, respectively). No oto- or nephrotoxicity was found despite the high-dose gentamicin regimen. The incidence of elevated S-gentamicin trough levels was increased among very premature neonates. However, no evidence of oto- or nephrotoxicity was observed. This simple regimen of gentamicin 5 mg/kg for the first three days should be considered for all neonates as it potentially minimises the risk of dosing errors and bacterial breakthrough infection. none. Danish Data Protection Agency (2008-58-0035).


Long-term Outcome after Thrombolysis for Acute Lower Limb Ischaemia.
The purpose was to study long-term outcome after thrombolysis for acute arterial lower limb ischaemia, and to evaluate the results depending on the underlying aetiology of arterial occlusion. This was a retrospective study of patients entered into a prospective database. Patients were identified in prospective databases from two vascular centres, including a large number of variables. Case records were analysed retrospectively. Through cross linkage with the Population Registry 100% accurate survival data were obtained. Between January 2001 and December 2013, 689 procedures were included. The aetiology of ischaemia was graft/stent/stent graft occlusion in 39.8%, arterial thrombosis in 27.7%, embolus in 25.1% and popliteal aneurysm in 7.4%. The mean follow-up was 59.4 months (95% CI, 56.1-62.7), during which 32.9% needed further re-interventions, 16.4% underwent amputation without re-intervention, and 50.7% had no re-intervention. The need for re-intervention during follow-up was 48.0% in the graft/stent occlusions group, 34.0% of the popliteal aneurysm group, 25.4% in the thrombosis group, and 16.3% in the embolus group (p < .001). The overall primary patency rates were 69.1% and 55.9% at 1 and 5 years, respectively. Primary patency at 5 years was higher for the embolus group (83.3%, p = .002) and lower for the occluded graft/stent group (43.3%, p < .001). Secondary patency rates were 80.1% and 75.2% at 1 and 5 years, respectively, without difference between the subgroups. The amputation rate was lower in the embolic group at 1 and 5 years (8.1% and 11.1%, respectively, p = .001). Survival was higher in the group with occluded popliteal aneurysms at 5 years (83.3%, p = 0.004). Amputation free survival was 72.1% and 45.2% at 1 and 5 years; lower in the occluded graft/stent group at five years (37.9%, p = .007). Intra-arterial thrombolytic therapy achieves good medium and long-term clinical outcome, reducing the need of open surgical treatment in most patients.