Saccharomyces boulardii inhibits ERK1/2 mitogen-activated protein kinase activation both in vitro and in vivo and protects against Clostridium difficile toxin A-induced enteritis.
Saccharomyces boulardii (Sb), a probiotic yeast, protects against intestinal injury and inflammation caused by a wide variety of enteric pathogens, including Clostridium difficile. Given the broad range of protective effects of Sb in multiple gastrointestinal disorders, we hypothesize that Sb modulates host signaling pathways involved in intestinal inflammatory responses. In this study, we found that Sb culture supernatant (SbS) inhibits interleukin-8 production induced by C. difficile toxin A or IL-1beta in human colonocyte NCM460 cells in a dose-dependent fashion. Furthermore, SbS inhibited IL-1beta and toxin A induced Erk1/2 and JNK/SAPK but not p38 activation in NCM460 cells. To test whether this inhibition also occurs in vivo, we used a previously established mouse ileal loop model. On its own, SbS had no significant effect on basal fluid secretion or intestinal histology. However, Erk1/2 activation was significantly inhibited by SbS in toxin A exposed mouse ileal mucosa. In control loops, toxin A increased fluid secretion (2.2-fold), histological score (3.3-fold), and levels of the chemokine KC (4.5-fold). SbS pretreatment completely normalized toxin A mediated fluid secretion (p < 0.01), and histopathologic changes (p < 0.01) and substantially inhibited toxin A-associated KC increases (p < 0.001). In summary, the probiotic yeast S. boulardii inhibits C. difficile toxin A-associated enteritis by blocking the activation of Erk1/2 MAP kinases. This study indicates a new mechanism whereby Sb protects against intestinal inflammation and supports the hypothesis that Sb modulates host inflammatory signaling pathways to exert its beneficial effects.


Numerical Method of Characteristics for One-Dimensional Blood Flow.
Mathematical modeling at the level of the full cardiovascular system requires the numerical approximation of solutions to a one-dimensional nonlinear hyperbolic system describing flow in a single vessel. This model is often simulated by computationally intensive methods like finite elements and discontinuous Galerkin, while some recent applications require more efficient approaches (e.g. for real-time clinical decision support, phenomena occurring over multiple cardiac cycles, iterative solutions to optimization/inverse problems, and uncertainty quantification). Further, the high speed of pressure waves in blood vessels greatly restricts the time step needed for stability in explicit schemes. We address both cost and stability by presenting an efficient and unconditionally stable method for approximating solutions to diagonal nonlinear hyperbolic systems. Theoretical analysis of the algorithm is given along with a comparison of our method to a discontinuous Galerkin implementation. Lastly, we demonstrate the utility of the proposed method by implementing it on small and large arterial networks of vessels whose elastic and geometrical parameters are physiologically relevant.


[Molecular dating in the genomic era].
The comparison of DNA and protein sequences of extant species might be informative for reconstructing the chronology of evolutionary events on Earth. A phylogenetic tree inferred from molecular data directly depicts the evolutionary affinities of species and indirectly allows estimating the age of their origin and diversification. Molecular dating is achieved by assuming the molecular clock hypothesis, i.e., that the rate of change of nucleotide and amino acid sequences is on average constant over geological time. If paleontological calibrations are available, then absolute divergence times of species can be estimated. However, three major difficulties potentially hamper molecular dating : (1) a limited sample of genes and organisms, (2) a limited number of fossil references, and (3) pervasive variations of molecular evolutionary rates among genomes and species. To circumvent these problems, different solutions have been recently proposed. Larger data sets are built with more genes and more species sampled through the mining of an increasing number of genomes. Moreover, independent key fossils are identified to calibrate molecular clocks, and the uncertainty on their age is integrated in subsequent analyses. Finally, models of molecular rate variations are constructed, and incorporated in the so-called relaxed molecular clock approaches. As an illustration of these improvements, we mention that the debated age of the animal (bilaterian metazoans) diversification may have occurred between 642-761 million years ago (Mya), roughly 100 Ma before the Cambrian explosion. Among mammals, the initial diversification of major placental groups may have taken place around 100 Mya, well before the Cretaceous/Tertiary boundary marking the extinction of dinosaurs.


Dual Inhibition of Key Proliferation Signaling Pathways in Triple-Negative Breast Cancer Cells by a Novel Derivative of Taiwanin A.
The treatment of breast cancer cells obtained by blocking the aberrant activation of the proliferation signaling pathways PI3K/Akt/mTOR and MEK/ERK has received considerable attention in recent years. Previous studies showed that Taiwanin A inhibited the proliferation of several types of cancer cells. In this study, we report that 3,4-bis-3,4,5-trimethoxybenzylidene-dihydrofuran (BTMB), a novel derivative of Taiwanin A, significantly inhibited the proliferation of triple-negative breast cancer (TNBC) cells both in vitro and in vivo The results show that BTMB inhibited the proliferation of human TNBC cells by the induction of cell-cycle arrest and apoptosis in a dose-dependent fashion. BTMB inhibited the expression of β-catenin, cdc2 and the cell-cycle regulatory proteins, cyclin A, cyclin D1, and cyclin E. The mechanism of action was associated with the suppression of cell survival signaling through inactivation of the Akt and ERK1/2 signaling pathways. Moreover, BTMB induced cell apoptosis through an increase in the expression of BAX, cleaved caspase-3, and cleaved PARP. Moreover, BTMB inhibited TNBC cell colony formation and sensitized TNBC cells to cisplatin, a chemotherapeutic drug. In a TNBC mouse xenograft model, BTMB significantly inhibited the growth of mammary carcinomas through decreased expression of cyclin D1. BTMB was shown to significantly suppress the growth of mammary carcinoma and therefore to have potential as an anticancer therapeutic agent. Mol Cancer Ther; 16(3); 480-93. ©2016 AACR.


Phosphate Groups in the Lipid A Moiety Determine the Effects of LPS on Hepatic Stellate Cells: A Role for LPS-Dephosphorylating Activity in Liver Fibrosis.
Alkaline phosphatase (AP) activity is highly upregulated in plasma during liver diseases. Previously, we demonstrated that AP is able to detoxify lipopolysaccharide (LPS) by dephosphorylating its lipid A moiety. Because a role of gut-derived LPS in liver fibrogenesis has become evident, we now examined the relevance of phosphate groups in the lipid A moiety in this process. The effects of mono-phosphoryl and di-phosphoryl lipid A (MPLA and DPLA, respectively) were studied in vitro and LPS-dephosphorylating activity was studied in normal and fibrotic mouse and human livers. The effects of intestinal AP were studied in mice with CCL4-induced liver fibrosis. DPLA strongly stimulated fibrogenic and inflammatory activities in primary rat hepatic stellate cells (rHSCs) and RAW264.7 macrophages with similar potency as full length LPS. However, MPLA did not affect any of the parameters. LPS-dephosphorylating activity was found in mouse and human livers and was strongly increased during fibrogenesis. Treatment of fibrotic mice with intravenous intestinal-AP significantly attenuated intrahepatic desmin+- and αSMA+ -HSC and CD68+- macrophage accumulation. In conclusion, the lack of biological activity of MPLA, contrasting with the profound activities of DPLA, shows the relevance of LPS-dephosphorylating activity. The upregulation of LPS-dephosphorylating activity in fibrotic livers and the protective effects of exogenous AP during fibrogenesis indicate an important physiological role of intestinal-derived AP during liver fibrosis.


Solid organ transplantation in primary mitochondrial disease: Proceed with caution.
Solid organ transplants are rarely performed in both adult and pediatric patients with primary mitochondrial disease. Poor outcomes have been described in case reports and small case series. It is unclear whether the underlying genetic disease has a significant impact on post-transplant morbidity and mortality. Data were obtained for 35 patients from 17 Mitochondrial Disease Centers across North America, the United Kingdom and Australia. Patient outcomes were noted after liver, kidney or heart transplantation. Excluding patients with POLG-related disease, post-transplant survival approached or met outcomes seen in non-mitochondrial disease transplant patients. The majority of mitochondrial disease patients did not have worsening of their mitochondrial disease within 90-days post-transplant. Post-transplant complications, including organ rejection, were not a common occurrence and were generally treatable. Many patients did not have a mitochondrial disease considered or diagnosed prior to transplantation. In conclusion, patients with mitochondrial disease in this cohort generally tolerated solid-organ transplantation. Such patients may not need to be excluded from transplant solely for their mitochondrial diagnosis; additional caution may be needed for patients with POLG-related disease. Transplant teams should be aware of mitochondrial disease as an etiology for organ-failure and consider appropriate consultation in patients without a known cause of their symptoms.


The site specific demethylation in the 5'-regulatory area of NMDA receptor 2B subunit gene associated with CIE-induced up-regulation of transcription.
The NMDA receptor represents a particularly important site of ethanol action in the CNS. We recently reported that NMDA receptor 2B (NR2B) gene expression was persistently up-regulated following chronic intermittent ethanol (CIE) treatment. Increasing evidence that epigenetic mechanisms are involved in dynamic and long-lasting regulation of gene expression in multiple neuroadaptive processes prompted us to investigate the role of DNA methylation in mediating CIE-induced up-regulation of NR2B gene transcription. To dissect the changes of DNA methylation in the NR2B gene, we have screened a large number of CpG sites within its 5'-regulatory area following CIE treatment. Primary cortical cultured neurons were subjected to ethanol treatment in a CIE paradigm. Bisulfite conversion followed by pyrosequencing was used for quantitative measurement and analysis of CpG methylation status within the 5'-regulatory area of the NR2B gene; chromatin immunoprecipitation (ChIP) assay was used to examine DNA levels associated with methylation and transcription factor binding. Electrophoretic mobility shift assay (EMSA) and in vitro DNA methylation assays were performed to determine the direct impact of DNA methylation on the interaction between DNA and transcription factor and promoter activity. Analysis of individual CpG methylation sites within the NR2B 5'regulatory area revealed three regions with clusters of site-specific CpG demethylation following CIE treatment and withdrawal. This was confirmed by ChIP showing similar decreases of methylated DNA in the same regions. The CIE-induced demethylation is characterized by being located near certain transcription factor binding sequences, AP-1 and CRE, and occurred during treatment as well as after ethanol withdrawal. Furthermore, the increase in vitro of methylated DNA decreased transcription factor binding activity and promoter activity. An additional ChIP assay indicated that the CIE-induced DNA demethylation is accompanied by increased occupation by transcription factors. These results suggest an important role of DNA demethylation in mediating CIE-induced NR2B gene up-regulation, thus implicating a novel molecular site of alcohol action.


Breast cancer dormancy can be maintained by small numbers of micrometastases.
Late relapse of breast cancer can occur more than 25 years after primary diagnosis. During the intervening years between initial treatment and relapse, occult cancers are maintained in an apparent state of dormancy that is poorly understood. In this study, we applied a probabilistic mathematical model to long-term follow-up studies of postresection patients to investigate the factors involved in mediating breast cancer dormancy. Our results suggest that long-term dormancy is maintained most often by just one growth-restricted dangerous micrometastasis. Analysis of the empirical data by Approximate Bayesian Computation indicated that patients in dormancy have between 1 and 5 micrometastases at 10 years postresection, when they escape growth restriction with a half-life of <69 years and are >0.4 mm in diameter. Before resection, primary tumors seed at most an average of 6 dangerous micrometastases that escape from growth restriction with a half-life of at least 12 years. Our findings suggest that effective preventive treatments will need to eliminate these small numbers of micrometastases, which may be preangiogenic and nonvascularized until they switch to growth due to one oncogenic mutation or tumor suppressor gene inactivation. In summary, breast cancer dormancy seems to be maintained by small numbers of sizeable micrometastases that escape from growth restriction with a half-life exceeding 12 years.


Covid-19 Transmission Trajectories-Monitoring the Pandemic in the Worldwide Context.
The Covid-19 pandemic is developing worldwide with common dynamics but also with marked differences between regions and countries. These are not completely understood, but presumably, provide a clue to find ways to mitigate epidemics until strategies leading to its eradication become available. We describe an iteractive monitoring tool available in the internet. It enables inspection of the dynamic state of the epidemic in 187 countries using trajectories that visualize the transmission and removal rates of the epidemic and in this way bridge epi-curve tracking with modelling approaches. Examples were provided which characterize state of epidemic in different regions of the world in terms of fast and slow growing and decaying regimes and estimate associated rate factors. The basic spread of the disease is associated with transmission between two individuals every two-three days on the average. Non-pharmaceutical interventions decrease this value to up to ten days, whereas 'complete lock down' measures are required to stop the epidemic. Comparison of trajectories revealed marked differences between the countries regarding efficiency of measures taken against the epidemic. Trajectories also reveal marked country-specific recovery and death rate dynamics. The results presented refer to the pandemic state in May to July 2020 and can serve as 'working instruction' for timely monitoring using the interactive monitoring tool as a sort of 'seismometer' for the evaluation of the state of epidemic, e.g., the possible effect of measures taken in both, lock-down and lock-up directions. Comparison of trajectories between countries and regions will support developing hypotheses and models to better understand regional differences of dynamics of Covid-19.


Olfaction written in bone: cribriform plate size parallels olfactory receptor gene repertoires in Mammalia.
The evolution of mammalian olfaction is manifested in a remarkable diversity of gene repertoires, neuroanatomy and skull morphology across living species. Olfactory receptor genes (ORGs), which initiate the conversion of odorant molecules into odour perceptions and help an animal resolve the olfactory world, range in number from a mere handful to several thousand genes across species. Within the snout, each of these ORGs is exclusively expressed by a discrete population of olfactory sensory neurons (OSNs), suggesting that newly evolved ORGs may be coupled with new OSN populations in the nasal epithelium. Because OSN axon bundles leave high-fidelity perforations (foramina) in the bone as they traverse the cribriform plate (CP) to reach the brain, we predicted that taxa with larger ORG repertoires would have proportionately expanded footprints in the CP foramina. Previous work found a correlation between ORG number and absolute CP size that disappeared after accounting for body size. Using updated, digital measurement data from high-resolution CT scans and re-examining the relationship between CP and body size, we report a striking linear correlation between relative CP area and number of functional ORGs across species from all mammalian superorders. This correlation suggests strong developmental links in the olfactory pathway between genes, neurons and skull morphology. Furthermore, because ORG number is linked to olfactory discriminatory function, this correlation supports relative CP size as a viable metric for inferring olfactory capacity across modern and extinct species. By quantifying CP area from a fossil sabertooth cat (Smilodon fatalis), we predicted a likely ORG repertoire for this extinct felid.


Type 1 Diabetes Treatments Based on Stem Cells.
More than a decade ago, a new research field named Stem Cell Therapy emerged in Health Science. Initially, it was considered that cells owned a differentiation capability; however, this dogma has changed when new results have been published regarding the ability of the cells to differentiate into different cell tissue mainly due to the novel reprogramming strategies. Accordingly, cells from an adult tissue source may be potentially capable of originating cells of a very different cell type. The possibility of transplanting these cells into damaged organs has triggered many studies to understand the plasticity of stem cells. Today, we have a deeper knowledge about stem cells, however still many questions, especially about the mechanism of action, that needs to be answered. The benefit of stem cells after transplantation has been demonstrated experimentally and also in some cases clinically; however, the extent of stem cell contribution in transplanted tissue has been found to be low and a large number of evidence indicates that a trophic effect should play an important role in such benefit. A better understanding of the paracrine mechanisms involved in this process could be of great relevance in order to focus studies on endogenous cells to direct their function towards the regeneration of damaged tissue. In addition, even more sophisticated methods of reprogramming and cell transplantation have been initiated in combination with bioengineering techniques in order to enhance the potential of these cells. In the present review, we will overview the studies on stem cell and their effects in the treatment of diabetes in order to discuss the questions generated about their origin and the mechanisms that are involved in their reparative properties.


Lycium barbarum extracts protect the brain from blood-brain barrier disruption and cerebral edema in experimental stroke.
Ischemic stroke is a destructive cerebrovascular disease and a leading cause of death. Yet, no ideal neuroprotective agents are available, leaving prevention an attractive alternative. The extracts from the fruits of Lycium barbarum (LBP), a Chinese anti-aging medicine and food supplement, showed neuroprotective function in the retina when given prophylactically. We aim to evaluate the protective effects of LBP pre-treatment in an experimental stroke model. C57BL/6N male mice were first fed with either vehicle (PBS) or LBP (1 or 10 mg/kg) daily for 7 days. Mice were then subjected to 2-hour transient middle cerebral artery occlusion (MCAO) by the intraluminal method followed by 22-hour reperfusion upon filament removal. Mice were evaluated for neurological deficits just before sacrifice. Brains were harvested for infarct size estimation, water content measurement, immunohistochemical analysis, and Western blot experiments. Evans blue (EB) extravasation was determined to assess blood-brain barrier (BBB) disruption after MCAO. LBP pre-treatment significantly improved neurological deficits as well as decreased infarct size, hemispheric swelling, and water content. Fewer apoptotic cells were identified in LBP-treated brains by TUNEL assay. Reduced EB extravasation, fewer IgG-leaky vessels, and up-regulation of occludin expression were also observed in LBP-treated brains. Moreover, immunoreactivity for aquaporin-4 and glial fibrillary acidic protein were significantly decreased in LBP-treated brains. Seven-day oral LBP pre-treatment effectively improved neurological deficits, decreased infarct size and cerebral edema as well as protected the brain from BBB disruption, aquaporin-4 up-regulation, and glial activation. The present study suggests that LBP may be used as a prophylactic neuroprotectant in patients at high risk for ischemic stroke.


RHD positive haplotypes in D negative Europeans.
Blood group genotyping is increasingly utilized for prenatal diagnosis and after recent transfusions, but still lacks the specificity of serology. In whites, the presence of antigen D is predicted, if two or more properly selected RHD-specific polymorphism are detected. This prediction must fail, if an antigen D negative RHD positive allele is encountered. Excluding RHDpsi and CdeS frequent only in individuals of African descent, most of these alleles are unknown and the population frequency of any such allele has not been determined. We screened 8,442 antigen D negative blood donations by RHD PCR-SSP. RHD PCR positive samples were further characterized by RHD exon specific PCR-SSP or sequencing. The phenotype of the identified alleles was checked and their frequencies in Germans were determined. We detected 50 RHD positive samples. Fifteen samples harbored one of three new Del alleles. Thirty samples were due to 14 different D negative alleles, only 5 of which were previously known. Nine of the 14 alleles may have been generated by gene conversion in cis, for which we proposed a mechanism triggered by hairpin formation of chromosomal DNA. The cumulative population frequency of the 14 D negative alleles was 1:1,500. Five samples represented a D+/- chimera, a weak D and three partial D, which had been missed by routine serology; two recipients transfused with blood of the D+/- chimera donor became anti-D immunized. The results of this study allowed to devise an improved RHD genotyping strategy, the false-positive rate of which was lower than 1:10,000. The number of characterized RHD positive antigen D negative and Del alleles was more than doubled and their population frequencies in Europe were defined.


Self-disorder and brain processing of proprioception in schizophrenia spectrum patients: a re-analysis.
Anomalies of self-awareness (self-disorders, SDs) are theorized to be basic to schizophrenia psychopathology. We have previously observed dysfunction of brain processing of proprioception in schizophrenia spectrum disorders (SZS). We hypothesized that SDs could be associated with abnormalities of early contralateral proprioceptive evoked oscillatory brain activity. We investigated the association between proprioceptive evoked potential components and SDs in a re-analysis of data from a subsample (n = 12) of SZS patients who had previously been observed with deviant proprioceptive evoked potentials and interviewed with the Examination of Anomalous Self-Experience (EASE) scale. Higher EASE scores (i.e. increased SD) were associated with lower peak parietal gamma frequencies and higher peak beta amplitudes over frontal and parietal electrodes in the left hemisphere following right-hand proprioceptive stimulation. Disorders of self-awareness may be associated with dysfunction of early phases of somatosensory processing. The findings are potentially relevant to our understanding of the pathophysiology of schizophrenia, but further studies are needed.


Comparison of Trabeculectomy Outcomes Performed by Residents in Training and Ophthalmologists.
Resident-performed trabeculectomies present higher intraocular pressure and lower complete success rate at 1 year. To compare the 1-year outcomes of ab externo trabeculectomy between residents in training and staff ophthalmologists. This retrospective study included all consecutive eyes submitted to ab externo trabeculectomy between January 2015 to June 2020. A 1-year complete success rate was considered using all the following criteria: intraocular pressure (IOP)≤21 mm Hg and ≥6mm Hg without ocular hypotensive medications; IOP reduction≥30%; without loss of light perception, phthisis bulbi, and further glaucoma surgery (excluding suture lysis and bleb needling). One hundred and ten eyes from 99 patients were included. Thirty percent (n=33) of the trabeculectomies were performed by residents. There were no significant preoperative differences between groups, apart from age at surgery, which was higher in the residents' group (72.39±6.83 vs. 62.00±15.07 years, P<0.001), and visual field index (Humphrey Field Analyzer), which was lower in the ophthalmologists' group (51.81±34.74% vs. 32.04±33.83%, P=0.013). IOP at 1-, 3-, 6 months, and 1 year after surgery was significantly higher in the resident's group (P<0.05). Resident-performed trabeculectomies achieved a significantly lower complete success rate when compared with the ophthalmologists' group (39.39% vs. 64.94%, P=0.013). The overall rate of the postoperative complications and reintervention did not differ between groups, but the occurrence of a shallow anterior chamber was more frequent in the residents' group (15.15% vs. 4.05%, P=0.037). Resident-performed trabeculectomies present significantly higher postoperative IOP levels and a lower complete success rate when compared with staff ophthalmologists. It is, therefore, fundamental to adopt strategies to change this gap, improve patient safety, and strengthen resident confidence.


Mathematical modeling of aeration efficiency and dissolved oxygen provided by stepped cascade aeration.
Mathematical modeling has been a vital tool in the field of environmental engineering. Various models have been developed to simulate the level of aeration efficiency (AE) provided by different aerating structures to raise levels of dissolved oxygen (DO) in streams; one of which is the stepped cascade structure. Three models developed by Gameson et al. WRL, and Nakasone, in addition to Qual2k, a computer program for stream modeling, have been used in this research; values of AEs obtained have been compared to those computed using DO measured from a built model at a WWTP. A stepped cascade structure was installed with different heights to aerate five flowrates with different levels of COD. An adjustment has been made to the Nakasone model to test the effect of pollutant load on the amount of aeration that could be reached. Values of AEs computed using the Gameson model were 30%, 39.5%, and 40% for cascade heights (H(d)) 45, 60, and 75 cm respectively for the five flowrates (q) that ranged from 21-66 m³/hr. Values of AEs from WRL model were 32.8%, 42%, and 43% consequently. Values of AEs from Nakasone model ranged from 4.6-7.5%, 6-10%, and 7.6-12% respectively. For the adjusted Nakasone model, values of AEs ranged from 3.2-4.9%, 3.3-5.3%, and 4.1-6.7% respectively. Finally, the AEs computed using the values of downstream DO generated by Qual2k ranged from 4-18%, 2-15%, and 2.5-5.1% correspondingly. Around 80% of the downstream DO values computed using the Nakasone and adjusted Nakasone model were closer to those measured in the field, thus more reliable in cascade design.


Two homeopathic remedies used intermittently provide additional protective effects against hepatotoxicity induced by carcinogens in mice.
The purpose of the study was to evaluate whether potentized cholesterinum (Chol) intermittently used with another homeopathic remedy, Natrum Sulphuricum (Nat Sulph) can provide additional benefits in combating hepatotoxicity generated by chronic feeding of carcinogens, p-dimethylaminoazobenzene (p-DAB), and phenobarbital (PB). Mice were categorized into subgroups: normal untreated (Gr-1); normal + alcohol "vehicle" (Alc) (Gr-2), 0.06% p-DAB +0.05% PB (Gr-3), p-DAB+PB+Alc (Gr-4), p-DAB+PB+Nat Sulph-30 (Gr-5), p-DAB+PB+Chol-200 (Gr-6), p-DAB+PB+Nat Sulph-30+Chol-200 (Gr-7), p-DAB+PB+Nat Sulph-200 (Gr-8), and DAB+PB+Nat Sulph-200+Chol-200 (Gr-9). Hepatotoxicity was assessed through biomarkers like aspartate and alanine aminotransferases (AST and ALT), acid and alkaline phosphatases (AcP and AlkP), reduced glutathione content (GSH), glucose 6-phosphate dehydrogenase (G6PD), gamma glutamyl transferase (GGT), lactate dehydrogenase (LDH), and analysis of lipid peroxidation (LPO) at 30, 60, 90, and 120 days and antioxidant biomarkers like superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) were assayed. Electron microscopic studies (scanning and transmission) and gelatin zymography for matrix metalloproteinases were conducted in liver. The feeding of the homeopathic drugs showed intervention in regard to the increased activities of AST, ALT, AcP, AlkP, GGT, LDH, and LPO and decreased activities of G6PD, SOD, CAT, GR, and GSH noted in the intoxicated mice, more appreciable in Groups 7 and 9. Thus, combined therapy provided additional antihepatotoxic and anticancer effects.


Immune cell membrane fatty acids and inflammatory marker, C-reactive protein, in patients with multiple sclerosis.
Measurement of fatty acids in biological fluids and cell membranes including leucocytes from multiple sclerosis patients is inconsistent. The objective of the present study was to investigate the fatty acid composition within the different membrane phospholipid fractions in peripheral blood mononuclear cells in multiple sclerosis patients, and correlate with severity of neurological outcome as measured by the Kurtzke Expanded Disability Status Scale and Functional System Scores. The fatty acid composition of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, sphingomyelin and phosphatidylinositol phospholipids in the peripheral blood mononuclear cells of twenty-six multiple sclerosis and twenty-five control subjects were measured by GC, and C-reactive protein was measured in all subjects. The elongation product of 20 : 4n-6, 22 : 4n-6, was significantly decreased in membrane phosphatidylethanolamine and phosphatidylserine in multiple sclerosis patients (P = 0.01 and P = 0.03 respectively), and correlated inversely with severity of disease and C-reactive protein. Also an inverse correlation was observed between the C-reactive protein and membrane phosphatidylcholine and phosphatidylserine 20 : 4n-6. Cultural and ethnic differences, as well as dietary variability, especially in a diseased state have been implicated in the differences observed in the fatty acid composition in peripheral blood mononuclear cell membranes of patients with multiple sclerosis. The present results suggest that the disease state may in part explain the reported inconsistencies in fatty acid levels in multiple sclerosis patients.


The role and limits of a gradient based explanation of morphogenesis: a theoretical consideration.
Development of an organism is a multi-dimensional process leading to the generation of complex species-specific structures. This specificity suggests machine-like organisation. The uneven distribution (gradient) of soluble substances (morphogens) and specific receptor-ligand interactions are known to cause differential gene expression. Therefore gradients of morphogens are used as a causal explanation of developmental processes. However each attempt to describe development causally should take into account both the local fine organisation and global robustness of morphogenesis. The classical view of the role of morphogens will be critically considered and possible alternative proposed. The core idea of my proposal is that the main function of morphogenetic substances could be a context dependent modification of cell behaviour. Both history and different features of morphogenetic fields create the framework for the activity of morphogenes.


Methods for pre-testing and piloting survey questions: illustrations from the KENQOL survey of health-related quality of life.
Part of the assertion that any survey researcher can make about the validity of their results needs to contain an analysis of questions and their responses from the respondent's viewpoint. Claims concerning the validity, reliability and sensitivity of health-related quality of life measures tend to be based on the quantitative approach of psychometrics, which fails to identify when respondents: misinterpret questions; do not recall the information requested; or give answers that present themselves in a better or worse light. The paper presents some approaches to pre-testing and piloting survey questionnaires to check the interpretation of survey questions, using illustrations from the KENQOL project. The paper describes: how the intended referential and connotative meaning of each question was established; the criteria to judge the appropriateness of each question; the methods used to make those judgements; and the process of reviewing questions based on findings. The role of piloting is highlighted, and further reading is suggested for readers wishing to develop a model for their own investigation.


Functional expression of cell surface cannabinoid CB(1) receptors on presynaptic inhibitory terminals in cultured rat hippocampal neurons.
At present, little is known about the mechanisms by which cannabinoids exert their effects on the central nervous system. In this study, fluorescence imaging and electrophysiological techniques were used to investigate the functional relationship between cell surface cannabinoid type 1 (CB(1)) receptors and GABAergic synaptic transmission in cultured hippocampal neurons. CB(1) receptors were labelled on living neurons using a polyclonal antibody directed against the N-terminal 77 amino acid residues of the rat cloned CB(1) receptor. Highly punctate CB(1) receptor labelling was observed on fine axons and at axonal growth cones, with little somatic labelling. The majority of these sites were associated with synaptic terminals, identified either with immunohistochemical markers or by using the styryl dye FM1-43 to label synaptic vesicles that had undergone active turnover. Dual labelling of neurons for CB(1) receptors with either the inhibitory neurotransmitter GABA or its synthesising enzyme glutamate decarboxylase, demonstrated a strong correspondence. The immunocytochemical data was supported by functional studies using whole-cell patch-clamp recordings of miniature inhibitory postsynaptic currents (mIPSCs). The cannabinoid agonist WIN55,212-2 (100nM) markedly inhibited (by 77+/-6.3%) the frequency of pharmacologically-isolated GABAergic mIPSCs. The effects of WIN55,212-2 were blocked in the presence of the selective CB(1) receptor antagonist SR141716A (100nM).In conclusion, the present data show that cell surface CB(1) receptors are expressed at presynaptic GABAergic terminals, where their activation inhibits GABA release. Their presence on growth cones could indicate a role in the targeting of inhibitory connections during development.


The Impact of Insurance Status on Tumor Characteristics and Treatment Selection in Contemporary Patients With Prostate Cancer.
Approximately 15% of the US population does not have health insurance. The objective of this study was to evaluate the impact of insurance status on tumor characteristics and treatment selection in patients with prostate cancer. We identified 20,393 patients younger than 65 years with prostate cancer in the 2010-2011 SEER database. Multivariable logistic regression analysis tested the relationship between insurance status and 2 end points: (1) presenting with low-risk prostate cancer at diagnosis and (2) receiving local treatment of the prostate. Locally weighted scatterplot smoothing methods were used to graphically explore the interaction among insurance status, use of local treatment, and baseline risk of cancer recurrence. The latter was defined using the Stephenson nomogram and CAPRA score. Overall, 18,993 patients (93%) were insured, 849 (4.2%) had Medicaid coverage, and 551 (2.7%) were uninsured. At multivariable analysis, Medicaid coverage (odds ratio [OR], 0.67; 95% CI, 0.57, 0.80; P<.0001) and uninsured status (OR, 0.57; 95% CI, 0.46, 0.71; P<.0001) were independent predictors of a lower probability of presenting with low-risk disease. Likewise, Medicaid coverage (OR, 0.72; 95% CI, 0.60, 0.86; P=.0003) and uninsured status (OR, 0.45; 95% CI, 0.37, 0.55; P<.0001) were independent predictors of a lower probability of receiving local treatment. In uninsured patients, treatment disparities became more pronounced as the baseline cancer recurrence risk increased (10% in low-risk patients vs 20% in high-risk patients). Medicaid beneficiaries and uninsured patients are diagnosed with higher-risk disease and are undertreated. The latter is more accentuated for patients with high-risk prostate cancer. This may seriously compromise the survival of these individuals.


Antigens induced on erythrocytes by P. falciparum: expression of diverse and conserved determinants.
Red blood cells that are infected with the malaria parasite Plasmodium falciparum express new antigens on their surface. In a study of these antigens in the erythrocytes of naturally infected children in the Gambia, an antibody-mediated agglutination assay revealed an extreme degree of antigenic diversity. Serum samples from each of ten children in the convalescent stage of malaria infection reacted with infected cells from the same child but generally not with infected cells from the other children. The Gambian children's erythrocytes also expressed shared determinants: sera from Gambian adults often reacted with the surface of infected cells from all of the children and were shown by adsorption and elution experiments to contain antibodies that recognized several isolates. Conserved determinants exposed on infected erythrocytes may be important for development of antimalarial immunity either naturally or through vaccination.


Human aggrecan keratan sulfate undergoes structural changes during adolescent development.
Alkaline borohydride-reduced keratan sulfate chains were isolated from human articular cartilage aggrecan from individuals of various ages (0-85 years old). The chains were structurally characterized using 1H NMR spectroscopy, gel permeation chromatography, and oligosaccharide profiling (after digestion with the enzymes keratanase and keratanase II). The results show that from birth to early adolescence (0-9 years) the levels of alpha(1-3)-fucosylation, alpha(2-3)-sialylation, and galactose sulfation increase. Also, the weight-average molecular weight of the chains increases. During maturation (9-18 years) the levels of fucosylation and galactose sulfation continue to increase and alpha(2-6)-sialylation of the chains occurs. In adult life (18-85 years) there is little change in the weight-average molecular weight of the chains, and the levels of fucosylation, sialylation, and sulfation remain fairly constant.


Customizing Structure-Function Displacements in the Macula for Individual Differences.
In the macula, retinal ganglion cells (RGCs) are displaced from their receptive fields. We used optical coherence tomography (OCT) to customize displacements for individual eyes by taking into account macular shape parameters, and determined the likely effect of individual anatomical differences on structure-function mapping in the central visual field. Using the population average model of Drasdo et al. as a starting point, we altered the RGC count in that model based on the ratio of an individual's RGC layer plus inner plexiform layer thickness to the population average on a pointwise basis as a function of eccentricity from the fovea. For 20 adults (age, 24-33; median age, 28) with normal vision, we computed displacements with the original model and our customized approach. We report the variance in displacements among individuals and compare the effects of such displacements on structure-function mapping of the commonly used the 10-2 visual field pattern. As expected, customizing the displacement using individual OCT data made only a small difference on average from the population-based values predicted by the Drasdo et al. model. However, the range between individuals was over 1° at many locations, and closer to 2° at some locations in the superior visual field. Individualizing macular displacement measurements based on OCT data for an individual can result in large spatial shifts in the retinal area corresponding to 10-2 locations, which may be important for clinical structure-function analysis when performed on a local, spatial scale.


Organic semiconductor distributed feedback laser pixels for lab-on-a-chip applications fabricated by laser-assisted replication.
The integration of organic semiconductor distributed feedback (DFB) laser sources into all-polymer chips is promising for biomedical or chemical analysis. However, the fabrication of DFB corrugations is often expensive and time-consuming. Here, we apply the method of laser-assisted replication using a near-infrared diode laser beam to efficiently fabricate inexpensive poly(methyl methacrylate) (PMMA) chips with spatially localized organic DFB laser pixels. This time-saving fabrication process enables a pre-defined positioning of nanoscale corrugations on the chip and a simultaneous generation of nanoscale gratings for organic edge-emitting laser pixels next to microscale waveguide structures. A single chip of size 30 mm × 30 mm can be processed within 5 min. Laser-assisted replication allows for the subsequent addition of further nanostructures without a negative impact on the existing photonic components. The minimum replication area can be defined as being as small as the diode laser beam focus spot size. To complete the fabrication process, we encapsulate the chip in PMMA using laser transmission welding.


Large punctuational contribution of speciation to evolutionary divergence at the molecular level.
A long-standing debate in evolutionary biology concerns whether species diverge gradually through time or by punctuational episodes at the time of speciation. We found that approximately 22% of substitutional changes at the DNA level can be attributed to punctuational evolution, and the remainder accumulates from background gradual divergence. Punctuational effects occur at more than twice the rate in plants and fungi than in animals, but the proportion of total divergence attributable to punctuational change does not vary among these groups. Punctuational changes cause departures from a clock-like tempo of evolution, suggesting that they should be accounted for in deriving dates from phylogenies. Punctuational episodes of evolution may play a larger role in promoting evolutionary divergence than has previously been appreciated.


A score for estimating survival in idiopathic pulmonary fibrosis with rest SpO2>88.
Prediction models for survival at baseline evaluation have been proposed in IPF but some are difficult to use. The aim of present study was to develop a new mortality risk scoring system for patients with IPF at initial evaluation. One hundred twenty with definitive IPF were selected through a review of standardized medical records for interstitial lung diseases. Patients with resting SpO2<89% were excluded. Significant individual predictors we derived by a Cox proportional hazards model and transformed in categorical data according to cut-off points. Beta coefficients for each predictor were similar, so a score was created considering the sum of dichotomic (0 or 1) transformed variables. Median follow-up time was 37.5 months. At the end of follow-up, 80 patients had died. Independent predictors of mortality by multivariate analysis included dyspnea (at rest or to light or moderate activities), FVC<70%,FEV1/FVC>0.89 and DLCO ≤ 40%. Resting SpO2 and ExSpO2 were excluded in final analysis. The hazard ratios ranged from 1.95 for dyspnoea to 2.30 for DLCO. When the total score was 0 (Stage I, n=28), median survival time was 68 months; when 1 or 2 (Stage II, n=69), it was 45 months; and when 3 or 4 (Stage III, n=23), it was 19 months (log rank= 60.44, p<0.001). The score can separate IPF patients with high, intermediate and low survival.


Inhibitory effects of zingerone, a pungent component of Zingiber officinale Roscoe, on colonic motility in rats.
Ginger (rhizome of Zingiber officinale Roscoe) is an herbal medicine for the treatment of gastrointestinal disorders including constipation and diarrhea. Zingerone is a likely active constituent responsible for the antidiarrheal activity of ginger. The current study was designed to characterize pharmacological actions of zingerone on colonic motility. To evaluate pharmacological effects of zingerone on colonic motility, we used isolated colonic segments from rats, in which mechanical responses were recorded in the longitudinal direction. In addition, we evaluated the effects on colonic motility in vivo by measuring intraluminal pressure changes and expelled fluid volume from the colon in anesthetized rats. Zingerone was applied to the lumen of the colon to allow the drug to access from the mucosal side. Zingerone inhibited spontaneous contractile movements in the isolated colonic segments in a dose-dependent manner. The inhibitory effects of zingerone on colonic movements were not affected by pretreatment with capsazepine, a typical antagonist of transient receptor potential vanilloid 1. In addition, tetrodotoxin, a blocker of voltage-dependent sodium channels on neurons, did not affect the suppression of colonic movements by zingerone, suggesting that zingerone acts on the smooth muscles directly. Zingerone also attenuated colonic motility in vivo without affecting blood pressure and heart rate. The effects were reversible and reproducible. Our findings suggest that zingerone can inhibit colonic motility via direct action on smooth muscles. Zingerone might exert beneficial therapeutic effects on hypermotility-induced diarrhea by abrogating excessive gastrointestinal motility.


Establishing New Community-Based Participatory Research Partnerships using the Community-Based Participatory Research Charrette Model: Lessons from the Cancer Health Accountability for Managing Pain and Symptoms Study.
Community-based participatory research (CBPR) is a collaborative and equitable approach to research inquiry; however, the process of establishing and maintaining CBPR partnerships can be challenging. There is an ongoing need for innovative strategies that foster partnership development and long-term sustainability. In 2010, the University of North Carolina at Chapel Hill developed a CBPR charrette model to facilitate stakeholder engagement in translational research. To describe how the Cancer Health Accountability for Managing Pain and Symptoms (CHAMPS) Study leveraged the CBPR charrette process to develop and strengthen its CBPR partnership and successfully implement research objectives. Fourteen CHAMPS community, academic, and medical partners participated in the CBPR charrette. Two co-facilitators guided the charrette application process and in-person discussion of partnership strengths, needs, and challenges. Community experts (CEs) and academic experts (AEs) with extensive experience in CBPR and health disparities provided technical assistance and recommendations during the in-person charrette. Overall, the CHAMPS partnership benefited significantly from the charrette process. Specifically, the charrette process engendered greater transparency, accountability, and trust among CHAMPS partners by encouraging collective negotiation of project goals and implementation, roles and responsibilities, and compensation and communication structures. The process also allowed for the exploration of newly identified challenges and potential solutions with support from CEs and AEs. Furthermore, the charrette also functioned as a catalyst for capacity building among CHAMPS community, academic, and medical partners. Future studies should compare the impact of the CBPR charrette, relative to other approaches, on partnership development and process evaluation outcomes.


Efficacy methods to evaluate health communication and marketing campaigns.
Communication and marketing are growing areas of health research, but relatively few rigorous efficacy studies have been conducted in these fields. In this article, we review recent health communication and marketing efficacy research, present two case studies that illustrate some of the considerations in making efficacy design choices, and advocate for greater emphasis on rigorous health communication and marketing efficacy research and the development of a research agenda. Much of the outcomes research in health communication and marketing, especially mass media, utilizes effectiveness designs conducted in real time, in the media markets or communities in which messages are delivered. Such evaluations may be impractical or impossible, however, imiting opportunities to advance the state of health communication and marketing research and the knowledge base on effective campaign strategies, messages, and channels. Efficacy and effectiveness studies use similar measures of behavior change. Efficacy studies, however, offer greater opportunities for experimental control, message exposure, and testing of health communication and marketing theory. By examining the literature and two in-depth case studies, we identify advantages and limitations to efficacy studies. We also identify considerations for when to adopt efficacy and effectiveness methods, alone or in combination. Finally, we outline a research agenda to investigate issues of internal and external validity, mode of message presentation, differences between marketing and message strategies, and behavioral outcomes.


Increased transcript levels and kinetic function of pyruvate kinase during severe dehydration in aestivating African clawed frogs, Xenopus laevis.
The African clawed frog, Xenopus laevis, can withstand extremely arid conditions through aestivation, resulting in dehydration and urea accumulation. Aestivating X. laevis reduce their metabolic rate, and rely on anaerobic glycolysis to meet reduced ATP demands. The present study investigated how severe dehydration affected the transcript levels, kinetic profile, and phosphorylation state of the key glycolytic enzyme pyruvate kinase (PK) in the liver and skeletal muscle of X. laevis. Compared to control frogs, severely dehydrated frogs showed an increase in the transcript abundance of both liver and muscle isoforms of PK. While the kinetics of muscle PK did not differ between dehydrated and control frogs, PK from the liver of dehydrated frogs had a lower Km for phosphoenolpyruvate (PEP) (38%), a lower Ka for fructose-1,6-bisphosphate (F1,6P2) (32%), and a greater activation of PK via F1,6P2 (1.56-fold). PK from dehydrated frogs also had a lower phosphorylation-state (25%) in comparison to the enzyme from control frogs in the liver. Experimental manipulation of the phosphorylation-state of liver PK taken from control frogs by endogenous protein phosphatases resulted in decreased phosphorylation, and a similar kinetic profile as seen in dehydrated frogs. The physiological consequence of dehydration-induced PK modification appears to adjust PK function to remain active during a metabolically depressed state. This study provides evidence for the maintenance of PK activity through elevated mRNA levels and a dephosphorylation event which activates frog liver PK in the dehydrated state in order to facilitate the production of ATP via anaerobic glycolysis.


Tocilizumab in Patients Hospitalized with Covid-19 Pneumonia.
Coronavirus disease 2019 (Covid-19) pneumonia is often associated with hyperinflammation. Despite the disproportionate incidence of Covid-19 among underserved and racial and ethnic minority populations, the safety and efficacy of the anti-interleukin-6 receptor antibody tocilizumab in patients from these populations who are hospitalized with Covid-19 pneumonia are unclear. We randomly assigned (in a 2:1 ratio) patients hospitalized with Covid-19 pneumonia who were not receiving mechanical ventilation to receive standard care plus one or two doses of either tocilizumab (8 mg per kilogram of body weight intravenously) or placebo. Site selection was focused on the inclusion of sites enrolling high-risk and minority populations. The primary outcome was mechanical ventilation or death by day 28. A total of 389 patients underwent randomization, and the modified intention-to-treat population included 249 patients in the tocilizumab group and 128 patients in the placebo group; 56.0% were Hispanic or Latino, 14.9% were Black, 12.7% were American Indian or Alaska Native, 12.7% were non-Hispanic White, and 3.7% were of other or unknown race or ethnic group. The cumulative percentage of patients who had received mechanical ventilation or who had died by day 28 was 12.0% (95% confidence interval [CI], 8.5 to 16.9) in the tocilizumab group and 19.3% (95% CI, 13.3 to 27.4) in the placebo group (hazard ratio for mechanical ventilation or death, 0.56; 95% CI, 0.33 to 0.97; P = 0.04 by the log-rank test). Clinical failure as assessed in a time-to-event analysis favored tocilizumab over placebo (hazard ratio, 0.55; 95% CI, 0.33 to 0.93). Death from any cause by day 28 occurred in 10.4% of the patients in the tocilizumab group and 8.6% of those in the placebo group (weighted difference, 2.0 percentage points; 95% CI, -5.2 to 7.8). In the safety population, serious adverse events occurred in 38 of 250 patients (15.2%) in the tocilizumab group and 25 of 127 patients (19.7%) in the placebo group. In hospitalized patients with Covid-19 pneumonia who were not receiving mechanical ventilation, tocilizumab reduced the likelihood of progression to the composite outcome of mechanical ventilation or death, but it did not improve survival. No new safety signals were identified. (Funded by Genentech; EMPACTA ClinicalTrials.gov number, NCT04372186.).


Efficacy of leupeptin, superoxide dismutase, and verapamil in modulating delayed reperfusion damage after burn injury.
Reperfusion damage has been identified as an important factor in multiorgan failure after severe burn injury. We wondered if leupeptin, a protease inhibitor, superoxide dismutase (SOD), a scavenger of free oxygen radicals, or verapamil, a calcium antagonist, would protect the cellular energy metabolism when they were given with fluid resuscitation that was delayed 6 hours after a severe burn injury. Fifty male rats weighing 280 to 300 gm received a 50% third-degree scald burn. Ten of these received fluid resuscitation at 30 minutes and 1 1/2 hours after injury, and 40 received delayed fluid resuscitation at 6 and 7 hours after injury. Thirty of these 40 rats were given leupeptin (n = 10), SOD (n = 10), or verapamil (n = 10). Heart, liver, and kidney tissue samples were obtained 8 hours after injury; adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate were measured; and the energy charge potential was calculated. Tissue water content (TWC) in lung and skeletal muscle was also determined. The adenine nucleotide pool and the energy charge potential in heart, liver, and kidney tissue were all significantly decreased (p < 0.01) in rats receiving delayed fluid resuscitation compared with those receiving early resuscitation. Leupeptin was effective in protecting the heart against reperfusion damage, and verapamil and leupeptin showed some efficacy in protecting kidney tissue. Liver tissue, however, showed no protective response with therapy. TWC was significantly decreased (p < 0.01) in skeletal muscle with SOD treatment, and though all treatments appeared to keep lung water content reduced, none was significant at p < 0.01. We thus conclude that both the decreases in heart and kidney adenine nucleotides and the increase in TWC that are caused by delayed fluid resuscitation can be attenuated with appropriate pharmacologic agents.


Mobile suitcase laboratory for rapid detection of Leishmania donovani using recombinase polymerase amplification assay.
Leishmania donovani (LD) is a protozoan parasite transmitted to humans from sand flies, which causes Visceral Leishmaniasis (VL). Currently, the diagnosis is based on presence of the anti-LD antibodies and clinical symptoms. Molecular diagnosis would require real-time PCR, which is not easy to implement at field settings. In this study, we report on the development and testing of a recombinase polymerase amplification (RPA) assay for the detection of LD. A genomic DNA sample was applied to determine the assay analytical sensitivity. The cross-reactivity of the assay was tested by DNA of Leishmania spp. and of pathogens considered for differential diagnosis. The clinical performance of the assay was evaluated on LD positive and negative samples. All results were compared with real-time PCR. To allow the use of the assay at field settings, a mobile suitcase laboratory (56 × 45.5 × 26.5 cm) was developed and operated at the local hospital in Mymensingh, Bangladesh. The LD RPA assay detected equivalent to one LD genomic DNA. The assay was performed at constant temperature (42 °C) in 15 min. The RPA assay also detected other Leishmania species (L. major, L. aethiopica and L. infantum), but did not identify nucleic acid of other pathogens. Forty-eight samples from VL, asymptomatic and post-kala-azar dermal leishmaniasis subjects were detected positive and 48 LD-negative samples were negative by both LD RPA and real-time PCR assays, which indicates 100 % agreement. The suitcase laboratory was successfully operated at the local hospital by using a solar-powered battery. DNA extraction was performed by a novel magnetic bead based method (SpeedXtract), in which a simple fast lysis protocol was applied. Moreover, All reagents were cold-chain independent. The mobile suitcase laboratory using RPA is ideal for rapid sensitive and specific detection of LD especially at low resource settings and could contribute to VL control and elimination programmes.


Receptor units responding to movement in the octopus mantle.
1. A preparation of the mantle of Octopus which is inverted over a solid support and which exposes the stellate ganglion and associated nerves is described. 2. Afferent activity can be recorded from stellar nerves following electrical stimulation of the pallial nerve. The latency and frequency of the phasic sensory response is correlated with the contraction of the mantle musculature. 3. It is proposed that receptors cells located in the muscle, and their activity following mantle contraction, form part of a sensory feedback system in the mantle. Large, multipolar nerve cells that were found between the two main layers of circular muscle in the mantle could be such receptors.


A comprehensive three-dimensional cortical map of vowel space.
Mammalian cortex is known to contain various kinds of spatial encoding schemes for sensory information including retinotopic, somatosensory, and tonotopic maps. Tonotopic maps are especially interesting for human speech sound processing because they encode linguistically salient acoustic properties. In this study, we mapped the entire vowel space of a language (Turkish) onto cortical locations by using the magnetic N1 (M100), an auditory-evoked component that peaks approximately 100 msec after auditory stimulus onset. We found that dipole locations could be structured into two distinct maps, one for vowels produced with the tongue positioned toward the front of the mouth (front vowels) and one for vowels produced in the back of the mouth (back vowels). Furthermore, we found spatial gradients in lateral-medial, anterior-posterior, and inferior-superior dimensions that encoded the phonetic, categorical distinctions between all the vowels of Turkish. Statistical model comparisons of the dipole locations suggest that the spatial encoding scheme is not entirely based on acoustic bottom-up information but crucially involves featural-phonetic top-down modulation. Thus, multiple areas of excitation along the unidimensional basilar membrane are mapped into higher dimensional representations in auditory cortex.


Effects of bisphenol A on decreasing the percentage and promoting the growth of stem cell-like cells from SK-N-SH human neuroblastoma cells.
Bisphenol A (BPA) is an industrial contaminant and is reported to be a risk factor associated with the development of tumors. In our previous studies, we have shown that BPA promoted the growth of SK-N-SH human neuroblastoma cells and increased their invasion and metastasis. In this study, we further investigated the effects of BPA and 17β-estradiol (E2) on the stem cell-like cells from SK-N-SH cells. Detection of stem cell markers, proliferation assay, and clonogenic analysis showed that the side-population (SP) of SK-N-SH cells had properties similar to those of stem cells. BPA or E2 exposure decreased the percentage of SP cells and the expression of stem cell-marker proteins. BPA and E2 promoted the growth of non-SP cells to a greater extent than of SP cells; in addition, they significantly increased the growth of SP cells. Thus, BPA has effects on stem cell-like cells, which induce tumor formation, and thus, BPA is an environmental factor that plays an important role in the development of neuroblastoma.


Hydroxylated-graphene quantum dots induce cells senescence in both p53-dependent and -independent manner.
The numerous particular chemical/physical properties make graphene quantum dots (GQDs) attractive for various biomedical applications such as drug delivery, bioimaging and tumor photodynamic therapy (PDT). In the present study, the critical roles of hydroxyl-modified GQDs (OH-GQDs) on lung carcinoma A549 (wild type p53) and H1299 (p53-null) cells were investigated. Our data showed that a medium concentration (50 μg mL-1) of OH-GQDs significantly decreased the viability of A549 and H1299 cells. OH-GQDs treatment enhanced intracellular reactive oxygen species (ROS) generation. Furthermore, we found that treatment with ROS scavenger N-acetylcysteine (NAC) at least partially abolished the cytotoxic effect of OH-GQDs on A549 and H1299 cells. Hydroxylated GQDs lead to G0-G1 arrest and cells senescence. Signal pathway analysis revealed that OH-GQDs activated the expression of p21 in both a p53-dependent and -independent manner. Consistent with this, OH-GQDs could also inhibit the phosphorylation of Rb in both A549 and H1299 cells. These findings provide valuable information for the consideration of biomedical application of GQDs in the future.


Mental Health and Functioning of Female Sex Workers in Chittagong, Bangladesh.
To examine the mental health and functioning of female sex workers (FSW) in Chittagong, Bangladesh, a population that has commonly been neglected in mental health policy and research. We included 259 women in the study (M age: 23.2 years; range: 11-48). The comprehensive Composite International Diagnostic Interview was used to assess their 12-month prevalence rates of DSM-IV mental disorders, and a comprehensive questionnaire was adapted to explore various factors, such as socio-demographics, working and living conditions, or experiences of abuse. On average, participants began their commercial sex work at 18.5 years old (range: 10-45). Their main motives for sex work were coercion (49.4%) and the necessity to financially support families (54.8%). In total, 224 FSW (86.5%) wanted to quit commercial sex work. A mental disorder within the past 12 months was reported by 100 FSW (38.6%), with drug abuse clearly being the most prevalent diagnosis (15.4%). Sexual, physical, and emotional abuse were very common among the FSW, and substance use disorders (SUD) were significantly more prevalent in persons who experienced emotional abuse (OR = 2.2). Prevalence rates of any mental disorder and SUD were higher in women who did sex work to support their family, whereas mood disorders were more frequent in those who needed the money to pay debts. Participants with any disorder were significantly older than those without (M age: 24.4 vs. 22.5 years) and had started significantly later in the sex business (M age: 19.7 vs. 17.7 years). Our study revealed that FSW in Chittagong are very vulnerable and highly impaired, as expressed by high rates of abuse and mental disorders. Coercion is very common and many FSW are required to work in the sex business because they need the money to support their families. FSW are a very marginalized population, especially in the developing countries where awareness for mental health is low and the availability of mental health services is insufficient.


Appearances are deceptive...
A 31-year-old man was referred with sudden onset of a painful periorbital angio-oedema. Assuming a first episode of allergic or acquired angio-oedema, the patient was treated with a plasma-derived C1 esterase inhibitor, tranexamic acid and fresh-frozen plasma, after the initial treatment with corticosteroids and histamine blockers was unsuccessful. As blood cultures grew group A streptococci, the diagnosis of periorbital necrotising fasciitis was made, and treatment was subsequently changed to clindamycin and ceftriaxone. Necrotising fasciitis involving the periorbita is a rare and potentially lethal differential diagnosis of acute angio-oedema, in particular if accompanied by severe pain and/or fever. Knowledge on the appearance of this disorder cannot be overemphasised, as only early recognition and immediate initiation of high-dose penicillin and clindamycin combined with tissue debridement help to decrease the mortality and morbidity.


Usability evaluation of a GUI prototype for a ventilator machine.
Information presentation on the monitor screen of ventilator machines affects nurses' response and decision-making during ventilation treatment. The purpose of this study is to evaluate whether a new GUI (graphical user interface) prototype, the so-called circular display prototype can make deviations easy to detect. A numerical display prototype was made and used as a reference display in the evaluation. Six task scenarios that involved parameter changes were selected to simulate a real situation under volume control (VC) mode during ventilation treatment. Usability tests with the two display designs were carried out in a usability laboratory. Twenty medical nursing students participated as test subjects in the usability tests. The objective results showed that the graphical circular display had an advantage over the numerical display in interpreting parameter changes, but not in reducing the error rates for detecting the number of parameter changes or for forming an overall picture of the patient's situation. Furthermore, the circular display prototype did not improve the detection time. Although the majority of the test subjects preferred the graphical circular display, the results implied that several aspects of this prototype should be improved in a future development study.


[Health Effects of Asian Dust Events: A Literature Review Update of Epidemiological Evidence].
Asian dust is a phenomenon involving the long-range transport of atmospheric pollutants originating from the desert areas of China and Mongolia. In recent years, the health effects of Asian dust have raised public concerns. Numerous studies on the health effects of Asian dust have been published since the last review in 2010. Thus, a literature review was conducted to shed light on the latest epidemiologic findings. PubMed and Science Direct databases were used for the review of epidemiologic studies published between June 2009 and April 2018. We identified 53 epidemiologic studies. Mortality, ambulance transportation, hospitalization/medical examination, changes in symptomatic, functional, and examination findings, as well as birth outcomes have been reported as outcomes. When the outcomes were categorized by disease, the effects of Asian dust on respiratory, cardiovascular, and allergic diseases raised concerns. The common evidences of causation between Asian dust and these diseases were the consistency of findings and temporal sequence of association. As results of research on dose-response relationships have become available, and the possibility that the health effects of Asian dust may vary depending on its chemical composition has been pointed out, further research using the exposure level indicators of Asian dust or its chemical composition should be conducted. Furthermore, with focus on the crucial issue of reducing exposure, research related to prevention and raising awareness should be further promoted.


Spontaneous domain formation in disordered copolymers as a mechanism for chromosome structuring.
Motivated by the problem of domain formation in chromosomes, we studied a co-polymer model where only a subset of the monomers feel attractive interactions. These monomers are displaced randomly from a regularly-spaced pattern, thus introducing some quenched disorder in the system. Previous work has shown that in the case of regularly-spaced interacting monomers this chain can fold into structures characterized by multiple distinct domains of consecutive segments. In each domain, attractive interactions are balanced by the entropy cost of forming loops. We show by advanced replica-exchange simulations that adding disorder in the position of the interacting monomers further stabilizes these domains. The model suggests that the partitioning of the chain into well-defined domains of consecutive monomers is a spontaneous property of heteropolymers. In the case of chromosomes, evolution could have acted on the spacing of interacting monomers to modulate in a simple way the underlying domains for functional reasons.


Template-based CTA to x-ray angio rigid registration of coronary arteries in frequency domain with automatic x-ray segmentation.
A key challenge for image guided coronary interventions is accurate and absolutely robust image registration bringing together preinterventional information extracted from a three-dimensional (3D) patient scan and live interventional image information. In this paper, the authors present a novel scheme for 3D to two-dimensional (2D) rigid registration of coronary arteries extracted from preoperative image scan (3D) and a single segmented intraoperative x-ray angio frame in frequency and spatial domains for real-time angiography interventions by C-arm fluoroscopy. Most existing rigid registration approaches require a close initialization due to the abundance of local minima and high complexity of search algorithms. The authors' method eliminates this requirement by transforming the projections into translation-invariant Fourier domain for estimating the 3D pose. For 3D rotation recovery, template Digitally Reconstructed Radiographs (DRR) as candidate poses of 3D vessels of segmented computed tomography angiography are produced by rotating the camera (image intensifier) around the DICOM angle values with a specific range as in C-arm setup. The authors have compared the 3D poses of template DRRs with the segmented x-ray after equalizing the scales in three domains, namely, Fourier magnitude, Fourier phase, and Fourier polar. The best rotation pose candidate was chosen by one of the highest similarity measures returned by the methods in these domains. It has been noted in literature that frequency domain methods are robust against noise and occlusion which was also validated by the authors' results. 3D translation of the volume was then recovered by distance-map based BFGS optimization well suited to convex structure of the authors' objective function without local minima due to distance maps. A novel automatic x-ray vessel segmentation was also performed in this study. Final results were evaluated in 2D projection space for patient data; and with ground truth values and landmark distances for the images acquired with a solid phantom vessel. Results validate that rotation recovery in frequency domain is robust against differences in segmentations in two modalities. Distance-map translation is successful in aligning coronary trees with highest possible overlap. Numerical and qualitative results show that single view rigid alignment in projection space is successful. This work can be extended with multiple views to resolve depth ambiguity and with deformable registration to account for nonrigid motion in patient data.


Removal of subsurface fluorescence in cryo-imaging using deconvolution.
We compared image restoration methods [Richardson-Lucy (RL), Wiener, and Next-image] with measured "scatter" point-spread-functions, for removing subsurface fluorescence from section-and-image cryo-image volumes. All methods removed haze, delineated single cells from clusters, and improved visualization, but RL best represented structures. Contrast-to-noise and contrast-to-background improvement from RL and Wiener were comparable and 35% better than Next-image. Concerning detection of labeled cells, ROC analyses showed RL ≈Wiener > Next-image >> no processing. Next-image was faster than other methods and less prone to image processing artifacts. RL is recommended for the best restoration of the shape and size of fluorescent structures.


Anti-D prophylaxis: past, present and future.
The new British Committee for Standards in Haematology (BCSH) guidelines for the use of anti-D immunoglobulin in pregnancy provide a welcome clarification of the use of anti-D in ectopic pregnancy and after red cell salvage during caesarean section, of dosing with different preparations and distinguishing non-immune and immune anti-D. The routine use of anti-D prophylaxis (RAADP) to prevent Rhesus (Rh) D alloimmunisation during the third trimester is well established and requires careful and well-audited local implementation to achieve the maximum public health benefit. In the UK, such scrutiny may be provided by the reporting of failed anti-D prophylaxis at women who have produced an immune anti-D that is detectable for the first time in the current pregnancy through the voluntary Serious Hazards of Transfusion reporting scheme (SHOT). Application of fetal RHD genotyping would avoid giving anti-D to RhD negative women carrying an RhD negative fetus. RAADP is directed by fetal RHD genotyping in some countries in Northern Europe led by the Netherlands and Denmark. The economic case for RAADP directed by fetal RHD genotyping needs to be carefully evaluated and in England is under consideration by National Institute for Health and Clinical Excellence (NICE). Possible future developments include the use of monoclonal anti-D preparations, now in advanced clinical trials, and also testing the hypothesis that directed RAADP from early in the second trimester may further reduce anti-D immunisation.


Anti-alpha-toxin monoclonal antibody and antibiotic combination therapy improves disease outcome and accelerates healing in a Staphylococcus aureus dermonecrosis model.
Alpha-toxin (AT) is a major virulence determinant in Staphylococcus aureus skin and soft tissue infection models. We previously demonstrated that prophylactic administration of 2A3, an AT-neutralizing monoclonal antibody (MAb), prevents S. aureus disease in a mouse dermonecrosis model by neutralizing AT-mediated tissue necrosis and immune evasion. In the present study, MEDI4893*, an affinity-optimized version of 2A3, was characterized for therapeutic activity in the dermonecrosis model as a single agent and in combination with two frontline antibiotics, vancomycin and linezolid. MEDI4893* postinfection therapy was found to exhibit a therapeutic treatment window similar to that for linezolid but longer than that for vancomycin. Additionally, when combined with either vancomycin or linezolid, MEDI4893* resulted in reduced tissue damage, increased neutrophil and macrophage infiltration and abscess formation, and accelerated healing relative to those with the antibiotic monotherapies. These data suggest that AT neutralization with a potent MAb holds promise for both prophylaxis and adjunctive therapy with antibiotics and may be a valuable addition to currently available options for the treatment of S. aureus skin and soft tissue infections.


A Comparison Study of Cognitive and Neuropsychiatric Features of Essential Tremor and Parkinson's Disease.
Essential tremor (ET) and Parkinson's disease (PD) are two of the most common movement disorders. Leaving aside their motor features, these two conditions share several non-motor features, including cognitive dysfunction and personality changes. However, there are few data comparing the cognitive and personality profiles of ET with PD. Here we compare the cognitive and personality profiles of the two diseases. Thirty-two consecutive non-demented ET patients (13 females and 19 males) (67.7±9.8 years), 32 non-demented PD patients (13 females and 19 males) (67.7±9.5 years), and 32 healthy matched controls (14 females and 18 males) (67.9±10.1 years) underwent a neuropsychological test battery, including a global cognitive assessment and tests of attention, executive function, memory, language, and visuospatial function, as well as the Personality Assessment Inventory. Multivariable linear regression analyses were performed, adjusted for age, sex, years of education, medications that potentially affect cognitive function, number of medications, and the 17-item Hamilton Depression Rating Scale Total Score. Neuropsychological scores were similar in PD and ET patients, but patients with disease performed more poorly than control subjects in cognitive tasks such as attention, executive function, memory, and naming. ET and PD exhibited similar deficits in specific aspects of neuropsychological functioning, particularly those thought to rely on the integrity of the prefrontal cortex, and this suggests involvement of frontocerebellar circuits. These findings further challenge the traditional view of ET as a benign and monosymptomatic disorder.


Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation.
High-throughput mRNA sequencing (RNA-Seq) promises simultaneous transcript discovery and abundance estimation. However, this would require algorithms that are not restricted by prior gene annotations and that account for alternative transcription and splicing. Here we introduce such algorithms in an open-source software program called Cufflinks. To test Cufflinks, we sequenced and analyzed >430 million paired 75-bp RNA-Seq reads from a mouse myoblast cell line over a differentiation time series. We detected 13,692 known transcripts and 3,724 previously unannotated ones, 62% of which are supported by independent expression data or by homologous genes in other species. Over the time series, 330 genes showed complete switches in the dominant transcription start site (TSS) or splice isoform, and we observed more subtle shifts in 1,304 other genes. These results suggest that Cufflinks can illuminate the substantial regulatory flexibility and complexity in even this well-studied model of muscle development and that it can improve transcriptome-based genome annotation.


Expression analysis of p16 and TOP2A protein biomarkers in cervical cancer lesions and their correlation with clinico-histopathological characteristics in a referral hospital, Tanzania.
Biomarkers yield important information for early diagnosis of cervical cancer. However, they are rarely applied for prognosis of cervical cancer in Tanzania, where visual inspection assay with acetic acid or Lugol's iodine and Pap test are being used as the standard screening/ diagnostic methods. This was a retrospective hospital-based cross-sectional study that was conducted to assess cyclin-dependent kinase inhibitor (p16) and topoisomerase II-alpha (TOP2A) proteins expression among women seeking cervical cancer care at Kilimanjaro Christian Medical Centre, Tanzania between May 1, 2017 and May 10, 2018. Immunohistochemistry technique was used to detect the expressions of p16 and TOP2A proteins from the retrieved formalin-fixed and paraffin-embedded (FFPE) cervical biopsies. A total of 145 patients, with a mean age of 52.1 ± 12.9 years, were included in this study. Upon immunohistochemistry staining, 103 (71.0%) and 90 (62.1%) were p16 and TOP2A positive respectively. There was a strong association between histopathological class and p16/TOP2A expression levels (Fisher's exact test, p<0.001). Moreover, there was a strong positive correlation between p16/TOP2A and cancerous cervical lesions (Spearman's rank correlation coefficients = 0.833 and 0.687, p = 0.006 and 0.005, respectively). The age-adjusted odds ratio for predicting cervical cancer lesions were independently significant for p16/TOP2A biomarkers in FFPE cervical tissues [p16: OR = 1.142 (95% CI: 1.059-1.232, p<0.001) and TOP2A: OR = 1.046 (95% CI: 1.008-1.085, p = 0.015)]. Importantly, the diagnostic performance of p16 was higher than that of TOP2A in the diagnosis of cancerous lesions from non-cancerous cervical lesions (sensitivity: 97.2% versus 77.6%, accuracy: 92.8% versus 87.8%, respectively). Our study has highlighted that over-expression of TOP2A is related to the grade of cervical intraepithelial neoplasia but does not predict prognosis in cervical cancer. Similarly, expression of p16 is related to degree of histological dysplasia and malignancy, suggesting its prognostic and predictive value in the management of cervical cancers. Further bigger studies are needed to validate their applications in the early diagnosis of cervical cancer.


The expressive elaboration of imaginative narratives by children with specific language impairment.
This study investigated the expressive elaboration of narratives from children with specific language impairment (SLI). Forty-eight 6- and 8-year-old children with SLI were compared with forty-eight 6- and 8-year-old typical language (TL) children. Two imaginative narratives were scored for 14 elements of expressive elaboration in 3 categories. A subset of simple elements was analyzed separately. The effect of adult models and context was also considered. Children with SLI (whether 6 or 8 years of age) and younger TL children produced stories with significantly fewer appendages (e.g., Abstract, Coda), orientations (e.g., name, personality feature), and evaluations (e.g., interesting modifier, dialogue) than older TL children. The children with SLI and younger children showed significantly poorer performance even on simple elements such as character names and repetition (He ran and ran). Children with SLI, although performing lower than their TL age peers, demonstrated improvements from the 1st to the 2nd fictional story. Children with SLI were not differentially affected by the adult models. This study shows that expressive elaboration of narratives is sensitive to age- and language-level differences. The results suggest that children with SLI need guidance on artful storytelling, even for simple story elements.


SU-E-T-385: Accelerated Beam Delivery with MLC Gaps in IMRT Fields.
By default, the 4-mm wide leaf-pairs of the Elekta Beam Modulator MLC not used for field shaping are moved to under the primary collimator to minimize the leakage dose. The trade-off is the increased beam delivery time in the case of IMRT. This study examines the dosimetric impact and improved delivery efficiency by allowing the unused leaf-pair as 2mm gap to stay within the PTV aperture during beam-on. The capability of the treatment planning system (Pinnacle) in modeling the central and off-axis dose through 2mm MLC gaps, respectively, was investigated. EBT film measurement performed in a solid water phantom was compared with the computed dose. For two representative IMRT cases, the dosimetric features of treatment plans generated with and without MLC gaps in the field were compared. Similarly, the delivery times for 13 IMRT cases with and without MLC gaps were compared. The peak dose of the leaf gap computed with Pinnacle was dependent on dose grid resolution. The difference between a 0.5mm and 2mm dose grid was about 16.5% for 6MV and 15MV beam respectively. Using a 0.5mm dose grid, the maximum difference between measured and calculated dose was < 1% for a central axis gap, and < 3% for an off-axis gap. The maximum dose through a 2mm×4mm leaf gap was estimated < 10cGy for a typical IMRT plan. DVH comparison showed an increase of target dose up to 2% for the IMRT plans with gap, with minimal dose change to all critical structures. For the 13 IMRT cases, the beam delivery times were reduced by 27+/-9.7%, from 8.7+/-4.5 minutes to 6.5+/-4.0 minutes. The beam delivery is accelerated by utilizing minimal gaps in the IMRT fields. The increased dose to the target is acceptable and is minimal to all critical structures.


The representation of stimulus familiarity in anterior inferior temporal cortex.
1. The inferior temporal (IT) cortex plays an important role in both short- and long-term memory for visual patterns. Most previous studies of IT neurons have tested their responses in recency memory tasks, which require that the memory lasts only the length of a single behavioral trial, which may be < 1 s. To determine the role of IT neurons in longer lasting memories, we measured their responses to initially novel stimuli as the stimuli gradually became familiar to the animal. 2. Two rhesus monkeys were trained on a delayed matching to sample (DMS) task with several intervening stimuli between the sample and the final matching stimulus on each trial. The purpose of the task was to ensure that the animal attended to the stimuli and held them in memory, at least temporarily. Unlike in several previous studies, the focus was not on within-trial effects but rather on the incidental memories that built up across trials as the stimuli became familiar. Each cell was tested with a set of 20 novel stimuli (digitized pictures of objects) that the monkey had not seen before. These stimuli were used in a fixed order over the course of an hour-long recording session, and the number of intervening trials between repetitions of a given sample stimulus was varied. 3. The responses of about one-third of the cells recorded in anterior-ventral IT cortex declined systematically as the novel stimuli became familiar. After six to eight repetitions, responses reached a plateau that was approximately 40% of the peak response. Virtually all of these cells also showed selectivity for particular visual stimuli and thus were not "novelty detectors" in the sense of cells that respond to any novel stimulus. Rather, the responses of these cells were a joint function of familiarity and specific object features such as shape and color. A few cells showed increasing responses with repetition over the recording session, but these changes were accompanied by changes in baseline firing rate, suggesting that they were caused by nonspecific effects. 4. The decrement in response with familiarity was stimulus specific and bridged > 150 presentations of other stimuli, the maximum tested. For some cells the maximum decrement in response occurred for those stimuli that initially elicited the largest response. There was no significant change in response to stimuli that were already familiar. 5. The same cells that showed familiarity effects also showed reduced responses to the matching stimuli at the end of each trial, compared with the responses to the samples.(ABSTRACT TRUNCATED AT 400 WORDS)


The effect of task and pitch structure on pitch-time interactions in music.
Musical pitch-time relations were explored by investigating the effect of temporal variation on pitch perception. In Experiment 1, trained musicians heard a standard tone followed by a tonal context and then a comparison tone. They then performed one of two tasks. In the cognitive task, they indicated whether the comparison tone was in the key of the context. In the perceptual task, they judged whether the comparison tone was higher or lower than the standard tone. For both tasks, the comparison tone occurred early, on time, or late with respect to temporal expectancies established by the context. Temporal variation did not affect accuracy in either task. Experiment 2 used the perceptual task and varied the pitch structure by employing either a tonal or an atonal context. Temporal variation did not affect accuracy for tonal contexts, but did for atonal contexts. Experiment 3 replicated these results and controlled potential confounds. We argue that tonal contexts bias attention toward pitch and eliminate effects of temporal variation, whereas atonal contexts do not, thus fostering pitch-time interactions.


The identification and psychological treatment of panic disorder in adolescents: a survey of CAMHS clinicians.
Panic disorder is experienced by around 1% of adolescents and has a significant impact on social and academic functioning. Preliminary evidence supports the effectiveness of panic disorder-specific treatment in adolescents with panic disorder; however, panic disorder may be overlooked in adolescents due to overlapping symptoms with other anxiety disorders and other difficulties being more noticeable to others. The aim of this study was to establish what training National Health Service (NHS) Child and Adolescent Mental Health Services (CAMHS) clinicians have received in psychological therapies and panic disorder and how they identify and treat panic disorder in adolescents. CAMHS clinicians from a range of professions (n = 427), who were delivering psychological treatments to children and adolescents with anxiety disorders, participated. They completed a cross-sectional, online survey, including a vignette describing an adolescent with panic disorder, and were asked to identify the main diagnosis or presenting problem. Less than half the clinicians (48.6%) identified panic disorder or panic symptoms as the main presenting problem from the vignette. The majority of clinicians suggested CBT would be their treatment approach. However, few identified an evidence-based treatment protocol for working with young people with panic disorder. Almost half the sample had received no training in cognitive behaviour therapy (CBT), and around a fifth had received no training in delivering psychological treatments. Only half of CAMHS clinicians identified panic disorder from a vignette and although CBT treatments are widely offered, only a minority of adolescents with panic disorder are receiving treatments developed for and evaluated with young people with panic disorder. There is a vital need for clinician training, the use of tools that aid identification and the implementation of evidence-based treatments within CAMHS. Panic disorder affects around 1% of adolescents, adversely impacting social, academic and long-term life functioning. There is preliminary evidence for the effectiveness of a panic disorder-specific treatment of panic disorder in adolescents. Clinicians struggle to identify panic disorder or suitable treatment protocols for treating adolescents, although most would use CBT as the treatment approach. There is a vital need for clinician training, tools that aid identification of young people with panic disorder and evidence-based treatments that can be delivered in routine clinical settings.


Sudden infant death syndrome and cardiac arrhythmias.
There is a considerable body of evidence that common bacterial toxins, absorbed from the mucosal surface or delivered as part of a transient bacteremia, have a pathogenic role in sudden infant death syndrome (SIDS). The candidate organisms are Staphylococcus aureus and Escherichia coli. Death in SIDS is rapid, with infants progressing from well, or only mildly unwell, to death in less than 20 min. This mode of death is not typical of infection but it is consistent with toxin action on cardiovascular or respiratory control. Both S. aureus and E. coli secrete toxins (cytolysins and colicins) that create channels in cell membranes and disturb ion currents. Recent evidence indicates that between 5 and 15% of SIDS cases carry potentially lethal loss-of-function mutations in cardiac channelopathy genes. However, only a minority of individuals with these mutations die of SIDS and the hypothesis proposed is that toxin-gene interaction could explain the deaths. Furthermore, channelopathy mutations predispose to sudden death at all ages and since episodes of transient bacteremia occur throughout life the idea of toxin-gene interaction could have wider applicability. These ideas can be investigated and answered in the near future using the new science of proteomics.


Descriptive analysis of common functional limitations identified by patients with shoulder pain.
Recent establishment of G-codes by the US government requires therapists to report function limitations at initial evaluation. Limited information exists specific to the most common limitations in patients with shoulder pain. To describe the most commonly expressed shoulder limitations with activities and their severity/level of impairment from a patient's perspective on the initial evaluation. Descriptive. Patients reporting pain with overhead activity and seeking medical attention from one orthopedic surgeon were recruited as part of a cohort study. 176 with shoulder superior labral tear from anterior to posterior (SLAP), subacromial impingement, combined SLAP and rotator cuff, and nonspecific (female = 53, age = 41 ± 13 y; male = 123, age = 41 ± 12 y). Data were obtained on the initial visit from the Patient-Specific Functional Scale (PSFS) questionnaire. Three researchers extracted meaningful concepts from the PSFS and linked them to the International Classification of Functioning (ICF) categories according to established ICF linking rules. 176 participants yielded 765 meaningful concepts that were linked to the ICF with a 66% agreement between researchers before consensus. There were no differences between diagnoses. Of all patients, 88% reported functional limitations coded into meaningful concepts as represented by 10 ICF codes; 634 (83%) meaningful concepts were linked to the activities and participation domain while 129 (17%) were linked to the body function domain. Only 2 reported functional limitations that were considered nondefinable (nd). The overall average initial impairment score on the PSFS = 4 ± 2.5 out of 10 points. Meaningful concepts from the activities and participation domain were most commonly identified as functional limitations and were more prevalent than limitations from the body function domain. This information helps identify some of the most common limitations in patients with shoulder pain that therapists can use to efficiently document patient functional impairment.


Review: intellectual property aspects of plant transformation.
One of the recurring themes of the debates concerning the application of genetic transformation technology has been the role of Intellectual Property Rights (IPR). This term covers both the content of patents and the confidential expertise usually related to methodology and referred to as 'Trade Secrets'. This review explains the concepts behind patent protection, and discusses the wide-ranging scope of existing patents that cover all aspects of transgenic technology, from selectable markers and novel promoters to methods of gene introduction. Although few of the patents in this area have any real commercial value, there are a small number of key patents that restrict the 'freedom to operate' of new companies seeking to exploit the methods. Over the last 20 years, these restrictions have forced extensive cross-licensing between ag-biotech companies and have been one of the driving forces behind the consolidation of these companies. Although such issues are often considered of little interest to the academic scientist working in the public sector, they are of great importance in any discussion of the role of 'public-good breeding' and of the relationship between the public and private sectors.