The perils of global signal regression for group comparisons: a case study of Autism Spectrum Disorders.
We have previously argued from a theoretical basis that the standard practice of regression of the Global Signal from the fMRI time series in functional connectivity studies is ill advised, particularly when comparing groups of participants. Here, we demonstrate in resting-state data from participants with an Autism Spectrum Disorder and matched controls that these concerns are also well founded in real data. Using the prior theoretical work to formulate predictions, we show: (1) rather than simply altering the mean or range of correlation values amongst pairs of brain regions, Global Signal Regression systematically alters the rank ordering of values in addition to introducing negative values, (2) it leads to a reversal in the direction of group correlation differences relative to other preprocessing approaches, with a higher incidence of both long-range and local correlation differences that favor the Autism Spectrum Disorder group, (3) the strongest group differences under other preprocessing approaches are the ones most altered by Global Signal Regression, and (4) locations showing group differences no longer agree with those showing correlations with behavioral symptoms within the Autism Spectrum Disorder group. The correlation matrices of both participant groups under Global Signal Regression were well predicted by our previous mathematical analyses, demonstrating that there is nothing mysterious about these results. Finally, when independent physiological nuisance measures are lacking, we provide a simple alternative approach for assessing and lessening the influence of global correlations on group comparisons that replicates our previous findings. While this alternative performs less well for symptom correlations than our favored preprocessing approach that includes removal of independent physiological measures, it is preferable to the use of Global Signal Regression, which prevents unequivocal conclusions about the direction or location of group differences.


Gender differences in morningness-eveningness preference.
Morningness-eveningness preference (morning-, intermediate-, evening-type) or circadian typology is the individual difference that most clearly explains the variations in the rhythmic expression of biological or behavioral patterns. The aim of this study was to analyze gender difference in morningness-eveningness preference using the Horne and Ostberg questionnaire in the largest university student population selected so far (N = 2135), with an age range 18-30 yr. Morningness-eveningness questionnaire (MEQ) score distribution closely correlated to the normal curve (range 17-78, mean = 48.25; SD = 10.11), with 338 (15.84%) morning-types, 1273 (59.62%) intermediate-types, and 524 (24.54%) evening-types. The men and women differed significantly in their mean scores (p < 0.0001) and distribution per circadian typology (p < 0.00001), with the men presenting a more pronounced eveningness preference. Three factors were identified by factor analysis: time of greatest efficiency (I), sleep time/sleep phase (II), awakening time/sleep inertia (III). The MEQ items sensitive to gender differences were essentially those included in factor I and factor II. The results are discussed in relation to recent models of circadian regulation of the sleep-wake cycle.


Simulations of host-plasmid interactions in Escherichia coli: Copy number, promoter strength, and ribosome binding site strength effects on metabolic activity and plasmid gene expression.
A mechanistically detailed single-cell model E. coli B/r-A was adapted to simulate the effects of vector presence on cell metabolism. Competition for RNA polymerase between chromosome and plasmid DNA is explicitly included. Distribution of active ribosomes among chromosome- and plasmid-derived messenger RNA, another key facet of host-plasmid interactions, is also treated in detail. Simulations of recombinant cell growth rate and cloned-gene productivity as a function of relative plasmid number per cell agree closely with experimental results. Model prediction of the variation of cell cycle parameters C and D with plasmid number are roughly consistent with available data. Models of this class can be used to simulate changes in productivity resulting from specific alterations in the expression vector. The effects of changing cloned-gene promoter and ribosome binding strengths and of augmenting cell transcription or translation capacity have been studied using the recombinant cell model. Results suggest that cloned-gene expression is limited by cellular transcription capacity. These and other parametric studies, conveniently implemented using the computer cell, provide important guidance for future experiments directed at better understanding of host-plasmid interactions and at optimizing recombinant system productivity.


Effect of a low-fat diet enriched either with rapeseed oil or sunflower oil on plasma lipoproteins in children and adolescents with familial hypercholesterolaemia. Results of a pilot study.
There is convincing evidence that unsaturated fatty acids exert favourable effects on plasma cholesterol levels. However, it is not clear which type of oil has the most pronounced effect, especially not in paediatric patients. The aim was to compare two low-fat diet regimes enriched with either monounsaturated fatty acids by rapeseed oil (RO) or polyunsaturated fatty acids by sunflower oil (SO) in children affected with familial hypercholesterolaemia (FH). Twenty-one children aged 6-18 years affected with FH were enrolled in this randomized and double-blind pilot trial. The subjects and their families were trained to adhere to a low-fat/low-cholesterol diet. All visible fats were to be replaced by either RO or SO (14-27 g/day) for 13 weeks. Dietary adherence was controlled by repeated 4-day dietary records; plasma lipids, lipoproteins and risk markers were assessed at baseline and post-intervention. Out of 21 subjects, 16 could be followed-up after 6 months. Both fat-modified diets resulted in significant reduction in total cholesterol concentrations of 9.4% (RO P<0.005 vs SO P<0.05) and low-density lipoprotein (LDL) cholesterol concentrations of 12.7% (P<0.005) for RO and 11.3% (P<0.05) for SO. The reduction of the LDL/high-density lipoprotein (HDL) cholesterol ratio (RO 9% vs SO 3.5%) and high-sensitivity C-reactive protein (RO 16.8% vs SO 1.7%) were not statistically significant, respectively. In most participating families, a change in eating habits could be observed. A fat-modified diet enriched with RO seems to have very similar effects on cholesterol levels as with SO. However, our study suggests that RO has possibly more favourable effects concerning cardiovascular risk profile. Both diets appear to be feasible and were well accepted among our subjects. Although these results are promising, larger trials will be required to validate our findings.


Involvement of phosphatidylinositol 3-kinase in NFAT activation in T cells.
Phosphatidylinositol 3-kinase (PI3-K) has been implicated in the regulation of cell proliferation in many cell types. We have previously shown that in T cells the PI3-K inhibitor, wortmannin, interferes with activation of the mitogen-activated kinase, Erk2, after T cell receptor (TcR) stimulation. To further explore the involvement of PI3-K in T cell activation, we created a set of potentially dominant negative PI3-K constructs comprising individual or tandem domains of the regulatory p85 subunit and tested their effect on downstream signaling events like Erk2 activation and transcription from an NFAT (nuclear factor of activated T cells) element taken from the interleukin-2 promoter. Following TcR stimulation, activation of Erk2 was only inhibited by a previously described truncated form of p85 that cannot bind the catalytic subunit, but not by other constructs of p85. In contrast, several mutant p85 alleles had dramatic effects on NFAT activation. Most interestingly, the N-terminal SH2 domain had an inhibitory effect, whereas a mutant p85 containing only the two SH2 domains enhanced basal NFAT activity in a Ras-dependent manner. Ionomycin induced synergistic activation of NFAT in cells expressing p85 mutants that contained the C-terminal SH2 domain. Analysis of phosphotyrosine-containing proteins bound to truncated p85 constructs revealed cooperative binding of the two SH2 domains but no apparent differences between the N- and C-terminal SH2 domains. Wortmannin did not interfere with NFAT activation, although it inhibited PI3-K and Erk2 activation in the same experiment. These results suggest that PI3-K is involved in NFAT activation through a complex adaptor function of its regulatory subunit and that its lipid kinase activity is dispensable for this effect.


Secretory dysfunction of vascular endothelium limits the effect of angiotensin converting enzyme inhibitor quinapril on aggregation of erythrocytes in experimental hypertension.
Using automatic erythrocyte aggregometer type MA-1 (Myrenne gmbh, Germany), we investigated the hypothesis that therapeutic effectiveness of quinapril--angiotensin converting enzyme inhibitor (ACEI)--in the treatment of hypertension would correlate with improvement of red blood cell (RBC) aggregability. Experiments were performed on commercially available inbred strain of spontaneously hypertensive male rats (SHR) aged 19-21 weeks. Age-matched normotensive Wistar-Kyoto (WKY) rats genetically related to SHR were used as a control. Aggregability of RBC in hypertensive rats was significantly higher than in control WKY animals. Quinapril (100 microg/kg) administered i.p. for 8 days improved RBC aggregability in normotensive rats but surprisingly not in SHR animals. Beneficial effect of quinapril on RBC aggregation observed in normotensive animals did not occur when this drug was injected in combination with aspirin (1 or 50 mg/kg) or with indomethacin (20 mg/kg) or with L-NAME (10 mg/kg). However, much the same damaging effects on RBC aggregability were observed when aspirin, indomethacin or L-NAME were each administered into normotensive animals without quinapril. In contrast with normotensive rats, aggregability of RBC in SHR was not affected either by quinapril or by indomethacin and by L-NAME, given separately or in combination. The only compound significantly worsening RBC aggregability in SHR was aspirin but this effect was not dose-dependent. Quinapril-induced improvement of RBC aggregability in normotensive rats (but not in SHR) was completely abolished by simultaneous administration of B2 receptor antagonist icatibant and successfully mimicked by 8 days of treatment with bradykinin. In vitro aggregability of RBC isolated from WKY was not affected by previous incubation (30 min at 37 degrees C) with quinapril, indomethacin or L-NAME. Only aspirin (3 mM) significantly increased RBC aggregability as compared to placebo. It is concluded that under physiological conditions quinapril efficiently inhibits RBC aggregability and this effect is modulated by secretion of endothelial mediators, mainly prostacyclin and nitric oxide. In hypertension quinapril, in spite of lowering of arterial blood pressure, is unable to display its beneficial effects on RBC aggregability possibly due to the hypertension-induced/accompanied dysfunction of vascular endothelium. Aspirin revealed unique erythrocyte damaging properties, presumably independent of inhibition of cyclooxygenase but related to a direct membrane protein acetylation.


Eicosanoids and the esophagus.
Eicosanoids are metabolites of arachidonic acid and include, among other compounds, the prostaglandins and leukotrienes. These metabolites are known to play important roles in the health and disease of many gastrointestinal tissues. Less attention, however, has been given to their functions in the esophagus. The present study, therefore, aims both to review the current knowledge about eicosanoids in healthy and diseased esophagi, and to critically assess the data pertaining to their role in the pathophysiology of gastroesophageal reflux disease.


Influence of ligand valency on the targeting of immature human dendritic cells by mannosylated liposomes.
An important challenge for the development of new generations of vaccines is the efficient delivery of antigens to antigen presenting cells such as dendritic cells. In the present study we compare the interaction of plain and targeted liposomes, containing mono-, di-, and tetraantennary mannosyl lipid derivatives, with human monocyte-derived immature dendritic cells (iDCs). Whereas efficient mannose receptor-mediated endocytosis by iDCs was observed for the mannosylated liposomes, in contrast, only nonspecific interaction with little uptake was observed with plain liposomes. In accordance with the clustering effect, liposomes prepared with multibranched mannosylated lipids displayed higher binding affinity for the mannose receptor than vesicles containing the monomannosylated analogs. Importantly, we have found that dimannosylated ligands present at the surface of the liposomes were as efficient as tetramannosylated ones to engage in multidentate interactions with the mannose receptor of iDCs, resulting in both cases in an effective uptake/endocytosis. This result will greatly facilitate, from a practical standpoint, the design of mannose-targeted vaccination constructs. Moreover, we showed that mannose-mediated uptake of liposomes did not result in an activation of iDCs. Altogether, our results suggest that antigen-associated targeted liposomes containing diantennary mannosylated lipids could be effective vectors for vaccines when combined with additional DC activation signals.


Depression and anxiety disorders in chronic hemodialysis patients and their quality of life: A cross-sectional study about 106 cases in the northeast of morocco.
Hemodialysis (HD) has a severe impact on the life of HD patients. The aim of this work was to assess the prevalence of depression and anxiety disorders, suicidal ideation, and the quality of life among HD patients. Associated factors were also studied. A cross-sectional study was carried out among 103 HD patients treated at the HD Center of Al Farabi Hospital of Oujda during a period of six months in 2015. The Mini-International Neuropsychiatric Interview and European Quality of Life-5 Dimensions (EQ-5D) were used for the assessment. Major depressive episode (MDE) was found in 34% of our patients, whereas anxiety disorder was observed in 25.2%. Suicidal ideation was found in 16.5% and 1.9% of our patients planned their suicide. The EQ-5D index was 0.41 ± 0.36 and the EQ-Visual Analog Scale score was 45.73 ± 14. Multivariate analysis showed that MDEs were associated with three factors: marital status, pain, and anxiety disorder. There was also an association between anxiety disorder and age and EQ-Visual Analog Scale score. Suicidal ideation was associated with marital status and anxiety disorders. Together, these results underline the importance of the collaboration between nephrologists and psychiatrists for a better care of HD patients.


Three-year outcome with drug-coated balloon percutaneous coronary intervention in coronary Takayasu arteritis: A case review.
We describe the first long-term follow-up of a young patient with active Takayasu arteritis who presented with an acute coronary syndrome, treated endovascularly with percutaneous coronary intervention without stenting. A drug-coated balloon was used with high-resolution coronary imaging guidance in the form of optical coherence tomography on a critical ostial left anterior descending coronary artery lesion. A repeat procedure was undertaken after 4 months confirming a durable coronary angioplasty result and the patient remained symptom-free beyond 3 years. Coronary stenting in this population is associated with early and aggressive stent failure. Hence, this is an innovative approach. We believe that the stent, regardless of whether it is first, second or subsequent generation, leaves a permanent foreign body within the vasculature that becomes the seed for inflammatory reactions, resulting in recurrent in-stent restenotic fibrosis irrespective of concurrent immunotherapy or the degree of disease activity.


Health hazards of farming.
The U.S. farm work force numbers approximately 6.5 million. The health risks connected with their farm tasks are many and varied. Accidents and illnesses are caused by tractors, specialized farming equipment, chemicals, zoonoses, fungi, sensitizers, and the environment of confinement buildings. The scattered nature of the work force makes preventive measures difficult to implement.


Austrian Syndrome - A Devastating Osler's Triad: Case Report and Literature Review.
Austrian syndrome is a rare triad of meningitis, pneumonia, and endocarditis caused by Streptococcus pneumoniae. We present a case of the Austrian syndrome in the oldest patient in the reviewed literature, with no other classically described risk factors. She had an unusual initial presentation and microorganism portal of entry. Her hospital course was complicated by the diagnosis of monoclonal gammopathy, septic knee joint, septic brain emboli and respiratory failure. We also provide an extensive review of available literature of this commonly unrecognized entity.


Beneficial Effects of Probiotic Consumption on the Immune System.
The gastrointestinal tract is one of the most microbiologically active ecosystems that plays a crucial role in the working of the mucosal immune system (MIS). In this ecosystem, the consumed probiotics stimulate the immune system and induce a network of signals mediated by the whole bacteria or their cell wall structure. This review is aimed at describing the immunological mechanisms of probiotics and their beneficial effects on the host. Once administered, oral probiotic bacteria interact with the intestinal epithelial cells (IECs) or immune cells associated with the lamina propria, through Toll-like receptors, and induce the production of different cytokines or chemokines. Macrophage chemoattractant protein 1, produced by the IECs, sends signals to other immune cells leading to the activation of the MIS, characterized by an increase in immunoglobulin A+ cells of the intestine, bronchus and mammary glands, and the activation of T cells. Specifically, probiotics activate regulatory T cells that release IL-10. Interestingly, probiotics reinforce the intestinal barrier by an increase of the mucins, the tight junction proteins and the Goblet and Paneth cells. Another proposed mechanism of probiotics is the modulation of intestinal microbiota by maintaining the balance and suppressing the growth of potential pathogenic bacteria in the gut. Furthermore, it has been demonstrated that long-term probiotics consumption does not affect the intestinal homeostasis. The viability of probiotics is crucial in the interaction with IECs and macrophages favoring, mainly, the innate immune response. Macrophages and Dendritic cells (DCs) play an important role in this immune response without inducing an inflammatory pattern, just a slight increase in the cellularity of the lamina propria. Besides, as part of the machinery that probiotics activate to protect against different pathogens, an increase in the microbicidal activity of peritoneal and spleen macrophages has been reported. In malnutrition models, such as undernourishment and obesity, probiotic was able to increase the intestinal and systemic immune response. Furthermore, probiotics contribute to recover the histology of both the intestine and the thymus damaged in these conditions. Probiotic bacteria are emerging as a safe and natural strategy for allergy prevention and treatment. Different mechanisms such as the generation of cytokines from activated pro-T-helper type 1, which favor the production of IgG instead of IgE, have been proposed. Key Messages: Probiotic bacteria, their cell walls or probiotic fermented milk have significant effects on the functionality of the mucosal and systemic immune systems through the activation of multiple immune mechanisms.


Early diagnosis for the onset of peri-implantitis based on artificial neural network.
The aim of this study is to construct an artificial neural network (ANN) based on bioinformatic analysis to enable early diagnosis of peri-implantitis (PI). PI-related datasets were retrieved from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) and functional enrichment analyses were performed between PI and the control group. Furthermore, the infiltration of 22 immune cells in PI was analyzed using CIBERSORT. Hub genes were identified with random forest (RF) classification. The ANN model was then constructed for early diagnosis of PI. A total of 1,380 DEGs were identified. Enrichment analysis revealed the involvement of neutrophil-mediated immunity and the NF-kappa B signaling pathway in PI. Additionally, higher proportion of naive B cells, activated memory CD4 T cells, activated NK cells, M0 macrophages, M1 macrophages, and neutrophils were observed in the soft tissues surrounding PI. From the RF analysis, 13 hub genes (ST6GALNAC4, MTMR11, SKAP2, AKR1B1, PTGS2, CHP2, CPEB2, SYT17, GRIP1, IL10, RAB8B, ABHD5, and IGSF6) were selected. Subsequently, the ANN model for early diagnosis of PI was constructed with high performance. We identified 13 hub genes and developed an ANN model that accurately enables early diagnosis of PI.


Measurement of SARS-CoV-2 Antibody Titers Improves the Prediction Accuracy of COVID-19 Maximum Severity by Machine Learning in Non-Vaccinated Patients.
Numerous studies have suggested that the titers of antibodies against SARS-CoV-2 are associated with the COVID-19 severity, however, the types of antibodies associated with the disease maximum severity and the timing at which the associations are best observed, especially within one week after symptom onset, remain controversial. We attempted to elucidate the antibody responses against SARS-CoV-2 that are associated with the maximum severity of COVID-19 in the early phase of the disease, and to investigate whether antibody testing might contribute to prediction of the disease maximum severity in COVID-19 patients. We classified the patients into four groups according to the disease maximum severity (severity group 1 (did not require oxygen supplementation), severity group 2a (required oxygen supplementation at low flow rates), severity group 2b (required oxygen supplementation at relatively high flow rates), and severity group 3 (required mechanical ventilatory support)), and serially measured the titers of IgM, IgG, and IgA against the nucleocapsid protein, spike protein, and receptor-binding domain of SARS-CoV-2 until day 12 after symptom onset. The titers of all the measured antibody responses were higher in severity group 2b and 3, especially severity group 2b, as early as at one week after symptom onset. Addition of data obtained from antibody testing improved the ability of analysis models constructed using a machine learning technique to distinguish severity group 2b and 3 from severity group 1 and 2a. These models constructed with non-vaccinated COVID-19 patients could not be applied to the cases of breakthrough infections. These results suggest that antibody testing might help physicians identify non-vaccinated COVID-19 patients who are likely to require admission to an intensive care unit.


Study of the Vapor Phase Over Fusarium Fungi Cultured on Various Substrates.
The compositions of volatile organic compounds (VOCs) emitted by Fusarium fungi (F. langsethiae, F. sibiricum, F. poae, and F. sporotrichioides) grown on two nutritive substrates: potato sucrose agar (PSA) and autoclaved wheat kernels (WK) were investigated. The culturing of fungi and study of their VOC emissions were performed in chromatographic vials at room temperature (23 - 24 °C) and the VOCs were sampled by a solid-phase microextraction on a 85 μm carboxen/polydimethylsiloxane fiber. GC/MS was performed using a 60-m HP-5 capillary column. Components of the VOC mixture were identified by electron impact mass spectra and chromatographic retention indices (RIs). The most abundant components of the VOC mixture emitted by Fusarium fungi are EtOH, AcOH, (i) BuOH, 3-methylbutan-1-ol, 2-methylbutan-1-ol, ethyl 3-methylbutanoate, terpenes with M 136, sesquiterpenes with M 204 (a total of about 25), and trichodiene. It was found that the strains grown on PSA emit a wider spectrum and larger amount of VOCs compared with those grown on wheat kernels. F. langsethiae strain is the most active VOC producer on both substrates. The use of SPME and GC/MS also offers the potential for differentiation of fungal species and strains.


Structural basis for inhibition of the RNA-dependent RNA polymerase from SARS-CoV-2 by remdesivir.
The pandemic of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global crisis. Replication of SARS-CoV-2 requires the viral RNA-dependent RNA polymerase (RdRp) enzyme, a target of the antiviral drug remdesivir. Here we report the cryo-electron microscopy structure of the SARS-CoV-2 RdRp, both in the apo form at 2.8-angstrom resolution and in complex with a 50-base template-primer RNA and remdesivir at 2.5-angstrom resolution. The complex structure reveals that the partial double-stranded RNA template is inserted into the central channel of the RdRp, where remdesivir is covalently incorporated into the primer strand at the first replicated base pair, and terminates chain elongation. Our structures provide insights into the mechanism of viral RNA replication and a rational template for drug design to combat the viral infection.


Macromolecular crowding of molecular imprinting: A facile pathway to produce drug delivery devices for zero-order sustained release.
This paper reported the facile fabrication of drug delivery devices for zero-order sustained release by molecular crowding strategy of molecularly imprinting technology. Crowding-assisted molecularly imprinting polymers (MIPs) matrices were prepared by free-radical precipitation polymerization using aminoglutethimide (AG) as a model drug. The crowding effect was achieved by adding polystyrene as a macromolecular co-solute in pre-polymerization mixture. The MIP prepared under the non-MMC condition and the two corresponding non-imprinted particles were tested as controlled vehicles. The release profiles presented zero-order behaviors from two crowding-assisted polymers, the duration of approximately 18h for the crowding-assisted MIP and 10h for the crowding-assisted NIP, respectively while AG were all very rapid released from the other two controlled particles (85% occurring in the first hour). The BET surface area and pore volume of the crowding-assisted MIP were about ten times than those of the controlled MIP. The value of imprinting factor is 6.02 for the crowding-assisted MIP and 1.19 for the controlled MIP evaluated by the equilibrium adsorption experiment. Furthermore, the values of effective diffusivity (Deff) obtained from crowding-assisted MIP (10(-17)cm(2)/s) was about two orders of magnitude smaller than those from the controlled MIP, although the values of free drug diffusivity (D) were all found in the order of 10(-13)cm(2)/s. Compared with the commercial AG tablet, the MMC-assisted MIP gave a markedly high relative bioavailability of 266.3%, whereas the MMC-assisted NIP gave only 57.7%. The results indicated that the MMC condition can modulate the polymer networks approaciate to zero-order release of the drug and maintain the molecular memory pockets, even if under the poor polymerization conditions of MIPs preparation.


Radiographic vertebral left atrial size: A reference interval study in healthy adult dogs.
Thoracic radiography is a useful technique for the evaluation of cardiac dimensions, especially when echocardiography is not possible. The vertebral left atrial size (VLAS) has recently been proposed as a new radiographic method for quantifying left atrial dimensions in dogs. The aims of this retrospective, reference interval, observational study were to describe values for VLAS in a group of healthy adult dogs and to compare intraobserver and interobserver agreement for VLAS versus vertebral heart score (VHS) methods. Thoracic radiographs of 80 healthy adult dogs were evaluated. Normal left atrial size was determined based on an echocardiographic left atrial-to-aorta ratio < 1.6. The VLAS and vertebral heart score values were measured from right lateral radiographs. The correlations between VLAS, the vertebral heart score, and the left atrial-to-aorta ratio were evaluated. The effects of body weight, sex, and age were also tested using regression analyses. The median value of VLAS were 1.9, with a reference interval of 1.4-2.2. A positive correlation was found between VLAS and vertebral heart score values (r = 0.53; P < .0001). No effect of body weight, sex, and age on VLAS was detected. Excellent intraobserver and interobserver agreements were found for both VLAS and vertebral heart score methods (intraclass correlation coefficients ≥ 0.91). Findings from this sample of healthy dogs indicated that VLAS is a repeatable radiographic method for quantifying left atrial size. Further studies are warranted to evaluate this measure in clinically affected dogs.


Halothane and cyclopiazonic acid modulate Ca-ATPase oligomeric state and function in sarcoplasmic reticulum.
We have studied the effects of cyclopiazonic acid (CPA) and halothane on the enzymatic activity, oligomeric state, and conformational equilibrium of the Ca-ATPase in skeletal muscle sarcoplasmic reticulum (SR). CPA is a potent inhibitor of Ca-ATPase activity, and this inhibition is competitive with respect to ATP concentration. Time-resolved phosphorescence anisotropy was used to detect the fraction of Ca-ATPase monomers, dimers, and larger aggregates in the absence and presence of CPA. CPA increased the fraction of dimers and larger aggregates of the Ca-ATPase. Addition of halothane to SR, or detergent solubilization of the Ca-ATPase, increased the apparent KI of CPA inhibition, and increased the fraction of Ca-ATPase present as monomers. CPA stabilized the E2 conformational state of the Ca-ATPase relative to the E1 and E2-P states, as measured by fluorescein 5-isothiocyanate fluorescence and enzyme phosphorylation from inorganic phosphate. E2-P formation in the presence of CPA was partially restored by halothane and solubilization. We conclude that CPA inhibits the Ca-ATPase in part by overstabilizing dimers or small oligomers of the Ca-ATPase, which is correlated with stabilization of the E2 conformation of the enzyme.


Observing single nanoparticle events at the orifice of a nanopipet.
Single nanoparticle (NP) events are successfully observed at the orifice of a nanopipet by blocking the ionic current with a single NP. In addition to the traditional translocation events, we observe both staircase and blip current transients by controlling the radius ratio of NPs to nanopipet or bias potential. Confocal fluorescence microscopy and finite element simulation are used to simultaneously monitor and quantitatively understand these events, respectively. The frequency of the staircase and blip events is proportional to the NP concentration, and could be used for the quantification of NPs. This study offers a new method for NP determination and single NP behavior study.


Inversion upstream of FOXF1 in a case of lethal alveolar capillary dysplasia with misalignment of pulmonary veins.
Alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV) is a congenital malformation that leads to severe pulmonary hypertension and respiratory failure. It has been associated with deletion of, or mutation in, FOXF1 on 16q24.1, a gene encoding a forkhead transcription factor expressed in the mesenchyme of the developing lung. Here we report on the identification of a pericentric inversion on chromosome 16 (p11.2q24.1) in a case of lethal ACDMPV with atrioventricular septal defect and duodenal atresia. Array-CGH indicated that the inversion is balanced, and FISH showed that the q-arm breakpoint occurs 134 ± 10 kb upstream (5'; centromeric) of FOXF1. This is suggestive of cis-regulatory elements located more than 130 kb 5' of FOXF1, and analysis of genome-wide data sets of chromatin modifications in two different cell types suggested that the FOXF1 regulatory domain covers more than 300 kb, and perhaps up to 433 kb, upstream of the gene, but only 3 kb downstream. The 588 kb gene-free region between FOXF1 and the next gene in the centromeric direction, IRF8, is highly conserved between species and divided into two distinct regulatory domains by an insulator element. Another putative insulator occurs just downstream of FOXF1. Our results further strengthen the association between FOXF1 and a spectrum of malformations that include ACDMPV, atrioventricular septal defects, and gastrointestinal atresia. Furthermore, the presented analysis aids in defining the critical genomic region for this syndrome.


Cytokines gene expression patterns of bovine milk during middle and late stages of lactation.
The cytokine mRNA profiles of the bovine mammary gland were investigated using newly developed TaqMan real-time polymerase chain reaction systems (Applied Biosystems, Foster City, CA, USA). Transcriptional activity of six cytokines, interleukin (IL)-2, IL-6, IL-12, interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and granulocyte-macrophage colony stimulating factor (GM-CSF) was studied during the mid- and late-lactation stages. Transcripts for TNF-alpha, GM-CSF, and IFN-gamma were detected in all samples of both stages. However, IL-12 was only detected in 80 and 58 % of late- and mid-lactation samples, respectively. IL-12 expression was up-regulated in late lactation in comparison with the corresponding level in mid-lactation. The cytokines interaction in late lactation was more co-ordinated and their transcriptional levels were significantly correlated among each other, whereas, in mid-lactation significant correlation of the cytokines transcription was only seen with the TNF-alpha, GM-CSF, and IFN-gamma. Cytokine mRNA profiles between mid- and late lactation showed significant differences, which can be attributed to the dramatic changes that the mammary gland is subjected to during late lactation. The significant elevation of IL-12 transcriptional activity in late lactation and its relevance to the mammary gland immunity is discussed.


Dental reductions and dental caries.
Although first permanent molar hypoconulid absence, third molar agenesis, and small tooth size are all part of evolutionary trend of dental retardation, each bears a different relationship to dental caries. Caries prevalence in the maxillary and mandibular permanent first molars of the Burlington Research Centre serial experimental group at age 16 years was less in the children whose first molars were missing the hypoconulid. Conversely, caries prevalence in mandibular first molars was greater in the children who had agenesis of third molars. The extraction of first molars due to caries was more frequent in children with agenesis of third molars, less frequent in those with absence of hypoconulids of the first molars and unrelated to tooth size. Caries prevalence was less in small teeth, and occurred least in the small mandibular first molars with four cusps. Whereas this is in harmony with the hypothesis that evolutionary dental reductions resulted from the pressure of caries, the increased prevalence of caries and extractions coinciding with third molar agenesis does not support this view. In addition, agenesis of hypoconulids and agenesis of third molars were related to changes in structures unrelated to caries.


Perception, memory and pathological identification as precipitating factors in parental attacks on children.
This paper examines the origins and the importance of pathological identification by the parent of the child who was maltreated or is at risk of maltreatment. The understanding of these identifications suggests methods of effective preventive action without recourse to the undesirable practice of removing the child from the home, other than in exceptional circumstances. Brief case history vignettes illustrate the clinical application of this approach.


Frequency and trends of hospital discharges against medical advice (DAMA) in a large administrative database.
The aim of this research was to characterize hospitalizations associated with discharges against medical advice (DAMA) in a large, population-based data system. This was a retrospective cohort study on 11 436 500 hospital admissions. The hospital discharge records for residents of the Veneto region (north-east Italy) discharged from 2001 to 2012, from both public and accredited private hospitals, were considered. The DAMA rate was calculated by type of hospital admission, excluding patients who died. The time trend of the DAMA rate was charted from the average annual percent changes. During the period considered, 66 549 DAMA were recorded, amounting to an overall DAMA rate of 6.0‰ admissions. Analyzing the diagnostic categories, admissions for substance abuse (drugs or alcohol) and dependence coincided with the highest DAMA rate (83.5‰), followed by poisoning (40.2‰), psychiatric disorders (24.7 ‰), traumas (21.1‰), HIV-related diseases (19.9‰), burns (10.5‰), and - for women - issues relating to pregnancy, childbirth and the postnatal period (11.2‰). The DAMA rate dropped from 6.72 to 5.55 from 2000 to 2008, then remained stable. The DAMA rate dropped slightly over the period considered. Several diagnostic categories are associated with a higher likelihood of patients leaving hospital against their doctor's advice.


Rosemary Extracts Upregulate Nrf2, Sestrin2, and MRP2 Protein Level in Human Hepatoma HepG2 Cells.
In the past few decades, the incidence of liver cancer has been rapidly rising across the world. Rosemary is known to possess antioxidant activity and is used as natural antioxidant food preservative. It is proposed to have anticancer activity in treating different tumor models. In this study, we try to explore the impact of rosemary extracts on upregulating the level of Nrf2 and Nrf2-regulatory proteins, Sestrin2 and MRP2 in HepG2 cells, and to speculate its potential mechanism. The anticancer activity of rosemary extract, including its polyphenolic diterpenes carnosic acid and carnosol, was evaluated to understand the potential effect on HepG2 cells. Rosemary extract, carnosic acid, and carnosol induced the expression of Sestrin2 and MRP2 associate with enhancement of Nrf2 protein level in HepG2 cells, in which carnosic acid showed most obvious effect. Although the activation pathway of Nrf2/ARE was not exactly assessed, it can be assumed that the enhancement of expression of Sestrin2 and MRP2 may result from upregulation of Nrf2.


Lipid peroxidation products inhibit dopamine catabolism yielding aberrant levels of a reactive intermediate.
Recent work indicates that oxidative stress is a factor in Parkinson's disease (PD); however, it is unknown how this condition causes selective dopaminergic cell death. The neurotransmitter dopamine (DA) has been implicated as an endogenous neurotoxin to explain the selective neurodegeneration. DA undergoes catabolism by monoamine oxidase (MAO) to the reactive intermediate 3,4-dihydroxyphenylacetaldehyde (DOPAL), which is further oxidized to 3,4-dihydroxyphenylacetic (DOPAC) acid via mitochondrial aldehyde dehydrogenase (ALDH). Previous studies found DOPAL to be more toxic than DA, and the major lipid peroxidation products, that is, 4-hydroxynonenal (4HNE) and malondialdehyde (MDA), potently inhibit ALDH. The hypothesis of this work is that lipid peroxidation products inhibit DOPAL oxidation, yielding aberrant levels of the reactive aldehyde intermediate. Treatment of striatal synaptosomes with 2-100 microM 4HNE or 2-50 microM MDA impaired DOPAL oxidation, resulting in elevated [DOPAL]. The aberrant concentration of DOPAL yielded an increase in protein modification by the DA-derived aldehyde, evident via staining of proteins with nitroblue tetrazolium (NBT). Pretreatment of synaptosomes with an MAO inhibitor significantly decreased NBT staining. On the basis of NBT staining, the order of protein reactivity for DA and metabolites was found to be DOPAL>>DOPAC>DA. Mass spectrometric analysis of a model peptide reacted with DOPAL revealed the adduct to be a Schiff base product. In summary, these data demonstrate the sensitivity of DA catabolism to the lipid peroxidation products 4HNE and MDA even at low, physiologic levels and suggest a mechanistic link between oxidative stress and generation of aberrant levels of an endogenous and protein reactive dopaminergic toxin relevant to PD.


Macrofaunal recolonization of copper-contaminated sediments in San Diego Bay.
Effects of Cu-loading on macrofaunal recolonization were examined in Shelter Island Yacht Basin (San Diego Bay, California). Sediments with high and low Cu levels were defaunated and Cu-spiked, translocated, and then placed back into the environment. These demonstrated that the alteration observed in benthic communities associated with Cu contamination occurs during initial recolonization. After a 3-month exposure to sediments with varying Cu levels, two primary colonizing communities were identified: (1) a "mouth assemblage" resembling adjacent background fauna associated with low-Cu levels that was more diverse and predominantly dominated by surface- and subsurface-deposit feeders, burrowers, and tube builders, and (2) a "head assemblage" resembling adjacent background fauna associated with high-Cu concentrations, with few dominant species and an increasing importance of carnivores and mobile epifauna. Cu loading can cause reduced biodiversity and lower structural complexity that may last several months if high concentrations persist, with a direct effect on community functioning.


Antisense c-erbB-2 Decreases DNA Repair and Apoptosis Inducibility in Human Diploid Fibroblasts.
pDOR-erbB-2 sense and antisense retroviral expression vectors were introduced into normal human diploid fibroblasts(2BS), respectively, to construct 2BS-A cells(transfected with antisense recombinant vector) and 2BS-S cells (transfected with sense recombinant vector). Southern blot analysis verified that the exgenous c-erbB-2 cDNA were integrated into genomic DNA. Compared with the cells transfected with pDOR-neo empty vector, 2BS-A cells exhibited decreased ability not only to repair DNA damage, but also to undergo apoptosis. This was consistent with our observations that senescent phenotype was earlier appeared in these cells.


Regulation of B lymphocyte activation by the Fc portion of immunoglobulin.
Murine splenic B lymphocytes are induced to proliferate and undergo polyclonal activation in the presence of Fc fragments, AHGG, antigen-antibody complexes, and CH3 fragments derived from plasmin digestion of human Ig. The unifying feature of the polyclonal antibody response induced by these agents is that in all cases a portion of the constant region of the Ig molecule (ie, Fc region) is present. Fragments of Ig lacking the Fc piece, such as Fab and F(ab)2 were found not to be stimulatory. In addition, a model is proposed to account for the regulatory effects of antigen-antibody complexes on an ongoing humoral immune response.


Embolism to the central nervous system from cardiac myxoma. Report of two cases.
Two cases of left atrial myxoma are reviewed, both presenting as embolic phenomena. Neither patient gave a history compatible with pre-existent cardiac dysfunction. Sudden collapse and subsequent right hemiplegia resulted in one patient when an embolus lodged in the left middle cerebral artery. The second patient presented with headache and transient visual obscuration in the left eye. She showed evidence of embolism to the central retinal artery, and particulate matter could be seen within the retinal arterioles. Attention is drown to the fact that echocardiography now constitutes a simple, noninvasive, and highly reliable method of making this diagnosis. The propensity for embolic tumor fragments to grow and invade cerebral arterial walls is discussed along with its possible neurosurgical significance.


GTL001, a bivalent therapeutic vaccine against human papillomavirus 16 and 18, induces antigen-specific CD8+ T cell responses leading to tumor regression.
Prophylactic vaccines are available for women and girls not yet infected with HPV, but women already infected with HPV need a treatment to prevent progression to high-grade cervical lesions and cancer. GTL001 is a bivalent therapeutic vaccine for eradicating HPV-infected cells that contains HPV16 E7 and HPV18 E7 both fused to detoxified adenylate cyclase from Bordetella pertussis, which binds specifically to CD11b+ antigen-presenting cells. This study examined the ability of therapeutic vaccination with GTL001 adjuvanted with topical imiquimod cream to induce functional HPV16 E7- and HPV18 E7-specific CD8+ T cell responses. Binding of GTL001 to human CD11b was assessed by a cell-based competition binding assay. Cellular immunogenicity of intradermal vaccination with GTL001 was assessed in C57BL/6 mice by enzyme-linked immunospot assay and in vivo killing assays. In vivo efficacy of GTL001 vaccination was investigated in the TC-1 murine HPV16 E7-expressing tumor model. GTL001 bound specifically to the human CD11b/CD18 receptor. GTL001 adjuvanted with topical 5% imiquimod cream induced HPV16 E7 and HPV18 E7-specific CD8+ T cell responses. This CD8+ T-cell response mediated in vivo killing of HPV E7-expressing cells. In the HPV16 E7-expressing tumor model, GTL001 adjuvanted with imiquimod but not imiquimod alone or a combination of unconjugated HPV16 E7 and HPV18 E7 caused complete tumor regression. GTL001 adjuvanted with topical 5% imiquimod is immunogenic and induces HPV16 E7 and HPV18 E7-specific CD8+ T cell responses that can kill HPV E7-expressing cells and eliminate HPV E7-expressing tumors.


Lubiprostone Increases Small Intestinal Smooth Muscle Contractions Through a Prostaglandin E Receptor 1 (EP1)-mediated Pathway.
Lubiprostone, a chloride channel type 2 (ClC-2) activator, was thought to treat constipation by enhancing intestinal secretion. It has been associated with increased intestinal transit and delayed gastric emptying. Structurally similar to prostones with up to 54% prostaglandin E2 activity on prostaglandin E receptor 1 (EP1), lubiprostone may also exert EP1-mediated procontractile effect on intestinal smooth muscles. We investigated lubiprostone's effects on intestinal smooth muscle contractions and pyloric sphincter tone. Isolated murine small intestinal (longitudinal and circular) and pyloric tissues were mounted in organ baths with modified Krebs solution for isometric recording. Basal muscle tension and response to electrical field stimulation (EFS; 2 ms pulses/10 V/6 Hz/30 sec train) were measured with lubiprostone (10(-10)-10(-5) M) ± EP1 antagonist. Significance was established using Student t test and P < 0.05. Lubiprostone had no effect on the basal tension or EFS-induced contractions of longitudinal muscles. With circular muscles, lubiprostone caused a dose-dependent increase in EFS-induced contractions (2.11 ± 0.88 to 4.43 ± 1.38 N/g, P = 0.020) that was inhibited by pretreatment with EP1 antagonist (1.69 ± 0.70 vs. 4.43 ± 1.38 N/g, P = 0.030). Lubiprostone had no effect on circular muscle basal tension, but it induced a dose-dependent increase in pyloric basal tone (1.07 ± 0.01 to 1.97 ± 0.86 fold increase, P < 0.05) that was inhibited by EP1 antagonist. In mice, lubiprostone caused a dose-dependent and EP1-mediated increase in contractility of circular but not longitudinal small intestinal smooth muscles, and in basal tone of the pylorus. These findings suggest another mechanism for lubiprostone's observed clinical effects on gastrointestinal motility.


[Respiratory physiotherapy and respiratory mechanics of chronic respiratory insufficiency].
The effect of short, medium and long term physiotherapy remain controversial. The theoretical objectives of the method are to reduce dyspnoea and respiratory disability while improving exercise performance and expectation of life. As regards respiratory mechanics, the goals are to reduce bronchial obstruction and the mechanical properties of the chest wall. A rise in expiratory flow might result in an increased drainage of secretions, a reduction in transmural bronchial pressure and a raised level of ventilation at end-tidal volume. The correction of parietal distortions by physiotherapy has as an objective a diminution of thoracic elastance. Muscular training, a new aspect of respiratory physiotherapy, consists of increased both the endurance and the force of contraction of the respiratory muscles. The beneficial effects of these recent methods require confirmation with long term clinical studies. The correct indications for the diverse techniques proposed require a better understanding of the mechanism of action of respiratory physiotherapy, an essential complement to effective treatment of chronic airflow obstruction.


Pathologic changes in the liver and kidney produced by immunization with intestinal antigens.
Rabbits were immunized with intestinal antigens derived from rabbits, guinea pigs, and germfree rats. Inflammatory changes throughout the portal tracts of the liver were found in 55% (16 of 29) of the immunized rabbits. Interstitial nephritis was present in 7 of 23 rabbits evaluated. These changes did not occur when nonintestinal antigens were used for immunization. Antigen shared by liver, kidney, duodenum, ileum, and colon were found in each of the species used for immunization. An immune response to the antigen shared by the various tissues may be a factor in the pathogenesis of disease in this experimental system. In man, chronic active hepatitis and interstitial nephritis are found in association with inflammatory bowel disease. A similar mechanism of pathogenesis may be a factor in these extraintestinal manifestations of inflammatory bowel disease in man.


Economic evaluation of famciclovir in reducing the duration of postherpetic neuralgia.
The economic impact of famciclovir therapy for postherpetic neuralgia (PHN) in patients with acute herpes zoster was studied. A decision-analytic model of the treatment of herpes zoster and PHN was used to compare the cost of PHN between patients treated with oral famciclovir 500 mg three times daily for seven days and patients not receiving any antiviral therapy. The effects of famciclovir on PHN in the model were based on the results of a randomized, double-blind trial in 419 adult outpatients. The cost of the course of famciclovir therapy (21 tablets) was estimated as the sum of the drug's wholesale acquisition cost and the pharmacy dispensing cost. The cost of treating PHN (physician visits, medications, and miscellaneous nondrug therapy) was estimated by consulting a panel of physicians. According to the model, the cost of treating PHN was $85 lower per famciclovir recipient ($294 for famciclovir versus $379 for no antiviral therapy). The net cost of famciclovir therapy was $23 per patient ($108 for acquisition and dispensing minus the $85 savings). Among patients 50 years of age or older, famciclovir reduced the average cost of PHN by $155 ($414 for famciclovir versus $569 for no antiviral therapy) and yielded a net savings of $7 per patient. A model for the use of famciclovir to treat acute herpes zoster showed that the cost of such therapy was largely offset by savings in the cost of treating this complication.


Enzyme immunoassay for IgG rheumatoid factor combining with homologous IgG.
An enzyme immunoassay (EIA) was developed to detect, in human sera, IgG rheumatoid factor (RF) combining with human IgG. Wells of the EIA plate were coated with the Fc fragment of human IgG. Binding of RF was detected by goat antibodies to F(ab')2 of human IgG followed by rabbit anti-goat IgG conjugated with alkaline phosphatase and finally by the substrate. Using this procedure, RF was detected in 35%-85% of various pathological human sera, but only in 7% of normal human sera. An analogous procedure was also described for detection, in rabbit sera, of an RF combining with rabbit IgG.


The Emerging Roles of Circular RNAs in the Chemoresistance of Gastrointestinal Cancer.
Gastrointestinal (GI) cancer represents a major global health problem due to its aggressive characteristics and poor prognosis. Despite the progress achieved in the development of treatment regimens, the clinical outcomes and therapeutic responses of patients with GI cancer remain unsatisfactory. Chemoresistance arising throughout the clinical intervention is undoubtedly a critical barrier for the successful treatment of GI cancer. However, the precise mechanisms associated with chemoresistance in GI cancer remain unclear. In the past decade, accumulating evidence has indicated that circular RNAs (circRNAs) play a key role in regulating cancer progression and chemoresistance. Notably, circRNAs function as molecular sponges that sequester microRNAs (miRNAs) and/or proteins, and thus indirectly control the expression of specific genes, which eventually promote or suppress drug resistance in GI cancer. Therefore, circRNAs may represent potential therapeutic targets for overcoming drug resistance in patients with GI cancer. This review comprehensively summarizes the regulatory roles of circRNAs in the development of chemoresistance in different GI cancers, including colorectal cancer, gastric cancer and esophageal cancer, as well as deciphers the underlying mechanisms and key molecules involved. Increasing knowledge of the important functions of circRNAs underlying drug resistance will provide new opportunities for developing efficacious therapeutic strategies against GI cancer.


Biosolids decomposition after surface applications in west Texas.
In a semiarid environment, climate is a critical factor in the decomposition of surface-applied biosolids. This study examined the effect of 2- to 7-yr exposure times on the composition of single applications of New York, NY biosolids in western Texas. Exposure time effects on organic matter, N, P, S, Cu, Cr, Pb, Hg, and Zn were studied near Sierra Blanca, TX. Due to organic matter decomposition, total organic C decreased from 340 g kg(-1) in fresh biosolids to 180 g kg(-1) in biosolids after 82 mo of exposure, whereas the inorganic ash content of the biosolids increased from 339 to 600 g kg(-1). Total N decreased from 50 to 10 g N kg(-1) and total S decreased from 12 to 6 g S kg(-1). Bicarbonate-available P in the biosolids decreased from 0.9 to 0.2 g kg(-1). Successive H2O extractions yielded soluble P concentrations consistent with dicalcium phosphate (dical) for fresh biosolids and tricalcium phosphate (trical) for biosolids exposed for 59 months or more. Sparingly soluble phosphates, such as dical and trical, potentially yield > 0.5 mg P L(-1) in runoff waters for extended periods after biosolids applications, especially after multiple applications. Selective dissolution of the biosolids indicated that as much as 66 to 78% of P exists as iron phosphates, 16 to 21% as Fe oxides, and 5 to 12% as insoluble Ca phosphates. Chemical analyses of ash samples suggest that Cu and Zn have been lost from biosolids through leaching or runoff and no losses of Pb, Cr, or Hg have occurred since application.


Economic costs of anxiety disorders.
Anxiety disorders are estimated to affect 26.9 million individuals in the United States at some point during their lives. This study used the human capital approach to estimate the direct and indirect costs of these highly prevalent disorders. In 1990, costs associated with anxiety disorders were $46.6 billion, 31.5% of total expenditures for mental illness. Less than one-quarter of costs associated with anxiety disorders were for direct medical treatment; over three-quarters were attributable to lost or reduced productivity. Most of these indirect costs were associated with morbidity, as mortality accounted for just 2.7% of the total. Greater availability of effective, relatively low-cost outpatient treatment could substantially reduce the economic and social burden of these common and often crippling disorders.


Development and validation of the Chinese version of dry eye related quality of life scale.
To develop the Chinese version of quality of life scale for dry eye patients based on the Impact of Dry Eye on Everyday Life (IDEEL) questionnaire and to assess the reliability and validity of the developed scale. The original IDEEL was adapted cross-culturally to Chinese language and further developed following standard procedures. A total of 100 Chinese patients diagnosed with dry eye syndrome were included to investigate the psychometric properties of the Chinese version of scale. Psychometric tests included internal consistency (Cronbach's ɑ coefficients), construct validity (exploratory factor analysis), and known-groups validity (the analysis of variance). The Chinese version of Dry Eye Related Quality of Life (CDERQOL) Scale contains 45 items classified into 5 domains. Good to excellent internal consistency reliability was demonstrated for all 5 domains (Cronbach's ɑ coefficients range from 0.716 to 0.913). Construct validity assessment indicated a consistent factorial structure of the CDERQOL scale with hypothesized construct, with the exception of "Dry Eye Symptom-Bother" domain. All domain scores were detected with significant difference across three severity groups of dry eye patients (P < 0.05) except for "Satisfaction with Treatment" domain, indicating good known-groups validity. The results indicated that the CDERQOL scale is a reliable and valid instrument for patients with dry eye syndrome among Chinese population, and could be used as a supplementary diagnostic and treatment-effectiveness measure.


Allosteric P450 mechanisms: multiple binding sites, multiple conformers or both?
According to the initial hypothesis on the mechanisms of cooperativity in drug-metabolizing cytochromes P450, a loose fit of a single substrate molecule in the P450 active site results in a requirement for the binding of multiple ligand molecules for efficient catalysis. Although simultaneous occupancy of the active site by multiple ligands is now well established, there is increasing evidence that the mechanistic basis of cooperativity also involves an important ligand-induced conformational transition. Moreover, recent studies demonstrate that the conformational heterogeneity of the enzyme is stabilized by ligand-dependent interactions of several P450 molecules. Application of the concept of an oligomeric allosteric enzyme to microsomal cytochromes P450 in combination with a general paradigm of multiple ligand occupancy of the active site provides an excellent explanation for complex manifestations of the atypical kinetic behavior of the enzyme.


[Strategy for the treatment of Stage IV gastric cancer with preoperative chemotherapy].
In 2007, we began using neoadjuvant chemotherapy for the treatment of Stage IV gastric cancer and then performing R0/1 surgery in patients in whom chemotherapy was effective. Here, we evaluate the use of this therapeutic strategy combining chemotherapy and surgery for the treatment of Stage IV gastric cancer. The subjects of our investigation were 46 patients with Stage IV gastric cancer treated from 2007 through 2012. We divided these patients into the NAC group (19 patients), in whom we performed R0/1 surgery after chemotherapy, and the Cx group (27 patients), who continued chemotherapy. We also included 79 patients with Stage IV gastric cancer treated from 2001 to 2006, divided into the OPE group (36 patients), in whom we performed R0/1 surgery without neoadjuvant chemotherapy, and the NC group (43 patients), in whom we performed R2 surgery. We plotted the survival curves of these 4 groups. The chemotherapy protocols used were S-1+cisplatin( CDDP) and S-1+docetaxel( DOC). The disease control rate of these chemotherapies was 72%, and R0/1 surgery was performed in 53.8% of patients with liver metastasis, 62.5% of those with paraaortic lymph node (PALN) metastasis, 29.4% of those with peritoneal metastasis, 100% of patients with T4N2 disease, and 0% of patients with distant metastasis. The 2-year survival rates of the NAC, OPE, Cx, and NC groups were 69%, 55%, 0%, and 20%, respectively. The 5-year survival rates of the NAC, OPE, Cx, and NC groups were 35%, 30%, 0%, and 5%, respectively.


Zooming into Focus Groups: Strategies for Qualitative Research in the Era of Social Distancing.
Qualitative research methods are important and have become increasingly prominent in medical education and research. The reason is simple: many pressing questions in these fields require qualitative approaches to elicit nuanced insights and additional meaning beyond standard quantitative measurements in surveys or observatons. Among the most common qualitative data collection methods are structured or semistructured in-person interviews and focus groups, in which participants describe their experiences relevant to the research question at hand. In the era of physical and social distancing because of the novel coronavirus disease (COVID-19) pandemic, little guidance exists for strategies for conducting focus groups or semistructured interviews. Here we describe our experience with, and recommendations for, conducting remote focus groups and/or interviews in the era of social distancing. Specifically, we discuss best practice recommendations for researchers using video teleconferencing programs to continue qualitative research during the COVID-19 pandemic.


Alcohol policy and the public good.
Despite a diversity of cultural, social and economic experiences with alcohol at national level, a broad unity of purpose is emerging within the alcohol problems perspective. The scientific basis for that perspective finds its support in a sustained international research effort. With the publication in 1994 of the WHO report "Alcohol Policy and the Public Good", that perspective has been given persuasive empirical underpinning and a sharp focus. Essentially the problems perspective suggests that the target for public policy is the prevention or alleviation of population-wide alcohol-related problems, with problems broadly defined. The test for any policy within this arena is then whether it contributes to the achievement of that target. The perspective is thus rational, empirical, rooted in the traditions of public health, and readily intelligible to the ordinary citizen. Key conclusions emanating from "Alcohol Policy and the Public Good" are summarized. That text is rounded off by the challenging assertion that "Drinking problems are not carried by an uncontrollable tide. With public will, and on the evidence, they are capable of amelioration." A positive response to that challenge by the nations of Europe is both necessary and feasible.


Molecular Information Theory Meets Protein Folding.
We propose an application of molecular information theory to analyze the folding of single domain proteins. We analyze results from various areas of protein science, such as sequence-based potentials, reduced amino acid alphabets, backbone configurational entropy, secondary structure content, residue burial layers, and mutational studies of protein stability changes. We found that the average information contained in the sequences of evolved proteins is very close to the average information needed to specify a fold ∼2.2 ± 0.3 bits/(site·operation). The effective alphabet size in evolved proteins equals the effective number of conformations of a residue in the compact unfolded state at around 5. We calculated an energy-to-information conversion efficiency upon folding of around 50%, lower than the theoretical limit of 70%, but much higher than human-built macroscopic machines. We propose a simple mapping between molecular information theory and energy landscape theory and explore the connections between sequence evolution, configurational entropy, and the energetics of protein folding.


Paraganglion of the prostate in a needle biopsy: a potential diagnostic pitfall.
Paraganglionic tissue incidentally observed in a needle biopsy of the prostate is reported. The tissue was seen in the periprostatic adipose tissue obtained during a needle biopsy of the prostate of an 81-year-old man. The paraganglionic cells demonstrated round to oval nuclei and basophilic granular or vacuolated cytoplasm and were immunohistochemically positive for chromogranin A, but negative for prostatic acid phosphatase and prostate-specific antigen. Paraganglion in the periprostatic adipose tissue is a diagnostic pitfall and should be distinguished from extension of prostatic adenocarcinoma outside the prostatic capsule into the periprostatic adipose tissue.


Improved photostability indicating ion-pair chromatography method for pergolide analysis in tablets and in the presence of cyclodextrins.
Pergolide (PG) a semi-synthetic ergot alkaloid derivative used mainly for the treatment of Parkinson's disease is known to be a photosensitive drug substance. The major photodegradation products are PG sulphoxide (SX) and PG sulphone (SN), which are also the main impurities of the bulk drug substance. It is widely metabolized to more than 10 metabolites including SX and SN. In this work an improved photostability indicating ion-pair chromatography method for PG mesilate was developed. The method can be applied in the determination of PG and impurities in aqueous solutions and in tablets for routine analysis. This new method is appropriate for the quantitative determination of PG in the presence of its impurities and photodegradation products and can also be used for PG complexes with cyclodextrins (commonly used as photostabilizing agents). Furthermore it is suitable for the quantitation of its impurities and its thermal or photo-induced decomposition products. Separation was achieved on a ThermoQuest C(18) BDS column and Sodium octanosulphonate was used as ion-pairing agent. Analysis was performed at 223 nm. Validation parameters included: specificity, linearity, precision and accuracy, limit of quantitation and suitability. The method was found to be specific and linear for PG, as well as for SX and, SN impurities. The recovery was 100.83+/-0.46% for PG, 99.86+/-0.33% for SX and 99.77+/-1.84% for SN. Finally the photodegradation profile of PG mesilate was studied in different initial sample concentration. The obtained result revealed that: PG photolysis is catalyzed by its degradation products and that decrease of initial sample concentration reduces the rate of PG photoinduced degradation.


Nanoneedle-Based Sensing in Biological Systems.
Nanoneedles are high aspect ratio nanostructures with a unique biointerface. Thanks to their peculiar yet poorly understood interaction with cells, they very effectively sense intracellular conditions, typically with lower toxicity and perturbation than traditionally available probes. Through long-term, reversible interfacing with cells, nanoneedles can monitor biological functions over the course of several days. Their nanoscale dimension and the assembly into large-scale, ordered, dense arrays enable monitoring the functions of large cell populations, to provide functional maps with submicron spatial resolution. Intracellularly, they sense electrical activity of complex excitable networks, as well as concentration, function, and interaction of biomolecules in situ, while extracellularly they can measure the forces exerted by cells with piconewton detection limits, or efficiently sort rare cells based on their membrane receptors. Nanoneedles can investigate the function of many biological systems, ranging from cells, to biological fluids, to tissues and living organisms. This review examines the devices, strategies, and workflows developed to use nanoneedles for sensing in biological systems.


A novel spliced variant of the TIN2 shelterin is present in chronic lymphocytic leukemia.
The shelterin proteins play important roles in telomere maintenance and genome stability. These proteins have been found to be mutated in many cancers including CLL. Herein, we demonstrate here the presence of a novel spliced isoform of TIN2S in chronic lymphocytic leukemia (CLL), related to deletion of exon 2 in the TIN2 gene. The expressions of spliced TIN2S mRNA varied widely in CLL and there was an inverse relationship between the mRNA levels of full-length TIN2S and the spliced moiety. Small amounts of spliced TIN2S were also observed in normal B cells but not in T cells. Spliced TIN2S appeared dysfunctional, as immunoprecipitation studies showed the typical association of TRF2 and TIN2 in normal lymphocytes but not in CLL cells. Moreover, whereas TRF2 localized to the nucleus in normal lymphocytes, it was present in both nuclei and cytoplasm in CLL cells. The levels of spliced TIN2S increased with age and in 3 of 8 patients increased over time. The presence of the spliced variant failed to be related to telomere length in CLL suggesting other functions for this protein. Further studies are required to determine the etiology and biological significance of this unique spliced TIN2S variant.


Evaluation of a preprimed microporous hollow-fiber membrane for rapid response neonatal extracorporeal membrane oxygenation.
Delays in initiating extracorporeal membrane oxygenation (ECMO) in the critically ill pediatric patient may lead to adverse outcomes. Maintaining a primed ECMO circuit can considerably reduce the initiation time. The predominant concerns precluding this practice are a decrease in oxygenator efficiency due to the saturation of microporous hollow fibers and compromised sterility when the oxygenator has been primed for 30 days. For institutions using a hollow-fiber oxygenator for ECMO, there are no data reporting pre-primed hollow-fiber oxygenator viability. This study reports the efficiency of oxygen transfer and the sterility of the Carmeda Minimax Plus (Medtronic, Inc, Minneapolis, MN) oxygenator after being crystalloid primed for 30 days. A total of 10 Minimax Plus oxygenators were tested for oxygen transfer in a laboratory setting utilizing fresh whole bovine blood. The control group (n =5) were tested immediately after priming. The test group (n =5) were oxygenators primed for 30 days with crystalloid solution and left stagnant until tested. Prior to testing, all oxygenators were circulated for 5 min and samples drawn to test for circuit sterility. Venous inlet saturations were manipulated to achieve three levels of testing: venous saturation (SvO2) of 55% for an oxygen challenge, SvO2 of 65% to comply with AAMI standards, and SvO2 of 75% to assess oxygen transfer rates and peak PaO2 achievement. Blood flow for all tests was maintained at 2 L/min with 1:1 blood to gas flow ratio and 100% FiO2. Samples were drawn pre- and postoxygenator at 1- and 6-hour time intervals to compute actual oxygen transfer values. All cultures from the test group priming solution produced no microbial growth after 30 days of stagnant prime. Average oxygen transfer values (ml/O2/min) for the control group after 1 hour of continuous use were 130.1+/-15.5 (@ 55% SvO2), 113.7+/-10.4 (@ 65% SvO2), 97.7+/-8.9 (@ 75% SvO2). After 6 hours, the average transfer values increased to 134.2+/-13.2 (@ 55% SvO2), 118.76+/-6.6 (@ 65% SvO2) and 98.9+/-8.3 (@ 75% SvO2). The average oxygen transfer values after 1 hour for oxygenators primed for 30 days were 114.9+/-10.0 (@ 55% SvO2), 112.4+/-8.2 (@ 65% SvO2) and 89.6+/-16.0 (@ 75% SvO2). After 6 hours of use, the average transfer values all decreased to 111.4+/-2.1 (@ 55% SvO2, p <0.05 versus control), 104.0+/-5.6 (@ 65% SvO2, p <0.05 versus control) and 88.4+/-3.2 (@ 75% SvO2, p <0.05 versus control). In conclusion, there was a decrease in the average oxygen transfer values for the test group after 6 hours versus the control. The modest loss of oxygen transfer ability observed can be considered acceptable due to the amount of surface area of the Minimax Plus oxygenator when used on a neonate, making it feasible to adopt the practice of prepriming the Minimax oxygenator for neonatal ECMO.


Catalytic N-Si coupling as a vehicle for silane dehydrocoupling via α-silylene elimination.
Exploration of (N3N)ZrNMe2 (1, N3N = N(CH2CH2NSiMe3)33-) as a catalyst for the cross-dehydrocoupling or heterodehydrocoupling of silanes and amines suggested silylene reactivity. Further studies of the catalysis and stoichiometric modeling reactions hint at α-silylene elimination as the pivotal mechanistic step, which expands the 3p elements known to engage in this catalysis and provides a new strategy for the catalytic generation of low-valent fragments.


Dorsal Subluxation of The Proximal Carpal Row with the Use of a Bridge Plate.
Background Spanning bridge plates were first popularized for fixation of complex distal radius fractures. However, indications for their use have expanded including the surgical treatment algorithm for treating conditions such as Kienböck's disease. Traditionally, initial surgical treatment of Lichtman Stages II to III Kienböck's disease included lunate decompression, unloading, and revascularization procedures. The addition of a dorsal spanning bridge plate further facilitates lunate offloading and may improve bone revascularization. Case Description We report a complication of proximal carpal row dorsal subluxation secondary to dorsal spanning plate fixation in a patient with Stage IIIb Kienböck's disease. The patient had undergone wrist arthroscopy, lunate forage, radius core decompression, and spanning plate fixation. At 6 weeks after surgery, radiographic imaging demonstrated dorsal subluxation of the proximal carpal row that was corrected upon bridge plate removal. Serial radiographs during follow-up showed no further carpal subluxation without Kienböck's disease progression at 12 months postremoval of hardware. Patient remains pain free and has returned to elite level sport. Literature Review To our knowledge no previous cases of proximal carpal row subluxation with the use of dorsal bridge plate has been reported in the literature. Clinical Relevance Proximal row carpal subluxation can occur with dorsal bridge plate fixation.


Intra-scrotal extra-testicular schwannoma: A case report and literature review.
Intra-scrotal schwannoma is a rare neoplasm and a few reports were describing this entity in the literature and mostly difficult to be diagnosed pre operatively(1) We recently treated a case of intra-scrotal extra-testicular schwannoma which was discovered in a patient with history of painless scrotal lesion for 5 years. paratesticular lesion excision was done which was result as schwannoma tissue. follow up with US scrotum was unremarkable for the patient. surgical excision will provide diagnostic and therapeutic goals. Even tough recurrence is rare a urologist should take care to ensure complete surgical resection.


Control of proliferative enteropathy in growing/fattening pigs using growth promoters.
The aim of this study was to evaluate the effect of different antibiotics used as growth promoters on the control of porcine intestinal adenomatosis when administered in weaning, growing and fattening pig diets, according to Annex I of the European Union directive (70/524/EEC and its subsequent amendments to date) for the use of feed additives. On a farm with a previous history of proliferative enteropathy outbreaks, 648 weaned piglets (23 days old) were divided into nine experimental groups according to bodyweight and sex ratio, each group comprising four pens with 18 pigs in each pen. One group served the trial as a negative (unmedicated) control: another (the positive control) received monensin via feed at 100 p.p.m. up to the end of the growing phase (107 days old) and 50 p.p.m. up to slaughter age (156 days old). The remaining seven groups were offered feed with the addition of the following antibiotics: virginia-mycin (50-20 p.p.m.), avilamycin (40-20 p.p.m.), spiramycin (50-20 p.p.m.), zinc bacitracin (50-10 p.p.m.), avoparcin (40-20 p.p.m.), tylosin (40-20 p.p.m.) and salinomycin (60-30 p.p.m.), respectively. The performance of the pigs in the positive control group was very satisfying and among the highest in the trial, verifying earlier field studies. As a general conclusion it seems that all tested growth promoters had a beneficial effect compared with the untreated control, indicated by the decrease of mortality rate, the elimination of diarrhoeal incidence and the enhancement of growth performance, although the proliferative enteropathy control achieved by each substance was not always satisfactory. More specifically, the antibiotic growth promoters tested can be scaled according to their total efficacy as follows: 1. Salinomycin, tylosin, spiramycin; 2. Virginiamycin, zinc bacitracin, avilamycin; and 3. Avoparcin. Finally, it is considered that part of the growth promotion efficacy of the tested substances is due to their potential capacity to control porcine intestinal adenomatosis; thus, in future growth performance trials, the disease background of the trial farms must be examined, especially for porcine enteropathy challenges.


Advances in modeling and treating spinal muscular atrophy.
Spinal muscular atrophy (SMA) is an inherited childhood neurodegenerative disorder caused by ubiquitous deficiency of the survival motor neuron (SMN) protein - the hallmarks of which are the selective loss of motor neurons and skeletal muscle atrophy. Here, we highlight recent progress in the understanding of SMA pathology and in the development of therapeutic approaches for its treatment. Phenotypic characterization of mouse models of the disease, combined with analysis of SMN restoration or depletion in a spatially and temporally controlled manner, has yielded key insights into the normal requirement of SMN and SMA pathophysiology. Increasing evidence indicates a higher demand for SMN during neuromuscular development and extends the pathogenic effects of SMN deficiency beyond motor neurons to include additional cells both within and outside the nervous system. These findings have been paralleled by preclinical development of powerful approaches for increasing SMN expression through gene therapy or splicing modulation that are now in human trials. Along with the availability of SMN-upregulating drugs, identification of the specific cell types in which SMN deficiency induces the disease and delineation of the window of opportunity for effective treatment are key advances in the ongoing path to SMA therapy.


The formation of iron complexes with bile and bile constituents.
Inorganic iron is able to form complexes with whole bile and with some bile constituents and these remain soluble at neutral pH. Ascorbic acid does not appear to play a part in this process. The formation of soluble complexes in vivo is an important factor in maintaining intraluminal iron in a form available for absorption, and bile may have some importance in this respect.


Synthesis, 5-HT1A and 5-HT2A receptor activity of new 1-phenylpiperazinylpropyl derivatives with arylalkyl substituents in position 7 of purine-2,6-dione.
A series of new 1,3-dimethyl-7-phenylalkyl-8-[3-(4-phenyl-1-piperazinyl)propylamino]-purine-2,6-dione derivatives (10-16) was synthesized and their 5-HTIA and 5-HT2A receptor affinities were determined. It was found that compounds with the phenylpropyl substituent in position 7 of purine-2,6-dione (12, 14 and 16), or with phenylmethyl in position 7 and 2-OCH3 in the phenylpiperazine part (13) showed a distinct affinity for 5-HTIA receptors (Ki = 8-50 nM). No structural modifications resulted in 5-HT2A ligands, since the affinity of 10-16 for those receptors was insignificant (Ki = 115-550 nM). The new 5-HT1A receptor ligands (12-14, 16) were investigated in vivo to determine their functional activity at those receptors. In behavioral studies, 12-14 and 16 behaved like postsynaptic 5-HTIA receptor antagonists, since they reduced lower lip retraction and the behavioral syndrome induced by 8-OH-DPAT (5-HT1A receptor agonist) in rats. When given alone, none of the compounds investigated in vivo, mimicked 8-OH-DPAT activity in those tests. Derivative 12 did not affect the body temperature in mice, whereas 13, 14 and 16 decreased it. Furthermore, 12 did not change the hypothermia induced by 8-OH-DPAT, and the decrease in body temperature in mice induced by 13, 14 or 16 was not antagonized by WAY 100635 (5-HT1A receptor antagonist); hence in that model neither 12, 13, 14 nor 16 acted as antagonists or agonists, respectively, at presynaptic 5-HT1A receptors.


Leflunomide: an immunosuppressive drug with multiple effects on T cell function.
Abstract Leflunomide has recently been introduced as a new treatment for rheumatoid arthritis. Although its immunosuppressive effect has been well demonstrated in experimentally induced autoimmune diseases and in organ transplant rejection in animal models, the exact mechanisms mediating its immunomodulatory effect are not fully understood. As T cells play a central role in the orchestration of immune responses in both physiological and pathological conditions, it has been proposed that the ability of leflunomide to suppress inappropriate and unwanted immunity is related to a functional inhibition of T cells. A precise knowledge of the mechanisms of leflunomide's action on T cells is therefore necessary. As the clinical effect of leflunomide has been well described elsewhere, this review will focus on, and will discuss, current data on the different aspects of leflunomide's effect on T cell function.


Study of starch aging characteristics based on Terahertz technology.
Traditional methods for the determination of starch aging indicators often have a series of shortcomings such as time-consuming, high cost, large human error, damage to samples, environmental pollution, and high requirements for inspectors. Therefore, it is meaningful to find or establish a dynamic fingerprint identification pattern that can detect the aging degree of starch during the process of processing or storage quickly and accurately. It not only provides guidance for starch food processing but also saves a lot of human, material resources, and time. Terahertz technology is an emerging molecular spectroscopy technology in the 21st century. It is with low energy and basically harmless to the human body. It can also realize nondestructive testing of samples. In the experiment, the samples were prepared by the tableting method and the samples containing 20% of 50 mg samples were prepared with polyethylene as the diluent. The thickness of the samples was 1 mm and the diameter was 13 mm. The terahertz time-domain spectrometer was used to obtain the spectral information of aging starch at different aging times. After the pretreatment of the spectrum by vector normalization, first derivative, and multiple scattering correction, the prediction models of aging days, crystallinity, and resilience of aging starch were established, respectively. The determination coefficient (R 2) of the established models is all greater than 95%, indicating that the established models are highly reliable and can be used to predict the aging days, crystallinity, and retrogradation degree of starch. And the R 2 of the prediction model based on the refractive index spectrum is greater than that of the absorption coefficient spectrum. The experimental method obtains the dynamic fingerprint identification map of starch in the aging process, realizes the real-time monitoring and detection of the starch aging process, and provides an effective means for the production and processing of starch-related industries.


Fracture Resistance and Fracture Behaviour of Monolithic Multi-Layered Translucent Zirconia Fixed Dental Prostheses with Different Placing Strategies of Connector: An in vitro Study.
To evaluate the effect of different placing strategies performed in the connector area on fracture resistance and fracture behaviour of monolithic multi-layered translucent zirconia fixed dental prostheses (FDPs). Thirty 3-unit monolithic FDPs were produced and divided into three groups (n = 10) based on the different strategies for placing the connector area of FDPs in multi-layered zirconia blank with varying contents of yttria ranging from 4 to 5 mol%. The groups were as follows: FDPs with connectors placed in dentin layer with 4 mol% yttria content, FDPs with connectors placed in gradient layer, and FDPs with connectors placed in translucent layer with 5 mol% yttria content. A final group (n = 10) of conventional monolithic zirconia with a monolayer of yttria content (4 mol%) has been used as a control group. The specimens were artificially aged using thermocycling and pre-loading procedures and subsequently loaded to fracture using a universal testing machine. Fracture loads and fracture behaviour were analyzed using one-way ANOVA and Fisher's exact tests and statistically evaluated (p ≤ 0.05). There were no significant differences in fracture loads among the groups based on the placing strategies of the connector area of the FDPs in the multi-layered translucent zirconia blank (p > 0.05). There was no significant difference in fracture loads between monolithic multi-layered translucent zirconia and conventional monolithic translucent zirconia materials (p > 0.05). Fracture behaviour of FDPs with connector area placed in translucent layer differed significantly compared to FDPs with connector area placed in dentin layer and FDPs in control group (p = 0.004). The placing strategies of the connector used in the computer aided design and manufacturing procedures do not considerably affect fracture resistance of monolithic FDPs made of multi-layered translucent zirconia. Monolithic FDPs made of multi-layered translucent zirconia show comparable strength to FDPs made of conventional translucent zirconia, but with different fracture behaviour.


Monoubiquitination and cellular distribution of XIAP in neurons after traumatic brain injury.
XIAP is a member of the inhibitor of apoptosis (IAP) gene family that, in addition to suppressing cell death by inhibition and polyubiquitination of caspases, is involved in an increasing number of signaling cascades. Moreover, the function and regulation of XIAP in the central nervous system (CNS) is poorly understood. In this study, the authors investigated the cell-type expression, the subcellular distribution, ubiquitination of XIAP, and levels of Smac/DIABLO in the normal adult rat brain and in brains subjected to moderate traumatic brain injury (TBI). In the normal brain, XIAP was predominantly expressed in the perinuclear region of neurons. Traumatized brains showed dramatic alterations in cellular and regional expression of XIAP early after injury. Stereologic analyses of the number of XIAP-positive cells within the hippocampus of both hemispheres showed a biphasic response. Immunoprecipitation and immunoblots of extracts derived from different brain regions demonstrated that a single ubiquitin modifies XIAP. Normal cortex contained significantly higher levels of monoubiquitinated XIAP than hippocampus. TBI induced alterations in levels of monoubiquitinated XIAP that correlated with changes in XIAP distribution and immunoreactivity, suggesting that monoubiquitination of XIAP may be a regulator of XIAP location or activity. Similar levels of Smac/DIABLO were present in lysates of normal and traumatized brains. These data demonstrate for the first time a region-specific regulation of XIAP monoubiquitination in the normal adult rat brain, and after TBI, that may be a key event in the regulation of XIAP function contributing to the pathogenesis following injury.


Enrichment of peptides containing consensus sequence by an enzymatic approach for targeted analysis of proteins.
Multiple residues with consensus sequence, i.e. motif, on proteins are closely related to protein function. However, there is no effective method for targeted analysis of such proteins. The challenge for analysis of these classes of proteins by MS is how to selectively enrich peptides containing consensus sequence from protein digest. Although enrichment of peptides containing one type of amino acid residue was successfully achieved by chemically labeling followed by chromatographic isolation, however, it is almost impossible to label and isolate signature peptides containing multiple residues with consensus sequence by chemical approach. Herein, we developed an enzymatic approach based on the specific recognition between enzyme and its substrates to enrich such peptides. This approach was realized by modification of a residue in the consensus sequence via enzyme that can recognize the sequence followed by the isolation of the modified peptides. cAMP-dependent protein kinase was used to validate this approach and 168 peptides containing consensus motif were identified with selectivity of 67.2%. Those peptides resulted in the identification of 88 proteins with consensus sequence from serum sample. As this motif-oriented peptide enrichment approach allows targeted analysis of a subset of proteins with consensus sequence, it will have broad application in biological studies.


Effects of different gastrointestinal surgical approaches on pancreatic β-cell function in type 2 diabetic patients with BMIs<30.
Gastrointestinal surgery has been performed for many years to achieve durable and effective treatments of type 2 diabetes mellitus (T2DM). This study determined the effects of different surgical operations on pancreatic β-cell function in patients with T2DM with BMIs<30 kg/m(2). A total of 46 patients with diabetes and BMIs<30 kg/m(2) underwent gastrointestinal surgery; 23 patients underwent Roux-en-Y gastric bypass (RYGB), and 23 patients underwent Billroth Ι gastrectomy (BΙG). Oral glucose tolerance tests were used to evaluate pancreatic β cell function. Serum insulin was measured by enzyme-linked immunosorbent assay (ELISA), and blood glucose was detected with the glucose oxidase method. HOMA-IR and HOMA-β were evaluated with the HOMA equation. The T2DM remission rates were 21.7% (5/23) in the BΙG group and 82.6% (19/23) in the RYGB group. There was a significant difference in the improvements in postoperative glycosylated hemoglobin A1c (GHbA1c) and glycated hemoglobin A1 (GHbA1) between the BΙG and RYGB groups (P=0.001, P=0.002). OGTT revealed that both fasting blood glucose (FBG) and blood glucose at the designated time points after glucose loading were significantly lower in the RYGB group than in the BΙG group. Insulin levels (at 30 and 60 min), insulin release indices (at 30 and 60 min), HOMA-β and ΔI30/ΔG30 were significantly higher in the RYGB group than in the BΙG group (P<0.05). Patients with BMIs<30 kg/m(2) who underwent RYGB surgery exhibited significantly greater improvements or resolutions of T2DM and significantly better recoveries of β-cells function than did those in the BΙG group.


Ensaculin (KA-672 HCl): a multitransmitter approach to dementia treatment.
Ensaculin, a novel benzopyranone substituted with a piperazine moiety, showed memory-enhancing effects in paradigms of passive and conditioned avoidance in both normal and artificially amnesic rodents. It exhibited neuroprotective activities in an NMDA toxicity model and neurotrophic effects in primary cultured rat brain cells. The compound could be characterized as a weak NMDA receptor-operated channel blocker. In receptor-binding studies, ensaculin was found to have high affinities to serotonergic 5-HT(1A) and 5-HT(7) receptors, adrenergic alpha(1), and dopaminergic D(2) and D(3) receptors. Due to its unique pharmacodynamic profile, ensaculin may have potential as an antidementia agent acting on various transmitter systems.


Effect of mandibular nerve block on postoperative analgesia in patients undergoing oropharyngeal carcinoma surgery under general anaesthesia.
Postoperative analgesia after oropharyngeal carcinoma surgery remains poorly studied. This study investigates the effects of mandibular nerve block (MNB) with ropivacaine 10 mg ml(-1) in conjunction with general anaesthesia (GA) on postoperative analgesia after partial glossectomy or transmandibular lateral pharyngectomy. In a randomized double-blind study, 42 patients (21 in each group) received an MNB by the lateral extra-oral approach (MNB group) or a deep s.c. injection of normal saline (control group). Both groups received a standardized general anaesthetic. Postoperative analgesia included fixed dose of i.v. acetaminophen and morphine via a patient-controlled analgesia device. Consumption of morphine and supplemental analgesics and pain scores at rest were measured. The mean cumulative morphine consumption was reduced by 56 and 45% at 12 and 24 h after operation in the MNB group. The administration of analgesic rescue medications was delayed in the MNB group. The visual analogue scale (VAS) pain scores were comparable in the two groups during the first 24 h. Adequate analgesia (mean VAS < or = 3) was observed throughout the study period in the MNB group, but only from 4 h after operation onwards in the control group. The number of patients who experienced severe pain (VAS > 7) during the first postoperative day was lower in the MNB group than in the control group (3 vs 10. respectively, P < 0.05). In this study, MNB performed before GA for oropharyngeal carcinoma surgery improved postoperative analgesia, resulting in reduced morphine consumption at 24 h and severe pain in fewer patients.


Analysis of incomplete longitudinal binary data using multiple imputation.
We propose a propensity score-based multiple imputation (MI) method to tackle incomplete missing data resulting from drop-outs and/or intermittent skipped visits in longitudinal clinical trials with binary responses. The estimation and inferential properties of the proposed method are contrasted via simulation with those of the commonly used complete-case (CC) and generalized estimating equations (GEE) methods. Three key results are noted. First, if data are missing completely at random, MI can be notably more efficient than the CC and GEE methods. Second, with small samples, GEE often fails due to 'convergence problems', but MI is free of that problem. Finally, if the data are missing at random, while the CC and GEE methods yield results with moderate to large bias, MI generally yields results with negligible bias. A numerical example with real data is provided for illustration.


Smoking cessation: barriers to success and readiness to change.
Smoking cessation interventions should be individualized based on patient history and readiness for change. The objective of this study was to assess stages of change and key components of smoking and cessation history among a sample of primary care patients. A telephone survey of current or recent smokers identified smoking status, stage of change, motivation, concerns, relapse history, pharmacotherapy, and social support. Of 150 participants, most were within precontemplation (22.7 percent) or contemplation (44.0 percent) stages of change; 14.0 percent were in preparation, 4.7 percent in action, and 14.7 percent in maintenance. The primary motivation for quitting was to improve general health (42.3 percent). The most common cessation-related concerns were: breaking the habit, stress, and weight gain. Pharmacotherapy was discontinued due to adverse events in 31.5 percent of users. Intratreatment social support was reported by 17.5 percent. The most common reasons for relapse were falling back into the habit (36 percent), stressful situations (27 percent), and being around other smokers (25 percent). Targeted interventions are needed for patients in either precontemplation or contemplation stages. Counseling should focus on helping patients resolve barriers to cessation and reasons for relapse, particularly stress and weight management. Pharmacotherapy should be utilized when patients are ready to quit. Increased intratreatment social support and counseling appear warranted to support behavior change and appropriate medication use.


Genetic variants in ANCA-associated vasculitis: a meta-analysis.
Genetic factors may influence the pathogenic pathways leading to antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). We performed a meta-analysis to determine the genetic variants most likely associated with AAV and investigated whether diagnostic and serological subtypes within AAV have distinct genetic backgrounds. Studies investigating the association between genetic variants and AAV in humans were searched in PubMed, EMBASE and Web of Science. All variants investigated in at least two studies were selected. Subsequently, all studies assessing these variants were included in this meta-analysis. Additionally, data on these variants from the largest genome-wide association studies in AAV were included to increase the validity of this meta-analysis. The literature search yielded 5180 articles. 62 articles investigating 140 genetic variants were included, 33 of which were associated with AAV in a meta-analysis. These genetic variants were in or near the following genes: CD226, CTLA-4, FCGR2A, HLA-B, HLA-DP, HLA-DQ, HLA-DR, HSD17B8, IRF5, PTPN22, RING1/RXRB, RXRB, STAT4, SERPINA1 and TLR9. Moreover, we identified genetic distinctions between granulomatosis with polyangiitis and microscopic polyangiitis and between proteinase 3 ANCA vasculitis and myeloperoxidase ANCA vasculitis. In 76% of the genetic variants, subdivision based on ANCA serotype resulted in higher ORs than subdivision based on clinical diagnosis. This meta-analysis identified 33 genetic variants associated with AAV, supporting a role for alpha-1-antitrypsin, the major histocompatibility complex system, and several distinct inflammatory processes in AAV pathogenesis. Our results indicate that subdivision of AAV based on ANCA serotype has a stronger genetic basis than subdivision based on clinical diagnosis.