Sulfonamide as amide isostere for fine-tuning the gelation properties of physical gels.
(S)-2-Stearamidopentanedioic acid (C18-Glu) is a known LMW gelator that forms supramolecular gels in a variety of solvents. In this work, we have carried out the isosteric substitution of the amide group by a sulfonamide moiety yielding the new isosteric gelator (S)-2-(octadecylsulfonamido)pentanedioic acid (Sulfo-Glu). The gelation ability and the key properties of the corresponding gels were compared in terms of gelation concentration, gel-to-sol transition temperature, mechanical properties, morphology, and gelation kinetics in several organic solvents and water. This comparison was also extended to (S)-2-(4-hexadecyl-1H-1,2,3-triazol-4-yl)pentanedioic acid (Click-Glu), which also constitutes an isostere of C18-Glu. The stabilizing interactions were explored through computational calculations. In general, Sulfo-Glu enabled the formation of non-toxic gels at lower concentrations, faster, and with higher thermal-mechanical stabilities than those obtained with the other isosteres in most solvents. Furthermore, the amide-sulfonamide isosteric substitution also influenced the morphology of the gel networks as well as the release rate of an embedded antibiotic (vancomycin) leading to antibacterial activity in vitro against Staphylococcus aureus.


Practical and highly selective sulfur ylide-mediated asymmetric epoxidations and aziridinations using a cheap and readily available chiral sulfide: extensive studies to map out scope, limitations, and rationalization of diastereo- and enantioselectivities.
The chiral sulfide, isothiocineole, has been synthesized in one step from elemental sulfur, γ-terpinene, and limonene in 61% yield. A mechanism involving radical intermediates for this reaction is proposed based on experimental evidence. The application of isothiocineole to the asymmetric epoxidation of aldehydes and the aziridination of imines is described. Excellent enantioselectivities and diastereoselectivities have been obtained over a wide range of aromatic, aliphatic, and α,β-unsaturated aldehydes using simple protocols. In aziridinations, excellent enantioselectivities and good diastereoselectivities were obtained for a wide range of imines. Mechanistic models have been put forward to rationalize the high selectivities observed, which should enable the sulfide to be used with confidence in synthesis. In epoxidations, the degree of reversibility in betaine formation dominates both the diastereoselectivity and the enantioselectivity. Appropriate tuning of reaction conditions based on understanding the reaction mechanism enables high selectivities to be obtained in most cases. In aziridinations, betaine formation is nonreversible with semistabilized ylides and diastereoselectivities are determined in the betaine forming step and are more variable as a result.


What constitutes the best sex life for gay and bisexual men? Implications for HIV prevention.
While a large body of research has sought to understand HIV transmission risk behaviours among gay men, bisexual men and other men who have sex with men (MSM), less attention has been paid to the wider sexual health and well-being of this population. While some community-based organisations aim to support a more holistic sense of sexual well-being there is little evidence to draw on to inform their interventions. The current study sought to explore gay and bisexual men's conceptions of what constitutes the 'best sex'. The EMIS survey of 2010 recruited more than 180,000 respondents from 38 European countries to complete an online questionnaire about sexual health and behaviour. The 12,942 English language, UK-based responses to the open ended question, "What's your idea of the best sex life?" were subjected to a detailed content analysis. A framework was devised to reflect and describe the key themes emerging from the data, which was then used to code all responses to one (or more) of these themes. Further statistical analysis sought to establish if and how responses differed according to key demographic variables. Eight themes emerged that capture the diversity of gay and bisexual men's sexual desires. Most common among responses was a desire for sex within committed relationships, followed by a desire for sex which is emotionally or psychologically connected. Men also expressed a desire for volume and variety in their sexual lives, and for sex that is free from physical, social or psychological harm. Comparative analysis identified that older men were less likely to idealise a relationship or emotional connection, but were more likely to specify the sexual acts or behaviours they wished to engage in. Attending to what men value or aspire to can help ensure interventions are engaging and meaningful to the target population. HIV prevention interventions need to attend to the broad range of sexual desires held by gay and bisexual men in delivery of holistic sexual health promotion that can help them to have the best sex with the least harm.


Malignant breast epithelium selects for insulin-like growth factor II expression in breast stroma: evidence for paracrine function.
Paracrine interactions between stromal and epithelial cells are important influences on the growth and malignant behavior of breast cancers. Insulin-like growth factors I and II (IGF-I and II) are expressed by fibroblasts in benign and malignant breast lesions, and both are strong mitogens for a number of breast cancer epithelial cell lines in vitro. We have analyzed the stromal mRNA expression of IGF-I and IGF-II in matched sets of fibroblast cell lines derived from three locations in the affected breast of eight patients with breast cancer: (a) the breast tumor itself; (b) surrounding normal breast tissue; and (c) overlying breast skin. IGF-I expression was easily detected in all fibroblasts derived from normal breast tissue. In general, lesser amounts of IGF-I mRNA were detected in fibroblasts derived from breast tumors or skin. In contrast, IGF-II expression was detected at very low levels in only 3 of 8 normal breast fibroblasts, but was present in 6 of 8 tumor fibroblasts. IGF-II mRNA was expressed in all skin fibroblasts tested. IGF-II-negative stromal fibroblasts from normal breast, which were plated at low density and allowed to grow to confluence in the presence of MCF-7 breast tumor epithelial cells, demonstrated a marked increase in IGF-II mRNA expression. IGF-II in situ hybridization studies confirmed that IGF-II expression is seen at high levels in stroma of many invasive breast cancers but not normal breast. We conclude that paracrine influences, mediated by soluble factors released by breast tumor epithelium, are able to specifically increase expression of IGF-II in breast stroma, most likely by a process of clonal selection.


Vulvar Cancer, Version 1.2017, NCCN Clinical Practice Guidelines in Oncology.
Vulvar cancer is a rare gynecologic malignancy. Ninety percent of vulvar cancers are predominantly squamous cell carcinomas (SCCs), which can arise through human papilloma virus (HPV)-dependent and HPV-independent pathways. The NCCN Vulvar Cancer panel is an interdisciplinary group of representatives from NCCN Member Institutions consisting of specialists in gynecological oncology, medical oncology, radiation oncology, and pathology. The NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines) for Vulvar Cancer provide an evidence- and consensus-based approach for the management of patients with vulvar SCC. This manuscript discusses the recommendations outlined in the NCCN Guidelines for diagnosis, staging, treatment, and follow-up.


Sustained activation of autophagy suppresses adipocyte maturation via a lipolysis-dependent mechanism.
Dysregulation of macroautophagy/autophagy is implicated in obesity and insulin resistance. However, it remains poorly defined how autophagy regulates adipocyte development. Using adipose-specific rptor/raptor knockout (KO), atg7 KO and atg7 rptor double-KO mice, we show that inhibiting MTORC1 by RPTOR deficiency led to autophagic sequestration of lipid droplets, formation of LD-containing lysosomes, and elevation of basal and isoproterenol-induced lipolysis in vivo and in primary adipocytes. Despite normal differentiation at an early phase, progressive degradation and shrinkage of cellular LDs and downregulation of adipogenic markers PPARG and PLIN1 occurred in terminal differentiation of rptor KO adipocytes, which was rescued by inhibiting lipolysis or lysosome. In contrast, inactivating autophagy by depletion of ATG7 protected adipocytes against RPTOR deficiency-induced formation of LD-containing lysosomes, LD degradation, and downregulation of adipogenic markers in vitro. Ultimately, atg7 rptor double-KO mice displayed decreased lipolysis, restored adipose tissue development, and upregulated thermogenic gene expression in brown and inguinal adipose tissue compared to RPTOR-deficient mice in vivo. Collectively, our study demonstrates that autophagy plays an important role in regulating adipocyte maturation via a lipophagy and lipolysis-dependent mechanism. ATG7: autophagy related 7; BAT: brown adipose tissue; CEBPB/C/EBPβ: CCAAT enhancer binding protein beta; DGAT1: diacylglycerol O-acyltransferase 1; eWAT: epididymal white adipose tissue; iWAT: inguinal white adipose tissue; KO: knockout; LD: lipid droplet; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; MTORC1: mechanistic target of rapamycin kinase complex 1; PLIN1: perepilin 1; PNPLA2/ATGL: patatin-like phospholipase domain containing 2; PPARG/PPARγ: peroxisome proliferator activated receptor gamma; RPTOR: regulatory associated protein of MTOR complex1; TG: triglyceride; ULK1: unc-51 like kinase 1; UCP1: uncoupling protein 1; WAT: white adipose tissue.


Influenza A virus hemagglutinin trimerization completes monomer folding and antigenicity.
Influenza A virus (IAV) remains an important human pathogen largely because of antigenic drift, the rapid emergence of antibody escape mutants that precludes durable vaccination. The most potent neutralizing antibodies interact with cognate epitopes in the globular "head" domain of hemagglutinin (HA), a homotrimeric glycoprotein. The H1 HA possesses five distinct regions defined by a large number of mouse monoclonal antibodies (MAbs), i.e., Ca1, Ca2, Cb, Sa, and Sb. Ca1-Ca2 sites require HA trimerization to attain full antigenicity, consistent with their locations on opposite sides of the trimer interface. Here, we show that full antigenicity of Cb and Sa sites also requires HA trimerization, as revealed by immunofluorescence microscopy of IAV-infected cells and biochemically by pulse-chase radiolabeling experiments. Surprisingly, epitope antigenicity acquired by HA trimerization persists following acid triggering of the globular domains dissociation and even after proteolytic release of monomeric heads from acid-treated HA. Thus, the requirement for HA trimerization by trimer-specific MAbs mapping to the Ca, Cb, and Sa sites is not dependent upon the bridging of adjacent monomers in the native HA trimer. Rather, complete antigenicity of HA (and, by inference, immunogenicity) requires a final folding step that accompanies its trimerization. Once this conformational change occurs, HA trimers themselves would not necessarily be required to induce a highly diverse neutralizing response to epitopes in the globular domain.


Robust pedestrian detection by combining visible and thermal infrared cameras.
With the development of intelligent surveillance systems, the need for accurate detection of pedestrians by cameras has increased. However, most of the previous studies use a single camera system, either a visible light or thermal camera, and their performances are affected by various factors such as shadow, illumination change, occlusion, and higher background temperatures. To overcome these problems, we propose a new method of detecting pedestrians using a dual camera system that combines visible light and thermal cameras, which are robust in various outdoor environments such as mornings, afternoons, night and rainy days. Our research is novel, compared to previous works, in the following four ways: First, we implement the dual camera system where the axes of visible light and thermal cameras are parallel in the horizontal direction. We obtain a geometric transform matrix that represents the relationship between these two camera axes. Second, two background images for visible light and thermal cameras are adaptively updated based on the pixel difference between an input thermal and pre-stored thermal background images. Third, by background subtraction of thermal image considering the temperature characteristics of background and size filtering with morphological operation, the candidates from whole image (CWI) in the thermal image is obtained. The positions of CWI (obtained by background subtraction and the procedures of shadow removal, morphological operation, size filtering, and filtering of the ratio of height to width) in the visible light image are projected on those in the thermal image by using the geometric transform matrix, and the searching regions for pedestrians are defined in the thermal image. Fourth, within these searching regions, the candidates from the searching image region (CSI) of pedestrians in the thermal image are detected. The final areas of pedestrians are located by combining the detected positions of the CWI and CSI of the thermal image based on OR operation. Experimental results showed that the average precision and recall of detecting pedestrians are 98.13% and 88.98%, respectively.


Treatment with CNX-011-67, a novel GPR40 agonist, delays onset and progression of diabetes and improves beta cell preservation and function in male ZDF rats.
The role of G protein-coupled receptor (GPR40), which is highly expressed in pancreatic beta cells, has been studied extensively in the amelioration of beta cell dysfunction in T2D using rat and mouse islets, beta cell lines and in animal models of diabetes. But its potential as a therapeutic target has not been fully explored. This aim of the study is to evaluate the therapeutic potential of CNX-011-67, a highly selective, potent and orally bioavailable GPR40 agonist, in controlling diabetes and other metabolic parameters. Seven week old male ZDF rats were treated with either vehicle or CNX-011-67, 5 mg/kg twice daily, for seven weeks. The animals were subjected to oral glucose tolerance and insulin tolerance tests. Plasma glucose, insulin, triglyceride, HbA1c, fructosamine and free fatty acids were measured at selected time points. Pancreas from control and treated animals were subjected to insulin and pancreatic and duodenal homeobox 1 (PDX1) immunohistochemistry and were also evaluated by electron microscopy. Also the potential impact of CNX-011-67 on islet insulin secretion, content, ATP levels and markers of both glucose oxidation, beta cell health in rat islets under chronic glucolipotoxic conditions was evaluated. Treatment of male ZDF rats with CNX-011-67 for 7 weeks significantly enhanced insulin secretion in response to oral glucose load, delayed the onset of fasting hyperglycemia by 3 weeks, reduced nonfasting glucose excursions, fasting free fatty acids and triglyceride levels. A significant increase in PDX1 expression and insulin content and reduction in plasma fructosamine, HOMA-IR, and beta cell apoptosis were observed. CNX-011-67 improves glucose mediated insulin secretion, insulin gene transcription and islet insulin content in cultured rat islets under chronic glucolipotoxic condition. Also enhanced glucose oxidation in the form of increased islet ATP content and overall improvement in beta cell health in the form of reduced expression of stress markers (TXNIP and CHOP mRNA) were observed. These findings, suggest that long-term oral therapy with CNX-011-67 could be of clinical value to provide good glycemic control and improve islet beta cell function.


Subchronic and reproductive toxicity of whole dried Hoodia parviflora aerial parts in the rat.
Hoodia parviflora is being developed commercially for use in weight loss food and dietary supplement products. As part of the safety assessment process for H. parviflora, a freeze dried powder preparation was tested in a 90-day oral toxicity study with reproductive/recovery component in rats. Groups of 10 male and female Sprague-Dawley rats were administered H. parviflora dried powder at doses of 0, 100, 250, and 350 mg/kg body weight/day by gavage for an 11-week pre-mating period and a 14-day co-habitation period, and for females, through lactation day 4. An additional 5 rats/sex/group received 0 or 350 mg/kg bw/day for 90 days and were sacrificed 28 days after cessation of treatment. Statistically significant, non-adverse reductions in body weight, body weight gain, food consumption and food efficiency were observed at 250 and 350 mg/kg/day in females. Food consumption was reduced in high-dose males. There were no adverse effects on hematological, blood biochemical, coagulation or urinalysis parameters or on the results of the functional observational battery and histopathological examinations. No evidence of any effect was noted on reproductive or developmental parameters. The NOAEL for dried H. parviflora powder was 350 mg/kg bw/day, the highest permissible dose tested, for both male and female rats.


Parenting styles and emerging adult drug use in Cebu, the Philippines.
Parenting style is a potent and malleable influence on emerging adult substance use. Most of the parenting-substance use literature has been conducted in Western populations and it is unknown whether findings are generalizable to other cultures and contexts. We extended the parenting-substance use literature to a cohort of emerging adults in the Philippines using the Cebu Longitudinal Health and Nutrition Survey. We assessed associations between mothers' and fathers' parenting styles (authoritative, permissive, authoritarian, and neglectful) reported by offspring at age 18 and odds of offspring-reported drug use three years later, adjusted for a range of offspring- and parent/household-level characteristics. Females were dropped from analyses due to low prevalence of drug users. We found that many emerging adults in Cebu reported having used drugs, particularly methamphetamine-a dangerous drug with high abuse potential. Authoritative (warm, firm) mothering was significantly associated with sons' reduced odds of drug use and neglectful fathering was related at a trend level with sons' increased odds of having tried drugs. Findings underscore the relation of parenting styles to emerging adults' drug use and add to the literature on cross-cultural variability in parenting styles.


Lactobacillus jixianensis sp. nov., Lactobacillus baoqingensis sp. nov., Lactobacillus jiayinensis sp. nov., Lactobacillus zhaoyuanensis sp. nov., Lactobacillus lindianensis sp. nov., Lactobacillus huananensis sp. nov., Lactobacillus tangyuanensis sp. nov., Lactobacillus fuyuanensis sp. nov., Lactobacillus tongjiangensis sp. nov., Lactobacillus fujinensis sp. nov. and Lactobacillus mulengensis sp. nov., isolated from Chinese traditional pickle.
Thirty Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle. The strains were characterized using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), determination of average amino acid identity (AAI) and an analysis of phenotypic features. The data demonstrated that the 30 strains represented 11 novel species belonging to the genus Lactobacillus, strains 159-4T, 47-3T, 257-1T, 187-3T, 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T and 112-3T were designated as the type strains. Strains 159-4T and 47-3T were related to the type strains of Lactobacillus porcinae, Lactobacillus manihotivorans, Lactobacillus nasuensis, Lactobacillus camelliae, Lactobacillus pantheris, Lactobacillus thailandensis, Lactobacillus sharpeae and Lactobacillus songhuajiangensis, having 92.0-98.9 % 16S rRNA gene sequence similarities, 58.1-84.7 % pheS gene sequence similarities and 61.4-90.3 % rpoA gene sequence similarities. Strains 257-1T and 187-3T were related to the type strains of Lactobacillus coryniformis, Lactobacillus iwatensis, Lactobacillus backii, Lactobacillus rennini and Lactobacillus bifermentans, having 93.5-99.3 % 16S rRNA gene sequence similarities, 67.7-81.8 % pheS gene sequence similarities and 77.0-96.2 % rpoA gene sequence similarities. Strains 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T and 112-3T were closely related to the type strains of Lactobacillus paucivorans, Lactobacillus brevis, Lactobacillus hammesii, Lactobacillus senmaizukei, Lactobacillus parabrevis, Lactobacillus yonginensis, Lactobacillus koreensis and Lactobacillus cerevisiae, having 95.6-99.9 % 16S rRNA gene sequence similarities, less than 93.9 % pheS gene sequence similarities and 87.0-99.7 % rpoA gene sequence similarities. ANI, isDDH and AAI values between strains 159-4T, 47-3T, 257-1T, 187-3T, 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T, 112-3T and type strains of phylogenetically related species were less than 92.7, 48.4 and 96.6 %, respectively, confirming that they represent 11 novel species within the genus Lactobacillus. Based upon the data of polyphasic characterization obtained in the present study, eleven novel species, Lactobacillusjixianensis sp. nov., Lactobacillusbaoqingensis sp. nov., Lactobacillusjiayinensis sp. nov., Lactobacilluszhaoyuanensis sp. nov., Lactobacilluslindianensis sp. nov., Lactobacillushuananensis sp. nov., Lactobacillustangyuanensis sp. nov., Lactobacillusfuyuanensis sp. nov., Lactobacillustongjiangensis sp. nov., Lactobacillusfujinensis sp. nov. and Lactobacillusmulengensis sp. nov., are proposed and the type strains are 159-4T (=NCIMB 15175T=CCM 8911T), 47-3T (=NCIMB 15165T=CCM 8903T=LMG 31064T), 257-1T (=NCIMB 15166T=CCM 8904T=LMG 31065T), 187-3T (=NCIMB 15172T =CCM 8910T), 220-4T (=NCIMB 15163T =CCM 8902T=KCTC 21136T), 151-2BT (=NCIMB 15164T=CCM 8913T=KCTC 21129T=LMG 31063T), 137-3T (=NCIMB 15170T=CCM 8907T=KCTC 21125T=LMG 31053T), 244-4T (=NCIMB 15168T=CCM 8906T=KCTC 21137T=LMG 31052T), 218-10T (=NCIMB 15167T=CCM 8905T=KCTC 21135T =LMG 31055T), 218-6T (=NCIMB 15171T=CCM 8908T=KCTC 21134T =LMG 31067T) and 112-3T (=NCIMB 15174T=CCM 8909T=KCTC 21123T=LMG 31049T), respectively.


Atomic scale microstructure and properties of Se-deficient two-dimensional MoSe2.
We study the atomic scale microstructure of nonstoichiometric two-dimensional (2D) transition metal dichalcogenide MoSe2-x by employing aberration-corrected high-resolution transmission electron microscopy. We show that a Se-deficit in single layers of MoSe2 grown by molecular beam epitaxy gives rise to a dense network of mirror-twin-boundaries (MTBs) decorating the 2D-grains. With the use of density functional theory calculations, we further demonstrate that MTBs are thermodynamically stable structures in Se-deficient sheets. These line defects host spatially localized states with energies close to the valence band minimum, thus giving rise to enhanced conductance along straight MTBs. However, electronic transport calculations show that the transmission of hole charge carriers across MTBs is strongly suppressed due to band bending effects. We further observe formation of MTBs during in situ removal of Se atoms by the electron beam of the microscope, thus confirming that MTBs appear due to Se-deficit, and not coalescence of individual grains during growth. At a very high local Se-deficit, the 2D sheet becomes unstable and transforms to a nanowire. Our results on Se-deficient MoSe2 suggest routes toward engineering the properties of 2D transition metal dichalcogenides by deviations from the stoichiometric composition.


Interfacial elastic fingering in Hele-Shaw cells: a weakly nonlinear study.
We study a variant of the classic viscous fingering instability in Hele-Shaw cells where the interface separating the fluids is elastic, and presents a curvature-dependent bending rigidity. By employing a second-order mode-coupling approach we investigate how the elastic nature of the interface influences the morphology of emerging interfacial patterns. This is done by focusing our attention on a conventionally stable situation in which the fluids involved have the same viscosity. In this framework, we show that the inclusion of nonlinear effects plays a crucial role in inducing sizable interfacial instabilities, as well as in determining the ultimate shape of the pattern-forming structures. Particularly, we have found that the emergence of either narrow or wide fingers can be regulated by tuning a rigidity fraction parameter. Our weakly nonlinear findings reinforce the importance of the so-called curvature weakening effect, which favors the development of fingers in regions of lower rigidity.


Evidence for regulation of monomethyl mercury by nitrate in a seasonally stratified, eutrophic lake.
The accumulation of monomethyl mercury (CH3Hg+) in aquatic ecosystems is a redox sensitive process that is accelerated under sulfate-reducing conditions. While nitrate (NO3-) reduction is energetically favored over sulfate reduction, the influence of NO3 on the accumulation of CH3Hg+ has not been reported in the literature. We examined temporal and vertical patterns in redox constituents and CH3Hg+ concentrations in the hypolimnion of a dimictic lake, Onondaga Lake, prior to and following increases in NO3- inputs. Detailed water-column profiles and a long-term record revealed marked decreases in the accumulation of CH3Hg+ in the anoxic hypolimnion coinciding with long-term decreases in the deposition of organic matter coupled with recent increases in NO3-concentrations. CH3Hg+ concentrations in the hypolimnion were substantially abated when No3 was present above the sediment-water interface. A decrease in the peak hypolimnetic mass of CH3Hg+ and shortening of the period of elevated CH3Hg+ concentrations resulted in more than a 50% decline in the accumulated CH3Hg+. N03- regulation of CH3Hg+ accumulation may be a widespread phenomenon in oxygen-limited freshwater and terrestrial environments, and could have an important notpreviously recognized, effect on the biogeochemistry of mercury.


Cytological evaluation of the nasal mucosa in neonates exposed to tobacco smoke during fetal life.
The objective of this study is to assess the cytological picture of the nasal mucosa of neonates born to mothers who are active smokers, passive smokers and non-smokers. A prospective study was conducted in a group of 86 neonates born between 23 and 41 weeks of gestation. The assignation of neonates to one of the three aforementioned groups was based on a questionnaire concerning exposure to tobacco smoke, and on the concentration of cotinine in maternal urine. A cytological examination was performed using exfoliative cytology with a semi-quantitative evaluation of the cells present in the specimens. Hematological summation equipment was used to assess the number of neutrophils, eosinophils, columnar, goblet, basal and squamous cells out of 500 cells counted. The number of specific cells was expressed as a percentage and a cytogram was created. The most common type of cytogram contained neutrophils, columnar cells, and squamous cells. No significant differences were observed between the subgroups. Similarly, there was no correlation between the median of each type of cell and the cotinine concentration in the mothers' urine. Active and passive smoking during pregnancy do not influence the cytological picture of the nasal mucosa of neonates.


Suppurative iliac lymphadenitis masquerading as appendicitis in an HIV-infected patient.
An HIV-infected man presented with acute onset of right iliac fossa pain. Initial assessment of clinical and computed tomography findings suggested a diagnosis of appendicitis. A macroscopically normal appendix was removed. Further imaging performed postoperatively because of persistent symptoms showed a right iliac fossa mass; culture of pus aspirated from the mass grew group A beta-haemolytic Streptococcus confirming a diagnosis of suppurative iliac lymphadenitis.


A Neanderthal Sodium Channel Increases Pain Sensitivity in Present-Day Humans.
The sodium channel Nav1.7 is crucial for impulse generation and conduction in peripheral pain pathways [1]. In Neanderthals, the Nav1.7 protein carried three amino acid substitutions (M932L, V991L, and D1908G) relative to modern humans. We expressed Nav1.7 proteins carrying all combinations of these substitutions and studied their electrophysiological effects. Whereas the single amino acid substitutions do not affect the function of the ion channel, the full Neanderthal variant carrying all three substitutions, as well as the combination of V991L with D1908G, shows reduced inactivation, suggesting that peripheral nerves were more sensitive to painful stimuli in Neanderthals than in modern humans. We show that, due to gene flow from Neanderthals, the three Neanderthal substitutions are found in ∼0.4% of present-day Britons, where they are associated with heightened pain sensitivity.


Impact of maternal anxiety level on the childhood vaccination coverage.
The mother's mental state as a risk factor for the children's vaccination status in developing countries has received little attention. The aim of this study was to determine the association between childhood vaccination coverage and maternal anxiety. A total of 195 consecutive infants and their mothers attending a tertiary teaching hospital, department of pediatric outpatient center between January 2008 and September 2009 were included in the study. One hundred five infants who have incomplete vaccination schedule (according to the National Immunization Schedule) were matched with 90 controls (fully vaccinated) and their mothers self-report measure of anxiety level using the State-Trait Anxiety Inventory (STAI), a psychiatric screening instrument. The chi-square test and the logistic regression were used in the statistical analysis. High maternal anxiety levels determined by STAI was associated with increased risk of incomplete vaccination status in infants (odds ratio 4.35, 95% confidence interval 1.87-8.79).This association remained significant after controlling for sociodemographic factors. High maternal anxiety scores may result in incomplete vaccination status in children younger than 3 years.


Well-being therapy. A novel psychotherapeutic approach for residual symptoms of affective disorders.
There is increasing awareness of the prognostic value of residual symptomatology in affective disorders and of the need for specific therapeutic strategies in this phase of illness. The aims of the study were to apply a novel, short-term psychotherapeutic approach for increasing well-being, based on Ryff's conceptual model, to remitted patients with affective disorders and to compare the results with those obtained with symptom-oriented cognitive behavioural strategies. Twenty patients with affective disorders (major depression, panic disorder with agoraphobia, social phobia, generalized anxiety disorder, obsessive-compulsive disorder) who had been successfully treated by behavioural or pharmacological methods were randomly assigned to a well-being enhancing therapeutic strategy (well-being therapy) or cognitive-behavioural treatment of residual symptoms. Both well-being and cognitive-behavioural therapies were associated with a significant reduction of residual symptoms. However, a significant advantage of well-being therapy over cognitive-behavioural strategies was observed with observer-rated methods. These preliminary results suggest the feasibility of well-being therapy in the residual stage of affective disorders. Further research should determine its value as a relapse-preventive strategy in specific mood and anxiety disorders.


Effect of prenatal administration of venlafaxine on postnatal development of rat offspring.
About 3% of pregnant women are treated with antidepressant drugs during gestation. After delivery the number of treated women increases to 5 to 7%. Most prescribed antidepressants in pregnancy are selective serotonin re-uptake inhibitors and/or serotonin and noradrenaline re-uptake inhibitors, such as fluoxetine, paroxetine, sertraline, citalopram and venlafaxine (VENF). Despite the fact that VENF has been assigned to pregnancy category C by the FDA, experimental studies with this drug are rare. The aim of this pilot study was to investigate the effect of prenatal administration of VENF on early postnatal development of rat offspring and selected biochemical variables at weaning of pups. Pregnant female Wistar rats were treated with VENF from day 15 to 20 of gestation at the doses of 7.5, 37.5 and 70 mg/kg. Females were allowed to spontaneously deliver their pups. After delivery the pups were inspected for viability, gross malformation and they were weighed on day 0, 4 and 21 post partum. On day 21 post partum, the pups were killed, brains were removed from the skulls and blood samples were collected for biochemical assay (proteins, glucose-GOD, glucose-HEX, lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase and total antioxidant status). The study showed that prenatal VENF administration resulted in a mild maternal intoxication manifested by decreased body weight gain of pregnant females. There was no effect of the drug tested on the body and brain weights of offspring. No obvious morphological alterations were observed in the delivered pups. Similarly, there were no changes in the selected biochemical variables determined.


Aortic Coarctation Treated by PTA and Stenting: a Case Presentation and Literature Review.
A 13 year-old boy was diagnosed with simple aortic coarctation and a bicuspid aortic valve. Ascending aorta was mildly dilated while aortic arch was smaller, but not hypoplastic. A decision to treat him was made depending on the clinical finding of systemic hypertension in his young age and a baseline trans-stenotic echographic gradient of 48 mmHg. Direct stenting with a covered Cheetham-Platinum balloon-expandable stent was performed with no complications. Despite immediate good angiographic and hemodynamic result with marked decrease of invasively measured gradient, consecutive echographic follow-up demonstrated progressive increase of the gradient in the first 9 months after the index procedure. This resulted in repeated aortic PTA with a larger balloon inside the previously implanted stent-graft. Optimal angiographic result was obtained, the patient remained symptom-free and no antihypertensive therapy was necessary thereafter. Modern treatment of simple aortic coarctation can be done in the cath-lab in appropriately selected cases. Stent implantation should be considered in all cases to avoid dissection and early elastic recoil. Long term follow-up may evidentiate the need for repeated PTA procedures in some cases.


Transcriptome and translational signaling following endurance exercise in trained skeletal muscle: impact of dietary protein.
Postexercise protein feeding regulates the skeletal muscle adaptive response to endurance exercise, but the transcriptome guiding these adaptations in well-trained human skeletal muscle is uncharacterized. In a crossover design, eight cyclists ingested beverages containing protein, carbohydrate and fat (PTN: 0.4, 1.2, 0.2 g/kg, respectively) or isocaloric carbohydrate and fat (CON: 1.6, 0.2 g/kg) at 0 and 1 h following 100 min of cycling. Biopsies of the vastus lateralis were collected at 3 and 48 h following to determine the early and late transcriptome and regulatory signaling responses via microarray and immunoblot. The top gene ontology enriched by PTN were: muscle contraction, extracellular matrix--signaling and structure, and nucleoside, nucleotide, and nucleic acid metabolism (3 and 48 h); developmental processes, immunity, and defense (3 h); glycolysis, lipid and fatty acid metabolism (48 h). The transcriptome was also enriched within axonal guidance, actin cytoskeletal, Ca2+, cAMP, MAPK, and PPAR canonical pathways linking protein nutrition to exercise-stimulated signaling regulating extracellular matrix, slow-myofibril, and metabolic gene expression. At 3 h, PTN attenuated AMPKα1Thr172 phosphorylation but increased mTORC1Ser2448, rps6Ser240/244, and 4E-BP1-γ phosphorylation, suggesting increased translation initiation, while at 48 h AMPKα1Thr172 phosphorylation and PPARG and PPARGC1A expression increased, supporting the late metabolic transcriptome, relative to CON. To conclude, protein feeding following endurance exercise affects signaling associated with cell energy status and translation initiation and the transcriptome involved in skeletal muscle development, slow-myofibril remodeling, immunity and defense, and energy metabolism. Further research should determine the time course and posttranscriptional regulation of this transcriptome and the phenotype responding to chronic postexercise protein feeding.


Vancomycin prescription in neonates and young infants: toward a simplified dosage.
There is no consensus on vancomycin dosing in newborns and young infants. The first objective was to assess the efficiency of a simplified dosing regimen with a cohort study. The secondary objective was to examine pharmacokinetic data to determine how this simplified dosing could be improved. All neonates admitted to our intensive care unit and treated with vancomycin were included in the pharmacokinetic study (PK group, 83 treatments, 156 measurements). The vancomycin dosing regimen consisted of a loading dose of 7 mg/kg, followed by a constant continuous dose of 30 mg/kg/day. The target serum vancomycin concentration ranged from 10 mg/l to 30 mg/l. Data from patients whose medications followed the scheduled dosing without modifications or prescription errors (actual dosing group: 62 treatments, 108 measurements) were analysed separately. A population pharmacokinetic analysis was performed (PK group) to simulate several vancomycin dosings. Prescription errors were found in 10 of 83 treatments (12%). In the actual dosing group, 89.2% of vancomycin measurements were within the target range. Serum creatinine remained stable throughout treatment. Vancomycin concentrations varied widely. The modified regimen for a target vancomycin concentration of 25 mg/l consisted of a bolus of 20 mg/kg followed by continuous infusion of 30 mg/kg. Our pharmacokinetic data and bedside results suggest that a simplified schedule of vancomycin can achieve the targeted drug concentrations in most patients while avoiding secondary renal toxicity. The proposed new dosing scheme should be validated in a drug survey, but due to pharmacokinetic variability, still requires therapeutic drug monitoring.


Estimating the Risk of Pneumonia in Patients With Schizophrenia Newly Receiving Clozapine: A Nationwide Cohort Study.
Patients with schizophrenia are vulnerable to pneumonia. Clozapine is associated with the greatest risk of pneumonia. We investigated the risk factors of pneumonia in patients with schizophrenia who use clozapine. We used a large cohort of patients with schizophrenia (N = 22,774) who newly use clozapine (baseline). We divided the data set into a training cohort (entry between 1998 and 2008, n = 18,496) and test cohort (entry between 2009 and 2012, n = 4278), where 483 and 168 patients developed pneumonia requiring hospitalization within 1 year after baseline, respectively. For prediction, we developed a static model using Cox proportional hazards regression and a dynamic model using Cox regression with time-dependent modeling. Areas under receiver operating curves (AUCs) for the predictive model were estimated in the training cohort and then in the test cohort for validation. Based on the baseline characteristics, the static model for predicting pneumonia in 3 periods (90, 180, and 365 days) was unsatisfactory (AUCs, 0.64, 0.64, and 0.65, respectively). The predictors were older age, male sex, history of nonpsychiatric hospitalization, dementia, asthma, and tuberculosis within 1 year before baseline. However, the results were improved (AUCs, 0.83, 0.79, and 0.77, respectively) after control for time-dependent variables, namely, duration of clozapine use and concomitant medications (ie, benzodiazepines, valproic acid, systemic corticosteroids). Several risk factors for predicting subsequent pneumonia after initial use of clozapine were explored, including older age, male, history of nonpsychiatric hospitalization, dementia, asthma, tuberculosis, benzodiazepines, valproic acid, systemic corticosteroids, and the use duration of clozapine. Clinical staff can use the risk factors to administer evidence-based treatment.


Brain systems for probabilistic and dynamic prediction: computational specificity and integration.
A computational approach to functional specialization suggests that brain systems can be characterized in terms of the types of computations they perform, rather than their sensory or behavioral domains. We contrasted the neural systems associated with two computationally distinct forms of predictive model: a reinforcement-learning model of the environment obtained through experience with discrete events, and continuous dynamic forward modeling. By manipulating the precision with which each type of prediction could be used, we caused participants to shift computational strategies within a single spatial prediction task. Hence (using fMRI) we showed that activity in two brain systems (typically associated with reward learning and motor control) could be dissociated in terms of the forms of computations that were performed there, even when both systems were used to make parallel predictions of the same event. A region in parietal cortex, which was sensitive to the divergence between the predictions of the models and anatomically connected to both computational networks, is proposed to mediate integration of the two predictive modes to produce a single behavioral output.


Aberrant RNA splicing and mutations in spliceosome complex in acute myeloid leukemia.
The spliceosome, the cellular splicing machinery, regulates RNA splicing of messenger RNA precursors (pre-mRNAs) into maturation of protein coding RNAs. Recurrent mutations and copy number changes in genes encoding spliceosomal proteins and splicing regulatory factors have tumor promoting or suppressive functions in hematological malignancies, as well as some other cancers. Leukemia stem cell (LSC) populations, although rare, are essential contributors of treatment failure and relapse. Recent researches have provided the compelling evidence that link the erratic spicing activity to the LSC phenotype in acute myeloid leukemia (AML). In this article, we describe the diverse roles of aberrant splicing in hematological malignancies, particularly in AML and their contributions to the characteristics of LSC. We review these promising strategies to exploit the addiction of aberrant spliceosomal machinery for anti-leukemic therapy with aim to eradicate LSC. However, given the complexity and plasticity of spliceosome and not fully known functions of splicing in cancer, the challenges facing the development of the therapeutic strategies targeting RAN splicing are highlighted and future directions are discussed too.


Intratracheally administered 5-azacytidine is effective against orthotopic human lung cancer xenograft models and devoid of important systemic toxicity.
Hypermethylation of key tumor suppressor genes plays an important role in lung carcinogenesis. The purpose of this study is to explore the therapeutic potential of regional administration (via the airways) of the demethylating agent 5-azacytidine (5-Aza) for the treatment of early lung cancer. We administered 5-Aza solution directly into the trachea in imprinting control region (ICR) mice (to study its toxicity) and in nude mice bearing orthotopic human lung cancer xenografts (to assess its antitumor activity). In vitro, 5-Aza inhibited the growth of human lung cancer cell lines H226, H358, and H460 in a dose-dependent manner. The concentrations to inhibit cell growth by 50% (IC50) were about 0.6-4.9 µg/mL. 5-Azacytidine reversed hypermethylation in the promoter of tumor suppressor gene RASSF1a in the H226 cells at a 6000-fold lower concentration than its IC50. In animal studies, intratracheal (I.T.) administration of 90 mg/kg 5-Aza (the maximum tolerated dose of 5-Aza intravenous injection [I.V.]) resulted in moderate pulmonary toxicity and 5-fold reduced myelosuppression compared with the same dose of I.V. 5-Aza. Using an optimized multiple dose schedule, I.T. 5-Aza was about 3-fold more effective than I.V. 5-Aza in prolonging the survival of mice bearing orthotopic H460 and H358 xenografts, and did not cause any detectable toxicity. 5-Azacytidine can reverse the hypermethylation in the human lung cancer cell lines at a nontoxic dose. Regional administration to the airways enhances the therapeutic index of 5-Aza by 75-fold. The potential of regional administration of 5-Aza (including by aerosolization) for the treatment of advanced bronchial premalignancy deserves further investigation.


Structure elucidation and preliminary assessment of hydrolase activity of PqsE, the Pseudomonas quinolone signal (PQS) response protein.
In bacteria, the transcription of virulence genes is usually controlled by a cell density-dependent process known as "quorum sensing" (QS). QS relies on small diffusible signaling molecules that cross the bacterial cell wall and activate target transcription factors after a threshold concentration has been reached. Besides two hierarchical QS circuits based on N-acylhomoserine lactones, the human opportunistic pathogen Pseudomonas aeruginosa integrates a signaling system that depends on 2-heptyl-3-hydroxy-4-quinolone, termed "Pseudomonas quinolone signal" (PQS). PQS is produced from genes encoded in the pqs operon, which in addition to the biosynthetic enzymes PqsA-D contains a fifth gene, pqsE, that is not required for production of PQS but whose disruption leads to loss of signal transduction in several but not all pqs operon-dependent processes. PqsE was hence termed "PQS response protein", but its exact mechanism of action is unknown. We have determined the crystal structure of recombinant PqsE and show that it possesses a metallo-beta-lactamase fold with an Fe(II)Fe(III) center in the active site. A copurified ligand was assigned as benzoate and may indicate that PqsE exerts its regulatory effect by converting a chorismate-derived molecule. Further, PqsE was found to slowly hydrolyze phosphodiesters including single- and double-stranded DNA as well as mRNA and also the thioester S-(4-nitrobenzoyl)mercaptoethane. Higher activity was observed after incubation with Co(2+) and, to lesser entent, Mn(2+), suggesting that the Fe(II)Fe(III) center of recombinant PqsE may be an artifact of heterologous expression. A crystal complex of the E182A mutant with bis-pNPP was obtained and suggests a catalytic mechanism for hydrolysis.


Defining the human adipose tissue proteome to reveal metabolic alterations in obesity.
White adipose tissue (WAT) has a major role in the progression of obesity. Here, we combined data from RNA-Seq and antibody-based immunohistochemistry to describe the normal physiology of human WAT obtained from three female subjects and explored WAT-specific genes by comparing WAT to 26 other major human tissues. Using the protein evidence in WAT, we validated the content of a genome-scale metabolic model for adipocytes. We employed this high-quality model for the analysis of subcutaneous adipose tissue (SAT) gene expression data obtained from subjects included in the Swedish Obese Subjects Sib Pair study to reveal molecular differences between lean and obese individuals. We integrated SAT gene expression and plasma metabolomics data, investigated the contribution of the metabolic differences in the mitochondria of SAT to the occurrence of obesity, and eventually identified cytosolic branched-chain amino acid (BCAA) transaminase 1 as a potential target that can be used for drug development. We observed decreased glutaminolysis and alterations in the BCAAs metabolism in SAT of obese subjects compared to lean subjects. We also provided mechanistic explanations for the changes in the plasma level of BCAAs, glutamate, pyruvate, and α-ketoglutarate in obese subjects. Finally, we validated a subset of our model-based predictions in 20 SAT samples obtained from 10 lean and 10 obese male and female subjects.


The biological clock modulates the human cortisol response in a multiplicative fashion.
Human cortisol levels follow a clear circadian rhythm. We investigated the contribution of alternation of sleep and wakefulness and the circadian clock, using forced desynchrony. Cortisol levels were best described by a multiplication of a circadian and a wake-time component. The human cortisol response is modulated by circadian phase. Exposure to stress at an unnatural phase, as in shift work, is predicted to result in abnormal cortisol levels. Health of shift workers may therefore improve when stress is reduced at times when the clock produces high stress sensitivity.


[The Montreal definition and classification of gastroesophageal reflux disease: a global, evidence-based consensus paper].
A world-wide recognised and accepted definition and classification of gastroesophageal reflux disease (GERD) would be highly desirable for research and clinical practice. The purpose of this project was to develop such a generally accepted definition and classification that could be used equally by patients, physicians, and supervisory bodies. In order to ensure a consensus among the participating experts a modified delphi process with a step-wise selection modality was employed. For this the working group of five persons formulated a series of statements on the basis of a systematic search of the literature using three databases (Embase, Cochrane-Study register, Medline). Then these statements were developed further for two years, revised and finally passed as consensus. The consensus group consisted of 44 experts from 18 countries. Each key vote was held on the basis of a six-point scale. A "consensus" was considered to have been reached when two-thirds of the participants voted in favour of the respective statement. The level of agreement between the experts increased in the course of the multistep decision process, in the individual voting steps requiring at least two-thirds of the participants, the results were at first 86%, then 88% through to 94% and finally 100% in favour of the chosen statement. In the final voting, 94% of the final 51 statements were accepted by 90% of the consensus group. 90% of all statements were accepted unanimously or with only minor reservations. GERD was defined as a disease that is associated with troublesome symptoms and/or complications on account of reflux of stomach contents into the esophagus. The complaints are divided into esophageal and extra-esophageal syndromes. Among the novel aspects of this definition are the patient-orientated approach that is independent of endoscopic findings, the classification of the ailment into independent syndromes as well as the consideration of laryngitis, cough, asthma and dental problems as possible GERD syndromes. Furthermore, a new definition of suspected or demonstrated Barrett's esophagus is proposed. Irrespective of country-specific differences in terminology, language, prevalence and manifestations of this disease, evidence-based, world-wide valid consensus definitions are possible. A global consensus definition of GERD will simplify disease management, make mutual research possible and help in the design of generally valid studies. This will not only help the patient but also the physician and supervisory bodies.


Identification of a candidate regulatory region in the human CD8 gene complex by colocalization of DNase I hypersensitive sites and matrix attachment regions which bind SATB1 and GATA-3.
To locate elements regulating the human CD8 gene complex, we mapped nuclear matrix attachment regions (MARs) and DNase I hypersensitive (HS) sites over a 100-kb region that included the CD8B gene, the intergenic region, and the CD8A gene. MARs facilitate long-range chromatin remodeling required for enhancer activity and have been found closely linked to several lymphoid enhancers. Within the human CD8 gene complex, we identified six DNase HS clusters, four strong MARs, and several weaker MARs. Three of the strong MARs were closely linked to two tissue-specific DNase HS clusters (III and IV) at the 3' end of the CD8B gene. To further establish the importance of this region, we obtained 19 kb of sequence and screened for potential binding sites for the MAR-binding protein, SATB1, and for GATA-3, both of which are critical for T cell development. By gel shift analysis we identified two strong SATB1 binding sites, located 4.5 kb apart, in strong MARs. We also detected strong GATA-3 binding to an oligonucleotide containing two GATA-3 motifs located at an HS site in cluster IV. This clustering of DNase HS sites and MARs capable of binding SATB1 and GATA-3 at the 3' end of the CD8B gene suggests that this region is an epigenetic regulator of CD8 expression.


Clinical appropriateness: a key factor in outcome measure selection: the 36 item short form health survey in multiple sclerosis.
Understanding the properties of an outcome measure is essential in choosing the appropriate instrument and interpreting the information it generates. The MOS 36 item short form health survey questionnaire (SF-36) is widely acknowledged as the gold standard generic measure of health status; few studies however have evaluated its use for clinical trials in multiple sclerosis. Its clinical appropriateness, internal consistency reliability, validity, and responsiveness was investigated across a broad range of patients with multiple sclerosis. A prospective study in which 150 adults with clinically definite multiple sclerosis completed a battery of questionnaires evaluating generic health status, disability, handicap, and emotional wellbeing. Of these, 44 patients undergoing inpatient rehabilitation completed the questionnaires before and after intervention to evaluate responsiveness. Score distributions demonstrated significant floor and ceiling effects in four of the eight dimensions which were particularly marked when patient selection was restricted to a narrow band of disease severity (as is the case in most clinical trials). Internal consistency exceeded the standard for group comparisons for all dimensions. Convergent and discriminant construct validity was supported by the direction, magnitude, and pattern of correlations with other health measures. In comparison with instruments measuring associated constructs, the responsiveness of the SF-36 was poor in evaluating change in moderate to severely disabled patients participating in a programme of inpatient rehabilitation. The SF-36 has some limitations as an outcome measure in multiple sclerosis. The results highlight the need for all instruments to be examined in the specific sample population under question and for the specific research question being investigated. In multiple sclerosis clinical trials, the SF-36 should be supplemented with other relevant measures.


Bromelain activates murine macrophages and natural killer cells in vitro.
The innate immune response is critical for effective immunity against most pathogens. In this study, we show that bromelain, a mixture of cysteine proteases, can enhance IFN-gamma-mediated nitric oxide and TNFalpha production by macrophages. Bromelain's effect was independent of endotoxin receptor activation and was not caused by direct modulation of IFN-gamma receptors. Instead, bromelain either enhanced or acted synergistically with IFN-gamma receptor-mediated signals. These effects were seen in both RAW 264.7, a macrophage cell line, and primary macrophage populations. Bromelain also increased IL-2- and IL-12-mediated IFN-gamma production by NK cells. These results indicate a potential role for bromelain in the activation of inflammatory responses in situations where they may be deficient, such as may occur in immunocompromised individuals.


Microarray analysis identifies keratin loci as sensitive biomarkers for thyroid hormone disruption in the salamander Ambystoma mexicanum.
Ambystomatid salamanders offer several advantages for endocrine disruption research, including genomic and bioinformatics resources, an accessible laboratory model (Ambystoma mexicanum), and natural lineages that are broadly distributed among North American habitats. We used microarray analysis to measure the relative abundance of transcripts isolated from A. mexicanum epidermis (skin) after exogenous application of thyroid hormone (TH). Only one gene had a >2-fold change in transcript abundance after 2 days of TH treatment. However, hundreds of genes showed significantly different transcript levels at days 12 and 28 in comparison to day 0. A list of 123 TH-responsive genes was identified using statistical, BLAST, and fold level criteria. Cluster analysis identified two groups of genes with similar transcription patterns: up-regulated versus down-regulated. Most notably, several keratins exhibited dramatic (1000 fold) increases or decreases in transcript abundance. Keratin gene expression changes coincided with morphological remodeling of epithelial tissues. This suggests that keratin loci can be developed as sensitive biomarkers to assay temporal disruptions of larval-to-adult gene expression programs. Our study has identified the first collection of loci that are regulated during TH-induced metamorphosis in a salamander, thus setting the stage for future investigations of TH disruption in the Mexican axolotl and other salamanders of the genus Ambystoma.


Nematicons in planar cells subject to the optical Fréedericksz threshold.
We investigate, both theoretically and experimentally, self-trapping of light beams in nematic liquid crystals arranged so as to exhibit the optical Fréedericksz transition in planar cells. The resulting threshold in the nonlinear reorientational response supports a bistable behavior between diffracting and self-localized beam states, leading to the appearance of a hysteretic loop versus input excitation. Our results confirm the role of nematic liquid crystals in the study of non-perturbative nonlinear photonics.


The effect of a tooth/tongue gel and mouthwash regimen on morning oral malodour: A 3-week single-blind randomized clinical trial.
To compare the effects of a regimen consisting of a tooth/tongue gel, tongue cleaner and mouthwash with the effects of using standard fluoride dentifrice on the organoleptic oral malodour score (ORG) and volatile sulphur compounds (VSCs). A total, 66 non-dental students participated in a 3-week parallel, single-blind, randomized, controlled clinical trial. The test group used a tongue cleaner, a tooth/tongue gel and mouthwash containing amine fluoride/stannous fluoride and zinc lactate as oral malodour counteractive. The control group used a standard fluoride dentifrice. Measurements were taken in the morning at baseline, at days 1, 7 and 21. The primary outcome was the ORG score. The secondary outcome, the VSC measurement, was assessed using OralChroma™ (H2 S, CH3 SH, (CH3 )2 S) and Halimeter® . Tongue coating thickness and tongue discoloration were scored. At baseline and day 21, the participants' self-perceptions were assessed. At day 1 for the ORG, H2 S, CH3 SH and Halimeter® readings, a significant decrease was observed in the test group. At day 21, the decrease in H2 S and the Halimeter® outcomes were maintained for the test group, and a significant increase in tongue surface discoloration was observed. The test group evaluated their "morning breath upon awakening" as significantly better (P=.001) after 21 days. A significant overnight effect on morning oral malodour was observed for most of the parameters in favour of the test group. At day 21, the effect of prolonged use was significant for H2 S and the Halimeter® readings, although not for the primary ORG outcome parameter.


A novel low-molecular weight inhibitor of focal adhesion kinase, TAE226, inhibits glioma growth.
Glioblastomas are highly lethal cancers that resist current therapies. Novel therapies under development target molecular mechanisms that promote glioblastoma growth. In glioblastoma patient specimens, the non-receptor tyrosine kinase focal adhesion kinase (FAK) is overexpressed. Upon growth factor receptor stimulation or integrin engagement, FAK is activated by phosphorylation on critical tyrosine residues. Activated FAK initiates a signal transduction cascade which promotes glioma growth and invasion by increasing cellular adhesion, migration, invasion, and proliferation. We find that human glioma cell lines express different levels of total FAK protein and activating phosphorylation of tyrosine residues Tyr397, Tyr861, and Tyr925. As all glioma cell lines examined expressed phosphorylated FAK, we examined the efficacy of a novel low-molecular weight inhibitor of FAK, TAE226, against human glioma cell lines. TAE226 inhibited the phosphorylation of FAK as well as the downstream effectors AKT, extracellular signal-related kinase, and S6 ribosomal protein in multiple glioma cell lines. TAE226 induced a concentration-dependent decrease in cellular proliferation with an associated G(2) cell cycle arrest in every cell line and an increase in apoptosis in a cell-line-specific manner. TAE226 also decreased glioma cell adhesion, migration, and invasion through an artificial extracellular matrix. Together, these data demonstrate the potential benefit of TAE226 for glioma therapy.


Fabrication and evolution of multilayer silver nanofilms for surface-enhanced Raman scattering sensing of arsenate.
Surface-enhanced Raman scattering (SERS) has recently been investigated extensively for chemical and biomolecular sensing. Multilayer silver (Ag) nanofilms deposited on glass slides by a simple electroless deposition process have been fabricated as active substrates (Ag/GL substrates) for arsenate SERS sensing. The nanostructures and layer characteristics of the multilayer Ag films could be tuned by varying the concentrations of reactants (AgNO3/BuNH2) and reaction time. A Ag nanoparticles (AgNPs) double-layer was formed by directly reducing Ag+ ions on the glass surfaces, while a top layer (3rd-layer) of Ag dendrites was deposited on the double-layer by self-assembling AgNPs or AgNPs aggregates which had already formed in the suspension. The SERS spectra of arsenate showed that characteristic SERS bands of arsenate appear at approximately 780 and 420 cm-1, and the former possesses higher SERS intensity. By comparing the peak heights of the approximately 780 cm-1 band of the SERS spectra, the optimal Ag/GL substrate has been obtained for the most sensitive SERS sensing of arsenate. Using this optimal substrate, the limit of detection (LOD) of arsenate was determined to be approximately 5 μg·l-1.


Structured robot-assisted surgery training curriculum for residents in Urology and impact on future surgical activity.
To gain insight into the availability of training for robot assisted surgery (RAS) and the possibility to perform RAS during Dutch residency curriculum and to analyze the effects on surgical skills by the introduction of an advanced course in RAS for residents. A combination of a validated snap shot survey and a prospective cohort study. Structured advanced RAS training including virtual reality (VR) simulation, dry and wet lab facility at ORSI academy (Belgium). A snap-shot survey has been sent to all the residents and specialists in Urology graduated during the years 2017-2020 in Netherlands. Among residents, only last year residents (5th and 6th year) have been considered for the RAS training. Although most of the residents (88.2%) and young urologists (95%) were asked to follow a basic training or meet basic requirements before starting RAS, the requirements set by the educators were different from center to center. Some of them were required to attend only an online course on RAS, whereas others were asked to achieve threshold scores at VR simulator and participate in a standardized course at a training institute. The attendance to a structured advanced course in RAS showed a significant increase in surgical skills. Our study shows residents in urology are allowed to perform RAS during their residency though the criteria for starting RAS differ significantly amongst the teaching hospitals. To guarantee a basic level of skills and knowledge a structured, (multi-step) training and certification program for RAS should be implemented.


Risk factors for developing hepatocellular carcinoma in Egypt.
Hepatocellular carcinoma (HCC) is a common disorder worldwide and ranks 2nd and 6th most common cancer among men and women in Egypt. HCC has a rising incidence in Egypt mostly due to high prevalence of viral hepatitis and its complications. Proper management requires the interaction of multidisciplinary HCC clinic to choose the most appropriate plan. The different modalities of treatment include resection (surgery or transplantation), local ablation, chemoembolization, radioembolization and molecular targeted therapies. This paper summarizes both the environmental and host related risk factors of HCC in Egypt including well-established risk factors such as hepatitis virus infection, aflatoxin, as well as possible risk factors.


The unusually stable quaternary structure of human Cu,Zn-superoxide dismutase 1 is controlled by both metal occupancy and disulfide status.
The eukaryotic copper,zinc superoxide dismutases are remarkably stable dimeric proteins that maintain an intrasubunit disulfide bond in the reducing environment of the cytosol and are active under a variety of stringent denaturing conditions. The structural interplay of conserved disulfide bond and metal-site occupancy in human copper,zinc superoxide dismutase (hSOD1) is of increasing interest as these post-translational modifications are known to dramatically alter the catalytic chemistry, the subcellular localization, and the susceptibility of the protein to aggregation. Using biophysical methods, we find no significant change in the gross secondary or tertiary structure of the demetallated form upon reduction of the disulfide. Interestingly, reduction does lead to a dramatic change in the quaternary structure, decreasing the monomer-to-dimer equilibrium constant by at least four orders of magnitude. This reduced form of hSOD1 is monomeric, even at concentrations well above the physiological range. Either the addition of Zn(II) or the formation of the disulfide leads to a shift in equilibrium that favors the dimeric species, even at low protein concentrations (i.e. micromolar range). We conclude that only the most immature form of hSOD1, i.e. one without any post-translational modifications, favors the monomeric state under physiological conditions. This finding provides a basis for understanding the selectivity of mitochondrial SOD1 import and may be relevant to the toxic properties of mutant forms of hSOD1 that can cause the familial form of amyotrophic lateral sclerosis.


Inhibition of CD47 Effectively Targets Pancreatic Cancer Stem Cells via Dual Mechanisms.
Pancreatic ductal adenocarcinoma (PDAC) is a cancer of the exocrine pancreas with unmet medical need and is strongly promoted by tumor-associated macrophages (TAM). The presence of TAMs is associated with poor clinical outcome, and their overall role, therefore, appears to be protumorigenic. The "don't eat me" signal CD47 on cancer cells communicates to the signal regulatory protein-α on macrophages and prevents their phagocytosis. Thus, inhibition of CD47 may offer a new opportunity to turn TAMs against PDAC cells, including cancer stem cells (CSC), as the exclusively tumorigenic population. We studied in vitro and in vivo the effects of CD47 inhibition on CSCs using a large set of primary pancreatic cancer (stem) cells as well as xenografts of primary human PDAC tissue. CD47 was highly expressed on CSCs, but not on other nonmalignant cells in the pancreas. Targeting CD47 efficiently enhanced phagocytosis of a representative set of primary human pancreatic cancer (stem) cells and, even more intriguingly, also directly induced their apoptosis in the absence of macrophages during long-term inhibition of CD47. In patient-derived xenograft models, CD47 targeting alone did not result in relevant slowing of tumor growth, but the addition of gemcitabine or Abraxane resulted in sustained tumor regression and prevention of disease relapse long after discontinuation of treatment. These data are consistent with efficient in vivo targeting of CSCs, and strongly suggest that CD47 inhibition could be a novel adjuvant treatment strategy for PDAC independent of underlying and highly variable driver mutations.


Cdk1-dependent phosphorylation of Cdc13 coordinates telomere elongation during cell-cycle progression.
Elongation of telomeres by telomerase replenishes the loss of terminal telomeric DNA repeats during each cell cycle. In budding yeast, Cdc13 plays an essential role in telomere length homeostasis, partly through its interactions with both the telomerase complex and the competing Stn1-Ten1 complex. Previous studies in yeast have shown that telomere elongation by telomerase is cell cycle dependent, but the mechanism underlying this dependence is unclear. In S. cerevisiae, a single cyclin-dependent kinase Cdk1 (Cdc28) coordinates the serial events required for the cell division cycle, but no Cdk1 substrate has been identified among telomerase and telomere-associated factors. Here we show that Cdk1-dependent phosphorylation of Cdc13 is essential for efficient recruitment of the yeast telomerase complex to telomeres by favoring the interaction of Cdc13 with Est1 rather than the competing Stn1-Ten1 complex. These results provide a direct mechanistic link between coordination of telomere elongation and cell-cycle progression in vivo.


Studies on the active components and antioxidant activities of the extracts of Mimosa pudica Linn. from southern China.
The total flavonoid (TF) and total phenolic (TP) contents of the ethanol extracts of the whole plant, stem, leaf, and seed of Mimosa pudica Linn belonging to the genus Mimosa (Family: Fabaceae alt. Leguminosae), which originates from the subtropical regions of southern China, were determined in this experiment. The antioxidant activity of the extracts and 5 flavonoid monomers of M. pudica Linn. were also evaluated by 2 assays, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and ferric reducing/antioxidant power (FRAP) assays. In addition, correlation analysis was also made in the present study. The results showed that leaf extracts contained the highest amount of TF and TP, and the content was significantly higher than that found in other parts of the plant. Moreover, the sequence of antioxidant activity of the ethanol extracts was as follows: leaf > the whole plant > seed > stem; the sequence of the 5 flavonoid monomers was as follows: 5,7,3´,4´-tetrahydroxy-6-C-[β-D-apiose-(1→4)]-β-D-glycopyranosyl flavone (1) > isorientin (2) > orientin (3) > isovitexin (4) > vitexin (5), and the antioxidant activity of compound 1 is equivalent to the synthetic antioxidant trolox or a bit stronger than trolox, and significant correlations were found among the active ingredient contents and the results of antioxidant activity. The present study suggested that M. pudica Linn. could be a potential rich source of natural antioxidants.


The effect of vagal nerve blockade using electrical impulses on glucose metabolism in nondiabetic subjects.
Vagal interruption causes weight loss in humans and decreases endogenous glucose production in animals. However, it is unknown if this is due to a direct effect on glucose metabolism. We sought to determine if vagal blockade using electrical impulses alters glucose metabolism in humans. We utilized a randomized, cross-over study design where participants were studied after 2 weeks of activation or inactivation of vagal nerve blockade (VNB). Seven obese subjects with impaired fasting glucose previously enrolled in a long-term study to examine the effect of VNB on weight took part. We used a standardized triple-tracer mixed meal to enable measurement of the rate of meal appearance, endogenous glucose production, and glucose disappearance. The 550 kcal meal was also labeled with (111)In-diethylene triamine pentaacetic acid (DTPA) to measure gastrointestinal transit. Insulin action and β-cell responsivity indices were estimated using the minimal model. Integrated glucose, insulin, and glucagon concentrations did not differ between study days. This was also reflected in a lack of effect on β-cell responsivity and insulin action. Furthermore, fasting and postprandial endogenous glucose production, integrated meal appearance, and glucose disposal did not differ in the presence or absence of VNB. Similarly, gastric emptying and colonic transit were unchanged by VNB. In this pilot study in nondiabetic humans, electrical vagal blockade had no acute effects on glucose metabolism, insulin secretion and action, or gastric emptying. It remains to be determined if more pronounced effects would be observed in diabetic subjects.


Problems in measuring human leucocyte cation content: effects of cell preparation and handling.
1. Previous published measurements of leucocyte cation content are inconsistent, with sodium concentrations having particularly high coefficients of variation. We have measured the effects of recovery time after isolation, different types of handling, and centrifugation on leucocyte sodium and potassium content. 2. Sodium content fell markedly during the first 30 min after isolation and was stable from 1 to 3 h of incubation. There was a small but significant rise in potassium content over the same time period. 3. During incubation, occasional gentle resuspension of the cells gave optimal sodium contents, whereas mixing by inversion caused large falls in sodium and potassium content. 4. Centrifugation of stable cells at 200 g for 6 min caused marked increases in sodium content.


Alterations in myocardial cardiolipin content and composition occur at the very earliest stages of diabetes: a shotgun lipidomics study.
Recently, we have identified the dramatic depletion of cardiolipin (CL) in diabetic myocardium 6 weeks after streptozotocin (STZ) injection that was accompanied by increases in triacylglycerol content and multiple changes in polar lipid molecular species. However, after 6 weeks in the diabetic state, the predominant lipid hallmarks of diabetic cardiomyopathy were each present concomitantly, and thus, it was impossible to identify the temporal course of lipid alterations in diabetic myocardium. Using the newly developed enhanced shotgun lipidomics approach, we demonstrated the dramatic loss of abundant CL molecular species in STZ-treated hearts at the very earliest stages of diabetes accompanied by a profound remodeling of the remaining CL molecular species including a 16-fold increase in the content of 18:2-22:6-22:6-22:6 CL. These alterations in CL metabolism occur within days after the induction of the diabetic state and precede the triacylglycerol accumulation manifest in diabetic myocardium. Similarly, in ob/ob mice, a dramatic and progressive redistribution from 18:2 FA-containing CL molecular species to 22:6 FA-containing CL molecular species was also identified. Collectively, these results demonstrate alterations in CL hydrolysis and remodeling at the earliest stages of diabetes and are consistent with a role for alterations in CL content in precipitating mitochondrial dysfunction in diabetic cardiomyopathy.


Disease types discovery from a large database of inpatient records: A sepsis study.
Data-driven phenotype discoveries on Electronic Health Records (EHR) data have recently drawn benefits across many aspects of clinical practice. In the method described in this paper, we map a very large EHR database containing more than a million inpatient cases into a low dimensional space where diseases with similar phenotypes have similar representation. This embedding allows for an effective segmentation of diseases into more homogeneous categories, an important task of discovering disease types for precision medicine. In particular, many diseases have heterogeneous nature. For instance, sepsis, a systemic and progressive inflammation, can be caused by many factors, and can have multiple manifestations on different human organs. Understanding such heterogeneity of the disease can help in addressing many important issues regarding sepsis, including early diagnosis and treatment, which is of huge importance as sepsis is one of the main causes of in-hospital deaths in the United States. This study analyzes state of the art embedding models that have had huge success in various fields, applying them to disease embedding from EHR databases. Particular interest is given to learning multi-type representation of heterogeneous diseases, which leads to more homogeneous groups. Our results show evidence that such representations have phenotypes of higher quality and also provide benefit when predicting mortality of inpatient visits.


Requirement of Lyn and Syk tyrosine kinases for the prevention of apoptosis by cytokines in human eosinophils.
In allergic diseases, the cytokines interleukin (IL)5 and granulocyte/macrophage colony-stimulating factor (GM-CSF) are upregulated and have been proposed to cause blood and tissue eosinophilia by inhibition of eosinophil apoptosis. We demonstrate herein, in freshly isolated human eosinophils, that the IL-3/IL-5/GM-CSF receptor beta subunit interacts with cytoplasmic tyrosine kinases to induce phosphorylation of several cellular substrates, including the beta subunit itself. The Lyn and Syk intracellular tyrosine kinases constitutively associate at a low level with the IL-3/IL-5/GM-CSF receptor beta subunit in human eosinophils. Stimulation with GM-CSF or IL-5 results in a rapid and transient increase in the amount of Lyn and Syk associated with the IL-3/IL-5/GM-CSF receptor beta subunit. Lyn is required for optimal tyrosine phosphorylation and activation of Syk. In contrast, Syk is not required for optimal tyrosine phosphorylation and activation of Lyn. These data suggest that Lyn is proximal to Syk in a tyrosine kinase cascade that transduces IL-3, IL-5, or GM-CSF signals. Compatible with this model, both Lyn and Syk are essential for the activation of the antiapoptotic pathway(s) induced through the IL-3/IL-5/GM-CSF receptor beta subunit in human eosinophils.


Purification and properties of DNA polymerase from Bacillus caldotenax.
A thermostable DNA polymerase was prepared from Bacillus caldotenax by using a four-step chromatography procedure. The protein exists as a monomer of M(r) 94,000, has a pI of 4.9 and has no associated 3'-5' or 5'-3'-exonuclease activities or endonuclease activity. The temperature optimum of the enzyme was about 70 degrees C and the pH for maximum activity was about 7.5. The enzyme has an absolute requirement for a bivalent cation, and maximum activity was obtained at the unusually high concentration of 70 mM-MgCl2. Mg2+ could be replaced by MnCl2 or CoCl2, with decreased activity, at the lower optimal concentrations of 1 mM and 2.5 mM respectively. Enzyme activity was inhibited in the presence of 2',3'-dideoxy-TTP, arabinosyl-CTP and aphidicolin. Enzyme activity was stimulated with KCl concentrations of about 100 mM, and concentrations of univalent salts above about 150 mM inhibited activity. The enzyme could use activated calf thymus DNA, poly(dA).p(dT)10 or primed single-stranded phage M13 DNA as a template and maximum activity was obtained with poly(dA).p(dT)10. The enzyme was inactive on unprimed single-stranded DNA, double-stranded DNA and polyribonucleotide template/primer. The apparent Km values for individual dNTPs, determined with the other dNTPs at saturating concentrations, were 5.7 microM (dCTP), 6.3 microM (dATP, dGTP) and 6.4 microM (dTTP). The Km value for the overall incorporation of each dNTP from an equimolar mixture of all four dNTPs was 24.7 microM. The kcat. value was about 1.05 s-1. The kcat./Km value was 0.16-0.18 M-1.s-1 for individual dNTPs and 0.04 for the incorporation of an equimolar mixture of all four dNTPs. Some of the properties of the enzyme show it may be classified as an alpha-Type DNA polymerase.


A 33-year follow-up of narcotics addicts.
This study examined longitudinal patterns of heroin use, other substance use, health, mental health, employment, criminal involvement, and mortality among heroin addicts. The sample was composed of 581 male heroin addicts admitted to the California Civil Addict Program (CAP) during the years 1962 through 1964; CAP was a compulsory drug treatment program for heroin-dependent criminal offenders. This 33-year follow-up study updates information previously obtained from admission records and 2 face-to-face interviews conducted in 1974-1975 and 1985-1986; in 1996-1997, at the latest follow-up, 284 were dead and 242 were interviewed. In 1996-1997, the mean age of the 242 interviewed subjects was 57.4 years. Age, disability, years since first heroin use, and heavy alcohol use were significant correlates of mortality. Of the 242 interviewed subjects, 20.7% tested positive for heroin (with additional 9.5% urine refusal and 14.0% incarceration, for whom urinalyses were unavailable), 66.9% reported tobacco use, 22.1% were daily alcohol drinkers, and many reported illicit drug use (eg, past-year heroin use was 40.5%; marijuana, 35.5%; cocaine, 19.4%; crack, 10.3%; amphetamine, 11.6%). The group also reported high rates of health problems, mental health problems, and criminal justice system involvement. Long-term heroin abstinence was associated with less criminality, morbidity, psychological distress, and higher employment. While the number of deaths increased steadily over time, heroin use patterns were remarkably stable for the group as a whole. For some, heroin addiction has been a lifelong condition associated with severe health and social consequences.


Mechanism of aerobic biological destabilisation of wool scour effluent emulsions.
Wool scouring effluent is a highly polluted industrial wastewater in which the main pollutant, wool wax, is held in a stable oil-in-water emulsion by non-ionic detergent. The use of microbial action to cause emulsion destabilisation has been proposed as a new treatment strategy for this effluent stream. This strategy aims at improving aerobic treatment performance by physically removing the high-COD, slowly bio-degradable wool wax from the system without bio-degradation. The mechanism by which an aerobic-mixed culture destabilises the wool scouring effluent emulsion was investigated. Our results show that destabilisation is due to partial bio-degradation of both the scouring detergent and the wool wax. Cleavage of the wool wax esters was the first stage in wax degradation, when 40-50% of wax was de-emulsified. Over the same period, detergent degradation was low, at 7-21%. With further incubation, detergent degradation increased, aiding further breakdown of the emulsion. The degradation of the detergent, a nonylphenol ethoxylate, resulted in both a reduction in molar concentration (of up to 82%) and a shortening of the ethoxylate chain length. The latter reduced the hydrophile-lipophile balance (HLB) from 12 to approximately 7, thereby reducing the ability of the residual detergent to stabilise the emulsion. Analysis of the emulsified and de-emulsified wax fractions could not identify a group of compounds that were preferentially de-emulsified based on molecular weight or polarity. These findings will assist in using a de-emulsification strategy in both existing and new treatment systems in order to save on aeration costs and treatment times for biological treatment of this highly polluted wastewater.


Stimulated salivary flow rate and oral health status.
This study examined the relationship between stimulated salivary flow rate and oral health status in an adult population. Multinomial multivariate logistic regression analysis was used to examine the associations of salivary flow rate with dental caries status and periodontal status at the individual level among 2,110 Japanese adults with ≥10 teeth. Then, a spline model was used to examine the nonlinear relationship between salivary flow rate and teeth with dental caries or periodontal disease in multilevel analysis. Odds ratios were calculated for a 1.0-mL/min reduction in salivary flow rate at a point. After adjusting for confounding variables, participants with a flow rate ≤3.5 mL/min had significantly higher odds ratios for high caries status, and participants with a flow rate ≤1.4 mL/min had a higher odds ratio for broad periodontal disease, than did those with a flow rate >3.5 mL/min. In spline models, the odds ratio for teeth with dental caries or periodontal disease increased with reduced saliva secretion. The present findings suggest that decreased saliva secretion affects both dental caries and general periodontal health status.


Determinants of skilled birth attendant utilization in Afghanistan: a cross-sectional study.
We sought to identify characteristics associated with use of skilled birth attendants where health services exist in Afghanistan. We conducted a cross-sectional study in all 33 provinces in 2004, yielding data from 617 health facilities and 9917 women who lived near the facilities and had given birth in the past 2 years. Only 13% of respondents had used skilled birth attendants. Women from the wealthiest quintile (vs the poorest quintile) had higher odds of use (odds ratio [OR] = 6.3; 95% confidence interval [CI] = 4.4, 8.9). Literacy was strongly associated with use (OR = 2.5; 95% CI = 2.0, 3.2), as was living less than 60 minutes from the facility (OR = 1.5; 95% CI = 1.1, 2.0) and residing near a facility with a female midwife or doctor (OR = 1.4; 95% CI = 1.1, 1.8). Women living near facilities that charged user fees (OR = 0.8; 95% CI = 0.6, 1.0) and that had male community health workers (OR = 0.6; 95% CI = 0.5, 0.9) had lower odds of use. In Afghanistan, the rate of use of safe delivery care must be improved. The financial barriers of poor and uneducated women should be reduced and culturally acceptable alternatives must be considered.


Cytochrome P450 expression and activities in rat, rabbit and bovine tongue.
Xenobiotic metabolism in the tongue has received little attention in the literature. In the present study, we report a comparative analysis of constitutive cytochrome P450 (CYP) expression and activities in the tongue. First we compared catalytic activities of rabbit, rat and bovine tongue samples using the probe substrates 4-nitrophenol, 1-phenylethanol, caffeine and 7-ethoxycoumarin. Rabbit tongue samples showed the highest activities for all substrates. We then compared the activities in rat and rabbit tongue with those in the rabbit liver, along with the effects of P450 inhibitors on specific activities. Combined, the activity studies indicate that CYP1A1 is active in rabbit tongue cells, but CYP1A2, CYP3A6 and CYP2E1 are below limits of detection. RT-PCR was also used to compare mRNA levels of 11 different rabbit and six different rat P450 isoforms in the tongue to those in the liver of these two species. Only CYP2E1, CYP1A1 and CYP4A4 were detected at significant levels in the rabbit tongue. None of the six rat isoforms probed were observed in the tongue. Although 4-nitrophenol activity was observed in the rabbit tongue samples, the kinetic parameter K(m) was inconsistent with the involvement of CYP2E1. We suggest that although CYP2E1 is expressed in the tongue, it is rapidly degraded in this organ, and the nitrophenol hydroxylation and caffeine hydroxylation we observe is the result of activity of CYP1A1.


The practices of psychologists working in schools during COVID-19: A multi-country investigation.
This exploratory study aimed to identify the ways psychologists working in schools supported students' mental health during school closures related to the COVID-19 pandemic. An online survey was developed to determine (a) how psychologists working in schools across the United States, Canada, Germany, and Australia supported students' mental health during COVID-19, (b) how their services changed during COVID-19, and (c) potential differences between countries concerning difficulties supporting students' mental health during this time. The survey was based on previous research and was subsequently piloted. Using convenience and snowball sampling, 938 participants (U.S. n = 665; Canada n = 48; Germany n = 140; Australia n = 85) completed the online survey. Overall, school psychology services across these four countries pivoted from psychoeducational assessments to virtual counseling, consultation, and the development/posting of online support directly to children or parents to use with their children. There was some variation between countries; during the pandemic, significantly more psychologists in Germany and Australia provided telehealth/telecounseling than those in the United States and Canada, and psychologists in Germany provided significantly more hardcopy material to support children than psychologists in other countries. There is a need to ensure psychologists have the appropriate technological skills to support school communities during periods of school closure, including, but not limited to, virtual counseling and the administration of psychoeducational assessments. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


MET signaling in colon cancer stem-like cells blunts the therapeutic response to EGFR inhibitors.
Metastatic colorectal cancer remains largely incurable, although in a subset of patients, survival is prolonged by new targeting agents such as anti-EGF receptor (anti-EGFR) antibodies. This disease is believed to be supported by a subpopulation of stem-like cells termed colon cancer-initiating cell (CCIC), which may also confer therapeutic resistance. However, how CCICs respond to EGFR inhibition has not been fully characterized. To explore this question, we systematically generated CCICs through spheroid cultures of patient-derived xenografts of metastatic colorectal cancer. These cultures, termed "xenospheres," were capable of long-term self-propagation in vitro and phenocopied the original patient tumors in vivo, thus operationally defining CCICs. Xenosphere CCICs retained the genetic determinants for EGFR therapeutic response in vitro and in xenografts; like the original tumors, xenospheres harboring a mutated KRAS gene were resistant to EGFR therapy, whereas those harboring wild-type RAS pathway genes (RAS(wt)) were sensitive. Notably, the effects of EGFR inhibition in sensitive CCICs could be counteracted by cytokines secreted by cancer-associated fibroblasts. In particular, we found that the MET receptor ligand hepatocyte growth factor (HGF) was especially active in supporting in vitro CCIC proliferation and resistance to EGFR inhibition. Ectopic production of human HGF in CCIC xenografts rendered the xenografts susceptible to MET inhibition, which sensitized the response to EGFR therapy. By showing that RAS(wt) CCICs rely on both EGFR and MET signaling, our results offer a strong preclinical proof-of-concept for concurrent targeting of these two pathways in the clinical setting.


Lipopolysaccharide induces pro-inflammatory cytokines and MMP production via TLR4 in nasal polyp-derived fibroblast and organ culture.
Nasal polyposis is characterized by persistent inflammation and remodeling in sinonasal mucosa. Toll-like receptors (TLRs) play a role in the innate immune response to microbes in the sinonasal cavity. The aim of this study was to evaluate whether nasal polyp-derived fibroblasts (NPDFs) and organ-cultured nasal polyps can synthesize pro-inflammatory cytokines and matrix metalloproteinases (MMPs) after exposure to lipopolysaccharide (LPS), a TLR4 agonist. NPDFs and organ-cultured nasal polyps were isolated from nasal polyps of 8 patients and exposed to LPS. The mRNA and protein expression levels of TLRs, cytokines, and MMPs were determined using a gene expression microarray, real-time RT-PCR, western blot analysis, enzyme-linked immunosorbent assay, and immunofluorescence staining. The enzymatic activities of MMPs were analyzed using collagen or gelatin zymography. The protein expression level of MMP-1 increased in nasal polyp tissues compared to inferior turbinate tissues. LPS induced mRNA expression of TLR4, IL-6, IL-8, and MMP-1 and activated MAPK signaling in NPDFs. LPS promoted the release of interleukin (IL)-6 through extracellular signal-related kinase (ERK) and IL-8 through ERK and c-Jun N-terminal kinases (JNK). Production of IL-6 and IL-8 was induced by PI3K/Akt signaling in LPS-stimulated NPDFs. LPS increased the transcript and protein expression levels of MMP-1 and induced collagenase activity of MMP-1 via ERK and p38, but did not induce gelatinase activity of MMP-2 and MMP-9. LPS from Rhodobacter sphaeroides (LPS-RS) inhibited the stimulatory effects of LPS in NPDFs as well as in organ culture of nasal polyp. LPS triggers immune response via TLR 4 and activates MAPK and PI3K/Akt signaling pathway, which is involved in remodeling of nasal polyps.


Metabolic recovery of Arabidopsis thaliana roots following cessation of oxidative stress.
To cope with the various environmental stresses resulting in reactive oxygen species (ROS) production plant metabolism is known to be altered specifically under different stresses. After overcoming the stress the metabolism should be reconfigured to recover basal operation however knowledge concerning how this is achieved is cursory. To investigate the metabolic recovery of roots following oxidative stress, changes in metabolite abundance and carbon flow were analysed. Arabidopsis roots were treated by menadione to elicit oxidative stress. Roots were fed with (13)C labelled glucose and the redistribution of isotope was determined in order to study carbon flow. The label redistribution through many pathways such as glycolysis, the tricarboxylic acid (TCA) cycle and amino acid metabolism were reduced under oxidative stress. After menadione removal many of the stress-related changes reverted back to basal levels. Decreases in amounts of hexose phosphates, malate, 2-oxoglutarate, glutamate and aspartate were fully recovered or even increased to above the control level. However, some metabolites such as pentose phosphates and citrate did not recover but maintained their levels or even increased further. The alteration in label redistribution largely correlated with that in metabolite abundance. Glycolytic carbon flow reverted to the control level only 18 h after menadione removal although the TCA cycle and some amino acids such as aspartate and glutamate took longer to recover. Taken together, plant root metabolism was demonstrated to be able to overcome menadione-induced oxidative stress with the differential time period required by independent pathways suggestive of the involvement of pathway specific regulatory processes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-011-0296-1) contains supplementary material, which is available to authorized users.


Outcomes of Posterior Glenoid Bone-Grafting in Anatomical Total Shoulder Arthroplasty: A Systematic Review.
Total shoulder arthroplasty offers a reliable means with which to treat glenohumeral joint arthritis. Posterior glenoid bone loss presents a unique challenge with an increased risk of glenoid component failure. The use of posterior bone-grafting is one method to address glenoid bone loss in patients undergoing anatomical total shoulder arthroplasty. The purpose of the present study was to assess the outcome and survival of the glenoid component following the use of bone graft to address posterior glenoid bone loss in patients undergoing anatomical total shoulder arthroplasty. A systematic review of posterior glenoid bone-grafting in patients undergoing anatomical total shoulder arthroplasty was performed. Studies evaluating patient-reported outcomes, complications, and imaging assessments of the glenoid component as well as of bone graft structural healing and integrity following posterior glenoid bone-grafting were included. Data extracted included demographic characteristics, Walch classification, bone-grafting method, clinical outcomes measures, complications, radiolucency around the glenoid component, graft failure, posterior humeral head subluxation, and time of the latest follow-up. Six studies met the inclusion and exclusion criteria. Ninety-four patients from these 6 studies underwent posterior glenoid bone-grafting with anatomical total shoulder arthroplasty. The mean age was 59.7 years, and the mean duration of follow-up was 5.7 years. Overall, 57% of the patients had an excellent postoperative Neer score. At the time of the latest follow-up, 28.7% had evidence of radiolucency and 35% had humeral head subluxation or instability. Thirteen patients (14%) underwent revision of the glenoid component by the time of the latest follow-up. The present study demonstrated a 28.7% complication rate, a 14% revision rate, a 17% graft failure rate, and a 35% rate of recurrence of posterior humeral head subluxation. Posterior glenoid bone-grafting to correct bone loss is associated with a substantial risk of postoperative complications. The treatment of posterior glenoid bone loss remains a challenge in patients undergoing anatomical total shoulder arthroplasty. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Functional analysis of ectopic parathyroid activity in cats.
The ability of ectopic parathyroid tissue to support calcium homeostasis was evaluated by measuring serum concentrations of calcium, phosphorus, albumin, magnesium, and parathyroid hormone before and for 12 weeks after bilateral thyroparathyroidectomy in 14 cats. During the immediate postoperative period, significant decrease was observed in serum calcium, magnesium, and parathyroid hormone (PTH) concentrations. Serum PTH concentration remained subnormal and did not significantly increase during the 12-week observation period. Despite persistent hypoparathyroidism, serum calcium and magnesium concentrations gradually increased. Ectopic parathyroid tissue is not capable of maintaining normal serum calcium concentration immediately after thyroparathyroidectomy. Serum calcium concentration gradually normalizes after thyroparathyroidectomy, apparently by means of a PTH-independent mechanism.