Biotransformation of Ilaprazole in Human Liver Microsomes and Human: Role of CYP3A4 in Ilaprazole Clearance and Drug-Drug Interaction.
Ilaprazole is a new proton pump inhibitor and is currently marketed in China and South Korea for the treatment of gastric and duodenal ulcer. Ilaprazole has a favorable long half-life and minimal pharmacokinetic variability associated with CYP2C19 polymorphism. Sulfoxide oxidation of ilaprazole is catalyzed mainly by CYP3A4. Thus, it has been widely accepted that CYP3A4 plays a major role in the clearance of ilaprazole in humans. However, absorption, distribution, metabolism, and excretion data of radiolabeled ilaprazole in humans are not available. The primary goal of this study was to determine if sulfoxide oxidation is a major metabolic pathway of ilaprazole in humans. Metabolite profiles of ilaprazole, ilaprazole sulfide, and ilaprazole sulfone in human liver microsomes (HLMs) were characterized and quantitively analyzed by liquid chromatography (LC)/UV/high-resolution mass spectrometry (HRMS). Moreover, metabolites of ilaprazole in human urine and feces were detected and identified by LC-HRMS. The results revealed that sulfoxide reduction to ilaprazole sulfide rather than sulfoxide oxidation was the major biotransformation pathway in HLMs. Sulfoxide reduction also occurred in HLMs without NADPH or in deactivated HLMs. Ilaprazole sulfide and its multiple oxidative metabolites were major drug-related components in human urine and feces, where there were no ilaprazole sulfone and its metabolites. A small amount of the parent drug was found in feces. Thus, we propose that nonenzymatic sulfoxide reduction rather than CYP3A4-medidated sulfoxide oxidation is the major metabolic clearance pathway of ilaprazole in humans. Consequently, it is predicted that ilaprazole has no significant drug-drug interaction via CYP3A4 inhibition or induction by a coadministered drug.


Cognitive appraisals and physical health in people with posttraumatic stress disorder (PTSD).
Previous literature suggests that posttraumatic stress disorder (PSTD) is associated with poor physical health. To date a number of existing hypotheses have been developed to explain this association focusing predominantly on the interplay between cognitive and emotional appraisals. We have attempted to synthesise existing literature on the role of cognitive and emotional appraisals in explaining ill health (i.e. medically explained symptoms), following PTSD. On the basis of this review, we are introducing a conceptualisation aiming to explain poor physical health following PTSD. This new conceptualisation proposes that PTSD symptoms will lead to the production of two types of cognitive appraisals: Appraisals about the body that occurred during traumatisation, and cognitive appraisals about PTSD symptoms themselves. Both these cognitive appraisals coupled with negative emotional responses will lead to physiological arousal and negative health behaviours and subsequently to poor health. Although this conceptualisation has certain clinical implications for the management of physical health following PTSD, further research is required to test such by investigating the association between its elements.


Two transcription products of the vesicular stomatitis virus genome may control L-cell protein synthesis.
When mouse L-cells are infected with vesicular stomatitis virus, there is a decrease in the rate of protein synthesis ranging from 20 to 85% of that in mock-infected cells. Vesicular stomatitis virus, irradiated with increasing doses of UV light, eventually loses this capacity to inhibit protein synthesis. The UV inactivation curve was biphasic, suggesting that transcription of two regions of the viral genome is necessary for the virus to become inactivated in this capacity. The first transcription product corresponded to about 373 nucleotides, and the second corresponded to about 42 nucleotides. Inhibition of transcription of the larger product by irradiating the virus with low doses of UV light left a residual inhibition of protein synthesis consisting of approximately 60 to 65% of the total inhibition. This residual inhibition could be obviated by irradiating the virus with a UV dose of greater than 20,000 ergs/mm(2) and was thus considered to represent the effect of the smaller transcription product. In the R1 mutant of C. P. Stanners et al. (Cell 11:273-281, 1977), inhibition of transcription of the larger product sufficed to restore protein synthesis to the mock-infected level, suggesting that the smaller transcription product is nonfunctional with respect to protein synthesis inhibition. It thus appears that the inhibition of protein synthesis by wild-type vesicular stomatitis virus involved at least two separate viral transcription products, and the inhibition by the R1 mutant involved only one. Extracts from cells infected with virus irradiated with low doses of UV light showed a protein synthesis capacity quite similar to that of their in vivo counterparts, indicating that these extracts closely reflect the in vivo effects of virus infection.


Treating gynaecological disorders with traditional Chinese medicine: a review.
Traditional Chinese Medicine (TCM) has significant advantages in treating gynaecological disorders. The paper has provided a brief introduction on the current progress of treating some gynaecological disorders including endometriosis, infertility, dysmenorrhea, abnormal uterine bleeding, premenstrual syndrome, menopausal syndrome, uterine fibroids, chronic pelvic inflammation, polycystic ovarian syndrome (PCOS), cervicitis and vaginitis with Chinese Herbal Medicine (CHM) and acupuncture. The use of TCM in the field of assisted reproductive techniques (ART) has also been included in the review. In addition, thirty-two commonly used Chinese medicinal formulas in treating gynaecological disorders have been introduced.


Colchicine is a competitive antagonist at human recombinant gamma-aminobutyric acidA receptors.
Colchicine is an alkaloid that is used clinically in the treatment of arthritic gout. This potent microtubule disrupting agent has also been used extensively as an experimental tool in studies characterizing the role of the cytoskeleton in a variety of cellular processes. Colchicine has also been used as a selective neurotoxin and in animal models of Alzheimer's disease and epilepsy. Although the mechanism(s) mediating the neurotoxic actions of colchicine have not been established, most studies have attributed these effects to its microtubule depolymerizing actions. Here we report another central nervous system action of colchicine, competitive antagonism of gamma-aminobutyric acid (GABA)A receptor function. By use of a rapid drug perfusion system, colchicine (10-1000 microM) significantly inhibited GABA currents recorded from L(tk-) cells stably transfected with human alpha 1 beta 2 gamma 2L GABAA receptor subunits. The inhibition was rapid and reversible, with 100 microM colchicine shifting the GABA EC50 from 2.5 to 5.1 microM with no effect on currents evoked by saturating concentrations of GABA. Colchicine also significantly inhibited binding of the competitive GABAA receptor antagonist [3H]SR-95531. Other microtubule disrupting agents (10 microM vinblastine, 10 micrograms/ml nocodazole, 1 microM taxol) had no acute effects on GABA currents, nor did the inactive analog gamma-lumicolchicine (100 microM). Moreover, pretreating cells with colchicine, vinblastine, nocodazole or taxol for 1 to 4 hr did not occlude the acute inhibitory action of colchicine. We conclude that, in addition to its well characterized effects on microtubule assembly, colchicine can also inhibit GABAA receptor function through a direct interaction with the receptor/ion channel complex.


Delineating the conformational flexibility of trisaccharides from NMR spectroscopy experiments and computer simulations.
The conformation of saccharides in solution is challenging to characterize in the context of a single well-defined three-dimensional structure. Instead, they are better represented by an ensemble of conformations associated with their structural diversity and flexibility. In this study, we delineate the conformational heterogeneity of five trisaccharides via a combination of experimental and computational techniques. Experimental NMR measurements target conformationally sensitive parameters, including J couplings and effective distances around the glycosidic linkages, while the computational simulations apply the well-calibrated additive CHARMM carbohydrate force field in combination with efficient enhanced sampling molecular dynamics simulation methods. Analysis of conformational heterogeneity is performed based on sampling of discreet states as defined by dihedral angles, on root-mean-square differences of Cartesian coordinates and on the extent of volume sampled. Conformational clustering, based on the glycosidic linkage dihedral angles, shows that accounting for the full range of sampled conformations is required to reproduce the experimental data, emphasizing the utility of the molecular simulations in obtaining an atomic detailed description of the conformational properties of the saccharides. Results show the presence of differential conformational preferences as a function of primary sequence and glycosidic linkage types. Significant differences in conformational ensembles associated with the anomeric configuration of a single glycosidic linkage reinforce the impact of such changes on the conformational properties of carbohydrates. The present structural insights of the studied trisaccharides represent a foundation for understanding the range of conformations adopted in larger oligosaccharides and how these molecules encode their conformational heterogeneity into the monosaccharide sequence.


Expression of cyclin D2 and D3 in lymphoid lesions.
The D-type cyclins, involved in the regulation of G1 progression of the cell cycle, are expressed in a lineage-specific manner. Normal hematopoietic cells express cyclin D2 and/or D3. In order to determine whether their expression pattern changes in lymphoid tumors, we examined cyclin D2 and D3 expression in non-neoplastic and neoplastic lymphoid lesions, using a sensitive immunohistochemical amplification method. Centroblasts in lymphoid follicles of reactive lymph nodes expressed exclusively cyclin D3 and no D2. Interfollicular areas contained scattered cyclin D3 and D2 positive cells. By double staining, cyclin D3 was detected in CD79a positive B cells, CD3 positive T cells and CD68 positive macrophages. Cyclin D2 was present only in CD3 positive T cells. Neoplastic lymphoid lesions included 33 B cell lymphomas, 9 T cell lymphomas and 12 Hodgkin's lymphomas. The B cell lymphomas comprised 9 follicular lymphomas (FL), 1 Burkitt lymphoma (BL), 22 diffuse large cell lymphomas (DL) and 1 chronic lymphocytic leukemia (CLL). All 9 FLs and the single BL expressed exclusively cyclin D3, similarly to germinal center B cells, that represent their cells of origin. Six DLs expressed both cyclin D2 and D3, while 6 expressed only D3. Among the 9 pleomorphic T cell lymphomas, medium and large cell type, 5 expressed cyclin D2. Cyclin D3 was also detected in scattered cells in 4 of 9 cases and was highly expressed in 2 of 9 T cell lymphomas. The majority of Hodgkin's lymphomas expressed both cyclin D2 and D3 in Hodgkin/Reed-Sternberg (HRS) cells. The high frequency of positive cells indicates that both cyclins were expressed in the same cells.


Africanized bees extend their distribution in California.
Africanized honey bees (Apis mellifera) arrived in the western hemisphere in the 1950s and quickly spread north reaching California in the 1990s. These bees are highly defensive and somewhat more difficult to manage for commercial purposes than the European honey bees traditionally kept. The arrival of these bees and their potentially replacing European bees over much of the state is thus of great concern. After a 25 year period of little systematic sampling, a recent small scale study found Africanized honey bees in the Bay Area of California, far north of their last recorded distribution. The purpose of the present study was to expand this study by conducting more intensive sampling of bees from across northern California. We found Africanized honey bees as far north as Napa and Sacramento. We also found Africanized bees in all counties south of these counties. Africanized honey bees were particularly abundant in parts of the central valley and Monterey. This work suggests the northern spread of Africanized honey bees may not have stopped. They may still be moving north at a slow rate, although due to the long gaps in sampling it is currently impossible to tell for certain. Future work should routinely monitor the distribution of these bees to distinguish between these two possibilities.


Sex-specific risk of cardiovascular disease and cognitive decline: pregnancy and menopause.
Understanding the biology of sex differences is integral to personalized medicine. Cardiovascular disease and cognitive decline are two related conditions, with distinct sex differences in morbidity and clinical manifestations, response to treatments, and mortality. Although mortality from all-cause cardiovascular diseases has declined in women over the past five years, due in part to increased educational campaigns regarding the recognition of symptoms and application of treatment guidelines, the mortality in women still exceeds that of men. The physiological basis for these differences requires further research, with particular attention to two physiological conditions which are unique to women and associated with hormonal changes: pregnancy and menopause. Both conditions have the potential to impact life-long cardiovascular risk, including cerebrovascular function and cognition in women. This review draws on epidemiological, translational, clinical, and basic science studies to assess the impact of hypertensive pregnancy disorders on cardiovascular disease and cognitive function later in life, and examines the effects of post-menopausal hormone treatments on cardiovascular risk and cognition in midlife women. We suggest that hypertensive pregnancy disorders and menopause activate vascular components, i.e., vascular endothelium and blood elements, including platelets and leukocytes, to release cell-membrane derived microvesicles that are potential mediators of changes in cerebral blood flow, and may ultimately affect cognition in women as they age. Research into specific sex differences for these disease processes with attention to an individual's sex chromosomal complement and hormonal status is important and timely.


Uptake and translocation of Ti from nanoparticles in crops and wetland plants.
Bioavailability of engineered metal nanoparticles affects uptake in plants, impacts on ecosystems, and phytoremediation. We studied uptake and translocation of Ti in plants when the main source of this metal was TiO2 nanoparticles. Two crops (Phaseolus vulgaris (bean) and Triticum aestivum (wheat)), a wetland species (Rumex crispus, curly dock), and the floating aquatic plant (Elodea canadensis, Canadian waterweed), were grown in nutrient solutions with TiO2 nanoparticles (0, 6, 18 mmol Ti L(-1) for P. vulgaris, T. aestivum, and R. crispus; and 0 and 12 mmol Ti L(-1) for E. canadensis). Also examined in E. canadensis was the influence of TiO2 nanoparticles upon the uptake of Fe, Mn, and Mg, and the influence of P on Ti uptake. For the rooted plants, exposure to TiO2 nanoparticles did not affect biomass production, but significantly increased root Ti sorption and uptake. R. crispus showed translocation of Ti into the shoots. E. canadensis also showed significant uptake of Ti, P in the nutrient solution significantly decreased Ti uptake, and the uptake patterns of Mn and Mg were altered. Ti from nano-Ti was bioavailable to plants, thus showing the potential for cycling in ecosystems and for phytoremediation, particularly where water is the main carrier.


Diagnosis and treatment of Clostridium difficile in adults: a systematic review.
Since 2000, the incidence and severity of Clostridium difficile infection (CDI) have increased. To review current evidence regarding best practices for the diagnosis and treatment of CDI in adults (age ≥ 18 years). Ovid MEDLINE and Cochrane databases were searched using keywords relevant to the diagnosis and treatment of CDI in adults. Articles published between January 1978 and October 31, 2014, were selected for inclusion based on targeted keyword searches, manual review of bibliographies, and whether the article was a guideline, systematic review, or meta-analysis published within the past 10 years. Of 4682 articles initially identified, 196 were selected for full review. Of these, the most pertinent 116 articles were included. Clinical trials, large observational studies, and more recently published articles were prioritized in the selection process. Laboratory testing cannot distinguish between asymptomatic colonization and symptomatic infection with C difficile. Diagnostic approaches are complex due to the availability of multiple testing strategies. Multistep algorithms using polymerase chain reaction (PCR) for the toxin gene(s) or single-step PCR on liquid stool samples have the best test performance characteristics (for multistep: sensitivity was 0.68-1.00 and specificity was 0.92-1.00; and for single step: sensitivity was 0.86-0.92 and specificity was 0.94-0.97). Vancomycin and metronidazole are first-line therapies for most patients, although treatment failures have been associated with metronidazole in severe or complicated cases of CDI. Recent data demonstrate clinical success rates of 66.3% for metronidazole vs 78.5% for vancomycin for severe CDI. Newer therapies show promising results, including fidaxomicin (similar clinical cure rates to vancomycin, with lower recurrence rates for fidaxomicin, 15.4% vs vancomycin, 25.3%; P = .005) and fecal microbiota transplantation (response rates of 83%-94% for recurrent CDI). Diagnostic testing for CDI should be performed only in symptomatic patients. Treatment strategies should be based on disease severity, history of prior CDI, and the individual patient's risk of recurrence. Vancomycin is the treatment of choice for severe or complicated CDI, with or without adjunctive therapies. Metronidazole is appropriate for mild disease. Fidaxomicin is a therapeutic option for patients with recurrent CDI or a high risk of recurrence. Fecal microbiota transplantation is associated with symptom resolution of recurrent CDI but its role in primary and severe CDI is not established.


Interaction analysis of the CBLB and CTLA4 genes in type 1 diabetes.
Gene-gene interaction analyses have been suggested as a potential strategy to help identify common disease susceptibility genes. Recently, evidence of a statistical interaction between polymorphisms in two negative immunoregulatory genes, CBLB and CTLA4, has been reported in type 1 diabetes (T1D). This study, in 480 Danish families, reported an association between T1D and a synonymous coding SNP in exon 12 of the CBLB gene (rs3772534 G>A; minor allele frequency, MAF=0.24; derived relative risk, RR for G allele=1.78; P=0.046). Furthermore, evidence of a statistical interaction with the known T1D susceptibility-associated CTLA4 polymorphism rs3087243 (laboratory name CT60, G>A) was reported (P<0.0001), such that the CBLB SNP rs3772534 G allele was overtransmitted to offspring with the CTLA4 rs3087243 G/G genotype. We have, therefore, attempted to obtain additional support for this finding in both large family and case-control collections. In a primary analysis, no evidence for an association of the CBLB SNP rs3772534 with disease was found in either sample set (2162 parent-child trios, P=0.33; 3453 cases and 3655 controls, P=0.69). In the case-only statistical interaction analysis between rs3772534 and rs3087243, there was also no support for an effect (1994 T1D affected offspring, and 3215 cases, P=0.92). These data highlight the need for large, well-characterized populations, offering the possibility of obtaining additional support for initial observations owing to the low prior probability of identifying reproducible evidence of gene-gene interactions in the analysis of common disease-associated variants in human populations.


Oral probiotics influence oral and respiratory tract infections in pediatric population: a randomized double-blinded placebo-controlled pilot study.
Acute oral and respiratory tract infections (RTIs) are highly present in the general population, and they represent one of the most impacting causes of morbidity and mortality every year. The aim of our study was to assess the clinical impact of oral probiotics on acute oral and respiratory tract infections affecting paediatric patients. This is a randomized, double-blinded placebo-controlled clinical study, where probiotics have been compared with placebo in a double-blinded investigation. 40 subjects with a recent clinical history reporting oral and respiratory tract infections were randomly selected and assigned to control (n=20) or the probiotics (n=20) group. During the 3 months before the starting, all subjects were assessed with several salivary examinations. We found significant differences in the incidence of oral and respiratory tract infections between control and probiotic group, with a marked reduction of oral and respiratory tract infections episodes in the probiotic group. Salivary examinations gave similar results in both the groups, however, despite the salivary flow, viscosity and buffering were similar, on the contrary, pH values were found to be more alkaline in patients of probiotic group. This study showed the main positive effects related to a supplementation with probiotics in order to prevent or reduce the incidence of infections onset in oral and respiratory tracts without any drugs-related adverse effects.


Structural MRI markers of brain aging early after ischemic stroke.
To examine associations between ischemic stroke, vascular risk factors, and MRI markers of brain aging. Eighty-one patients (mean age 67.5 ± 13.1 years, 31 left-sided, 61 men) with confirmed first-ever (n = 66) or recurrent (n = 15) ischemic stroke underwent 3T MRI scanning within 6 weeks of symptom onset (mean 26 ± 9 days). Age-matched controls (n = 40) completed identical testing. Multivariate regression analyses examined associations between group membership and MRI markers of brain aging (cortical thickness, total brain volume, white matter hyperintensity [WMH] volume, hippocampal volume), normalized against intracranial volume, and the effects of vascular risk factors on these relationships. First-ever stroke was associated with smaller hippocampal volume (p = 0.025) and greater WMH volume (p = 0.004) relative to controls. Recurrent stroke was in turn associated with smaller hippocampal volume relative to both first-ever stroke (p = 0.017) and controls (p = 0.001). These associations remained significant after adjustment for age, sex, education, and, in stroke patients, infarct volume. Total brain volume was not significantly smaller in first-ever stroke patients than in controls (p = 0.056), but the association became significant after further adjustment for atrial fibrillation (p = 0.036). Cortical thickness and brain volumes did not differ as a function of stroke type, infarct volume, or etiology. Brain structure is likely to be compromised before ischemic stroke by vascular risk factors. Smaller hippocampal and total brain volumes and increased WMH load represent proxies for underlying vascular brain injury.


Tissue Engineering Auricular Cartilage Using Late Passage Human Auricular Chondrocytes.
The significant shortcomings associated with current autologous reconstructive options for auricular deformities have inspired great interest in a tissue engineering solution. A major obstacle in the engineering of human auricular cartilage is the availability of sufficient autologous human chondrocytes. A clinically obtainable amount of auricular cartilage tissue (ie, 1 g) only yields approximately 10 million cells, where 25 times this amount is needed for the fabrication of a full-scale pediatric ear. It is thought that repeated passaging of chondrocytes leads to dedifferentiation and loss of the chondrogenic potential. However, little to no data exist regarding the ideal number of times that human auricular chondrocytes (HAuCs) can be passaged in a manner that maximizes the cellular expansion while minimizing dedifferentiation. Human auricular chondrocytes were isolated from discarded otoplasty specimens. The HAuCs were then expanded, and cells from passages 3, 4, and 5 were encapsulated into discs 8 mm in diameter made from type I collagen hydrogels with a cell density of 25 million cells/mL. The constructs were implanted subcutaneously in the dorsa of nude mice and harvested after 1 and 3 months for analysis. Constructs containing passages 3, 4, and 5 chondrocytes all maintained their original cylindrical geometry. After 3 months in vivo, the diameters of the P3, P4, and P5 discs were 69 ± 9%, 67 ± 10%, and 73 ± 15% of their initial diameter, respectively. Regardless of the passage number, all constructs developed a glossy white cartilaginous appearance, similar to native auricular cartilage. Histologic analysis demonstrated development of an organized perichondrium composed of collagen, a rich proteoglycan matrix, cellular lacunae, and a dense elastin fibrin network by Safranin-O and Verhoeff stain. Biochemical analysis confirmed similar amounts of proteoglycan and hydroxyproline content in late passage constructs when compared with native auricular cartilage. These data indicate that late passage HAuCs (up to passage 5) form elastic cartilage that is histologically, biochemically, and biomechanically similar to native human elastic cartilage and have the potential to be used for auricular cartilage engineering.


Composition and applications of Aloe vera leaf gel.
Many of the health benefits associated with Aloe vera have been attributed to the polysaccharides contained in the gel of the leaves. These biological activities include promotion of wound healing, antifungal activity, hypoglycemic or antidiabetic effects antiinflammatory, anticancer, immunomodulatory and gastroprotective properties. While the known biological activities of A. vera will be briefly discussed, it is the aim of this review to further highlight recently discovered effects and applications of the leaf gel. These effects include the potential of whole leaf or inner fillet gel liquid preparations of A. vera to enhance the intestinal absorption and bioavailability of co-administered compounds as well as enhancement of skin permeation. In addition, important pharmaceutical applications such as the use of the dried A. vera gel powder as an excipient in sustained release pharmaceutical dosage forms will be outlined.


Behavioral responses to DDT and pyrethroids between Anopheles minimus species A and C, malaria vectors in Thailand.
Behavioral responses of two field populations of Anopheles minimus complex species A and C for contact and non-contact actions of chemicals were compared during and after exposure to operational field concentrations of DDT (2 g/m2), deltamethrin (0.02 g/m2), and lambdacyhalothrin (0.03 g/m2) using an excito-repellency escape chamber. The two populations were collected from the Mae Sot District in Tak Province (species A) and the Tri Yok District in Kanchanaburi Province (species C) in western Thailand. Female mosquitoes of both populations rapidly escaped from chambers after direct contact with DDT, deltamethrin, and lambdacyhalothrin. The non-contact repellency response to DDT and the two synthetic pyrethroids was pronounced with An. minimus species A; however, non-contact repellency was relatively weak with An. minimus species C, but remained significantly greater than the paired controls (P < 0.05). We conclude that strong contact irritancy was present in both test populations, whereas non-contact repellency also played a significant role in the escape response of An. minimus species A.


TMD in children and adolescents: prevalence of pain, gender differences, and perceived treatment need.
The aim of this study was to determine the prevalence of pain that is related to temporomandibular disorders (TMD), gender differences, and perceived treatment need in children and adolescents at a public dental clinic in Linköping, Sweden. A total of 862 children and adolescents aged 12 to 18 years received a questionnaire and their jaw opening was measured. Those who reported pain once a week or more in the masticatory system received a more comprehensive examination, including the Research Diagnostic Criteria for TMD and a neurologic examination (group 1). Group 2 reported pain less than once a week. Seven percent of subjects (63/862) were diagnosed with TMD pain. Both genders exhibited similar distributions of TMD diagnoses, except that myofascial pain was significantly more common in girls than in boys. Prevalence of pain once a week or more was reported as: 21% in the head; 12% in the temples; and 3% in the face, temporomandibular joint, or jaws. The prevalence of TMD-related pain was significantly higher in girls than in boys. Self-reported TMD symptoms were significantly more common (P < 0.001) in group 1. No significant gender differences were found in group 1 for pain intensity, behavioral rating scale scores, medicine consumption, reported days of school absence, or perceived need for treatment. Overall, TMD-related pain was more common in girls than in boys. A majority of children and adolescents who experienced pain once a week or more perceived a need for treatment. Seven percent of the examined subjects were diagnosed with TMD pain.


Saccharomyces boulardii Strain CNCM I-745 Modifies the Mononuclear Phagocytes Response in the Small Intestine of Mice Following Salmonella Typhimurium Infection.
Intestinal mononuclear phagocytes (MPs) comprise dendritic cells (DCs) and macrophages (Mφs) that play different roles in response to Salmonella infection. After phagocytosis, DCs expressing CD103 transport Salmonella from the intestinal tract to the mesenteric lymph nodes (MLN) and induce adaptive immune responses whereas resident Mφs expressing CX3CR1 capture bacteria in the lumen and reside in the lamina propria (LP) where they induce a local immune response. CX3CR1+ Mφs are generated from Ly6Chi monocytes that enter the colonic mucosa and differentiate locally. We previously demonstrated that the probiotic yeast Saccharomyces boulardii CNCM I-745 (S.b) prevents infection by Salmonella enterica serovar Typhimurium (ST), decreases ST translocation to the peripheral organs and modifies the pro-and anti-inflammatory cytokine profiles in the gut. In the present study, we investigated the effect of S.b on the migratory CD103+ DCs and the resident CX3CR1+ Mφs. MPs were isolated from the LP of streptomycin-treated mice infected by ST with or without S.b treatment before or during the infection. In S.b-pretreated mice, we observed a decrease of the CD103+ DCs in the LP that was associated with the drop of ST recovery from MLN. Interestingly, S.b induced an infiltration of LP by classical Ly6Chi monocytes, and S.b modified the monocyte-Mφ maturation process in ST-infected mice. Our results showed that S.b treatment induced the expansion of Ly6Chi monocytes in the blood as well as in the bone marrow (BM) of mice, thus contributing to the Mφ replenishment in LP from blood monocytes. In vitro experiments conducted on BM cells confirmed that S.b induced the expansion of CX3CR1+ Mφs and concomitantly ST phagocytosis. Altogether, these data demonstrate that Saccharomyces boulardii CNCM I-745 modulates the innate immune response. Although here, we cannot explicitly delineate direct effects on ST from innate immunity, S. b-amplified innate immunity correlated with partial protection from ST infection. This study shows that S.b can induce the expansion of classical monocytes that are precursors of resident Mφs in the LP.


Injury epidemiology in Iran: a systematic review.
Injuries are the second greatest cause of mortality in Iran. Information about the epidemiological pattern of injuries is effective in decision-making. In this regard, the aim of the current study is to elaborate on the epidemiology of injuries in Iran through a systematic review. Required data were collected searching the following key words and their Persian equivalents; trauma, injury, accident, epidemiology, prevalence, pattern, etiology, risk factors and Iran. The following databases were searched: Google Scholar, PubMed, Scopus, MagIran, Iranian scientific information database (SID) and Iran Medex. Some of the relevant journals and web sites were searched manually. The lists of references from the selected articles were also investigated. We have also searched the gray literature and consulted some experts. Out of 2747 retrieved articles, 25 articles were finally included in the review. A total of 3234481 cases have been investigated. Mean (SD) age among these cases was 30 (17.4) years. The males comprised 75.7% of all the patients. Only 31.1% of patients were transferred to hospital by ambulance. The most common mechanism of injuries was road traffic accidents (50.1%) followed by falls (22.3%). In road traffic accidents, motorcyclists have accounted for the majority of victims (45%). Roads were the most common accident scene for the injuries (57.5%). The most common injuries were to the head and neck. (47.3%). The mean (SD) Injury Severity Score (ISS) was 8.1(8.6%). The overall case-fatality proportion was 3.8% and 75% of all the mortalities related to road traffic accidents. The main priorities in reducing the burden of injuries include: the young, male target group, improving pre-hospital and ambulance services, preventing road traffic accidents, improving road safety and the safety of motorcyclists (compulsory helmet use, safer vehicles, dedicated motorcycle lanes).


Lithium: updated human knowledge using an evidence-based approach: part III: clinical safety.
Lithium use in mental diseases has changed over the years but remains a cornerstone of treatment in bipolar disorders. In two companion papers, we have reviewed existing (and especially recent) data on lithium efficacy and updated basic knowledge regarding the practical fundamentals of lithium therapy. The present paper reviews safety data on lithium available to date. Gastrointestinal pain or discomfort, diarrhoea, tremor, polyuria, nocturnal urination, weight gain, oedema, flattening of affect and exacerbation of psoriasis are typical complaints of patients receiving long-term lithium therapy. Renal involvement results in a reduced urinary concentrating capacity, expressed as obligate polyuria, with secondary thirst. With long-term therapy, this may result in nephrogenic diabetes insipidus. In addition, glomerular filtration rate falls slightly in about 20% of patients. The view that only a few patients receiving long-term lithium are at increased risk of glomerular impairment and progressive renal insufficiency should be regarded with caution. The risk is increased in case of concomitant diseases or medications. Lithium treatment may inhibit thyroid hormone release and induce goitre. Consequently, the prevalence of both overt and subclinical hypothyroidism is increased, with circulating thyroid auto-antibodies frequently being found. Much less commonly, thyrotoxicosis may also develop in association with lithium therapy. Long-term lithium treatment may also be associated with persistent hyperparathyroidism and hypercalcaemia, as well as with hypermagnesaemia. Overweight of up to 4-10 kg is found in approximately 30% of lithium-treated patients. Most neurological manifestations are benign, for example, the fine postural and/or action tremor present in 4-20% of patients. This is increased by high caffeine consumption and concomitant use of other psychotropic agents. A number of rare, potentially serious neurological adverse effects have been reported, including extrapyramidal symptoms, 'pseudotumour cerebri' or occasionally cerebellar symptoms. Severe neurological sequelae are exceptional. Cognitive disturbances are often mentioned as a lithium-related adverse effect. The few controlled studies do show a statistically significant negative effect of lithium on memory, vigilance, reaction time and tracking. There are frequent reports of mild effects of lithium on cognition at therapeutic serum concentrations. A number of deaths associated with lithium treatment have been reported. The most serious issue is that of non-accidental overdose, i.e. either long-term overdosage or acute overdose on long-term treatment. Progressive renal insufficiency, an exceptional complication of long-term lithium therapy, may also have a fatal outcome. In relation to pregnancy, lithium salts are rated as category D (positive evidence of risk). Therefore, prescription of lithium should be avoided during the first trimester of pregnancy unless the benefit to the mother exceeds the risk to the fetus. Although lithium transfer into breast milk is well established, the long-term fate of babies breast-fed by mothers receiving lithium therapy is unknown. Whether lithium therapy is safe in breast-feeding women is controversial. Although there is no absolute contraindication, it is known that the kidney is particularly sensitive to lithium just after birth. Intoxication in patients on long-term treatment with lithium in the absence of history of acute ingestion is not rare. Contributing factors include change in daily dose, long-term high dosage, kidney disease or drug interaction. In suspected cases, serum concentrations should be obtained early and repeatedly. In addition to supportive measures, haemodialysis is the treatment of choice for severe cases. Thorough knowledge of the limitations and drawbacks of lithium therapy is mandatory for its optimal use, especially at a time when its risk/benefit profile needs to be compared accurately with that of antiepileptic drugs and other mood stabilizing medications.


A nonparametric random coefficient approach for life expectancy growth using a hierarchical mixture likelihood model with application to regional data from North Rhine-Westphalia (Germany).
Life expectancy is of increasing prime interest for a variety of reasons. In many countries, life expectancy is growing linearly, without any indication of reaching a limit. The state of North Rhine-Westphalia (NRW) in Germany with its 54 districts is considered here where the above mentioned growth in life expectancy is occurring as well. However, there is also empirical evidence that life expectancy is not growing linearly at the same level for different regions. To explore this situation further a likelihood-based cluster analysis is suggested and performed. The modelling uses a nonparametric mixture approach for the latent random effect. Maximum likelihood estimates are determined by means of the EM algorithm and the number of components in the mixture model are found on the basis of the Bayesian Information Criterion. Regions are classified into the mixture components (clusters) using the maximum posterior allocation rule. For the data analyzed here, 7 components are found with a spatial concentration of lower life expectancy levels in a centre of NRW, formerly an enormous conglomerate of heavy industry, still the most densely populated area with Gelsenkirchen having the lowest level of life expectancy growth for both genders. The paper offers some explanations for this fact including demographic and socio-economic sources. This case study shows that life expectancy growth is widely linear, but it might occur on different levels.


Cloning of mouse RP-8 cDNA and its expression during apoptosis of lymphoid and myeloid cells.
A partial-length cDNA encoding a cell death-associated gene designated RP-8 was cloned from rat thymocytes by Owens and co-workers (Mol. Cell. Biol. 11, 4177-4188, 1991). They reported that transcription of RP-8 was induced when rat thymocytes were caused to undergo apoptosis triggered by either radiation or treatment with dexamethasone. To study the role of RP-8 in cell death in the mouse, we have cloned a full-length mouse RP-8 cDNA from a mouse thymus cDNA library. The mouse RP-8 gene encodes of protein of 343 amino acids with 92% amino acid identity to the rat RP-8 protein. Expression of mouse RP-8 was not altered in a factor-dependent myeloid line induced to undergo apoptosis by growth factor withdrawal, or when a lymphoid cell line was triggered to undergo apoptosis by irradiation. Although these data do not prove that RP-8 is not a cell death gene, they show that RP-8 expression is not sufficient for apoptosis, and that transcriptional up regulation of RP-8 is not universally associated with apoptosis.


Studies of the genetics, function, and kinetic mechanism of TagE, the wall teichoic acid glycosyltransferase in Bacillus subtilis 168.
The biosynthetic enzymes involved in wall teichoic acid biogenesis in gram-positive bacteria have been the subject of renewed investigation in recent years with the benefit of modern tools of biochemistry and genetics. Nevertheless, there have been only limited investigations into the enzymes that glycosylate wall teichoic acid. Decades-old experiments in the model gram-positive bacterium, Bacillus subtilis 168, using phage-resistant mutants implicated tagE (also called gtaA and rodD) as the gene coding for the wall teichoic acid glycosyltransferase. This study and others have provided only indirect evidence to support a role for TagE in wall teichoic acid glycosylation. In this work, we showed that deletion of tagE resulted in the loss of α-glucose at the C-2 position of glycerol in the poly(glycerol phosphate) polymer backbone. We also reported the first kinetic characterization of pure, recombinant wall teichoic acid glycosyltransferase using clean synthetic substrates. We investigated the substrate specificity of TagE using a wide variety of acceptor substrates and found that the enzyme had a strong kinetic preference for the transfer of glucose from UDP-glucose to glycerol phosphate in polymeric form. Further, we showed that the enzyme recognized its polymeric (and repetitive) substrate with a sequential kinetic mechanism. This work provides direct evidence that TagE is the wall teichoic acid glycosyltransferase in B. subtilis 168 and provides a strong basis for further studies of the mechanism of wall teichoic acid glycosylation, a largely uncharted aspect of wall teichoic acid biogenesis.


Motor activity in the stump of an amputated leg during free walking in cockroaches.
1. A rhythmic pattern of motor activity was recorded in the stump of an amputated leg during free walking in cockroaches. 2. During relatively rapid walking, extensor (depressor) and flexor (elevator) muscles in the intact coxa of the amputated leg showed one burst of activity during each cycle of bursting in an adjacent, intact leg. However, during slower walking these muscles could show two or three bursts of activity during each cycle of bursting in an intact leg. 3. Motor bursts in the stump of an amputated leg showed features similar to those of bursts recorded from intact legs. Burst duration increased with an increase in period, and the bursts generally showed consistent timing (phase) relative to bursts in most of the intact legs. 4. The motor pattern recorded in a stump was very like that recorded in an intact leg during walking, and unlike that recorded during searching for a foothold (defined in the text). It is concluded that after the amputation of most of a leg, motor neurones innervating muscles in the stump of the amputated leg continue to be driven by the interneurones that normally drive the intact leg during walking. Analysis of the motor pattern in the stump may therefore reveal important features of the locomotor control system.


Incomplete nonsense-mediated decay facilitates detection of a multi-exonic deletion mutation in SGCE.
Mutations in SGCE represent the major cause of the myoclonus-dystonia syndrome (DYT11), an autosomal dominant disorder of reduced penetrance. Virtually all affected individuals have myoclonus, which is concentrated in the upper extremities, neck and trunk. Over half of patients have dystonia, usually affecting the neck or arms. SGCE is maternally imprinted. Of the more than 70 SGCE mutations reported in the literature, 18 are large deletions disrupting at least one exon. Therefore, testing for exonic deletions should be considered in individuals with a classic phenotype in whom Sanger sequencing is unrevealing. However, standard methodologies for detection of exonic deletion mutations are expensive, labor intensive and can produce false negatives. Herein, we report the use of cDNA derived from leukocyte RNA to identify a deletion mutation (exons 4 and 5) of SGCE in a family with DYT11. Residual RNA from incomplete nonsense-mediated decay permitted reverse transcription to cDNA. Breakpoints of the 8939 bp heterozygous deletion were then defined with long-range polymerase chain reaction and Sanger sequencing. Use of cDNA generated by reverse transcription of leukocyte RNA can reduce the costs associated with diagnostic genetic testing and can facilitate detection of deletion mutations.


MiR-133a Mimic Alleviates T1DM-Induced Systolic Dysfunction in Akita: An MRI-Based Study.
Diabetic cardiomyopathy is a leading cause of heart failure. Developing a novel therapeutic strategy for diabetic cardiomyopathy and characterizing animal models used for diabetes mellitus (DM) are important. Insulin 2 mutant (Ins2+/-) Akita is a spontaneous, genetic, mouse model for T1DM, which is relevant to humans. There are contrasting reports on systolic dysfunction and pathological remodeling (hypertrophy and fibrosis) in Akita heart. Here, we used magnetic resonance imaging (MRI) approach, a gold standard reference for evaluating cardiac function, to measure ejection fraction (indicator of systolic dysfunction) in Akita. Moreover, we performed Wheat Germ Agglutinin (WGA) and hematoxylin and Eosin stainings to determine cardiac hypertrophy, and Masson's Trichrome and picrosirius red stainings to determine cardiac fibrosis in Akita. MiR-133a, an anti-hypertrophy and anti-fibrosis miRNA, is downregulated in Akita heart. We determined if miR-133a mimic treatment could mitigate systolic dysfunction and remodeling in Akita heart. Our MRI results revealed decreased ejection fraction in Akita as compared to WT and increased ejection fraction in miR-133a mimic-treated Akita. We also found that miR-133a mimic treatment mitigates T1DM-induced cardiac hypertrophy and fibrosis in Akita. We conclude that Akita shows cardiac hypertrophy, fibrosis and systolic dysfunction and miR-133a mimic treatment to Akita could ameliorate them.


The association between psoriasis, diabetes mellitus, and atherosclerosis in Israel: a case-control study.
Previous reports demonstrated an association between psoriasis and other diseases including heart failure and diabetes mellitus. Our aim was to describe the association between psoriasis, diabetes mellitus, and atherosclerosis in Israel. A cross-sectional study was performed utilizing the database of Maccabi Healthcare Services (MHS), a large health provider organization in Israel. Case patients were defined as subjects who were diagnosed with psoriasis. Patients with diabetes and atherosclerosis were identified by using the MHS diabetes and cardiovascular registries, respectively. The control group included MHS enrollees without psoriasis. The proportion of diabetes and atherosclerosis among case and control groups was compared. Chi-square tests were used to compare categorical parameters. Logistic regression models were used for multivariate analyses. The study included 46,095 patients with psoriasis (case patients) and 1,579,037 subjects without psoriasis (control patients). The age-adjusted proportion of diabetes was significantly higher in psoriasis patients as compared with the control group (odds ratio [OR] 1.27, 95% confidence interval [CI] 1.1-1.48). The age-adjusted proportion of atherosclerosis was significantly higher in psoriasis patients as compared with the control group (OR 1.28, 95% CI 1.04-1.59). In patients with psoriasis, a multivariate logistic regression model demonstrated an association between diabetes and the multiple use of very potent topical steroids (P < .05) or use of systemic medication for psoriasis (methotrexate, cyclosporine or acitretin) (P < .001). A similar model demonstrated an association between atherosclerosis and the use of phototherapy (P < .001). Our study was based on a computerized database. The diagnosis of psoriasis was based on digitally transmitted data. Therefore overestimation (false-positive cases) and underestimation (false-negative cases) of psoriasis patients may exist, thereby being a source for information bias. A second limitation is selection bias that may occur due to the possibility that reporting of both psoriasis and associated illnesses is higher in individuals who are seeking medical care. A third limitation concerns the causal effect between occurrence of psoriasis and atherosclerosis or diabetes. The dataset of MHS records diagnoses only from 1997 and does not record the date of disease onset. Our study supports previous reports for an association between psoriasis and atherosclerosis and psoriasis and diabetes. Further study is needed to support this observation.


Variations of lymphocyte sub-populations in vulvar condylomata during therapy with beta-interferon.
Several experiences induced us to consider genital HPV infection as an expression of a local immunodeficiency. The aim of our study was to research the effect of immunotherapy on the lymphocyte subpopulations and Langerhans cells in vulvar condyloma. Twenty women with persistent vulvar condylomata, treated with 2,000,000 IU/die of beta-interferon for 15 days, were submitted to vulvar biopsy before and 2-5 months after medical treatment. The frozen sections obtained were assayed with the following monoclonal antibodies: OKT4 (T helper lymphocytes), OKT8 (T suppressor lymphocytes), OKB7 (B lymphocytes) and S-100 protein (Langerhans cells). Using a morphometric evaluation, the average number of both intraepithelial and stromal lymphocyte subsets and of the intraepithelial Langerhans cells was assessed. In all the biopsies preceding the medical treatment we found a low number of T helper lymphocytes both in the epithelium and stroma, with inversion of T4/T8 lymphocyte ratio and rare presence of Langerhans cells. In patients with a good therapeutic response (50-100% of condyloma reduction) we observed an increase in intraepithelial T4 lymphocytes and a decrease in both intraepithelial and stromal T8 lymphocytes. In cases with persistent disease after therapy, the histological pattern was similar to that observed in the first biopsy, with the exception of a significant increase in the average number of Langerhans cells. Our data correlate the clinical response to the immunotherapy with the histology of lymphocyte subsets in the vulvar condylomata. The increase in Langerhans cells observed in patients with negative response may be interpreted with a probable inability of these cells to promote the immune reaction.


A Simple 3-Parameter Model for Examining Adaptation in Speech and Voice Production.
Sensorimotor adaptation experiments are commonly used to examine motor learning behavior and to uncover information about the underlying control mechanisms of many motor behaviors, including speech production. In the speech and voice domains, aspects of the acoustic signal are shifted/perturbed over time via auditory feedback manipulations. In response, speakers alter their production in the opposite direction of the shift so that their perceived production is closer to what they intended. This process relies on a combination of feedback and feedforward control mechanisms that are difficult to disentangle. The current study describes and tests a simple 3-parameter mathematical model that quantifies the relative contribution of feedback and feedforward control mechanisms to sensorimotor adaptation. The model is a simplified version of the DIVA model, an adaptive neural network model of speech motor control. The three fitting parameters of SimpleDIVA are associated with the three key subsystems involved in speech motor control, namely auditory feedback control, somatosensory feedback control, and feedforward control. The model is tested through computer simulations that identify optimal model fits to six existing sensorimotor adaptation datasets. We show its utility in (1) interpreting the results of adaptation experiments involving the first and second formant frequencies as well as fundamental frequency; (2) assessing the effects of masking noise in adaptation paradigms; (3) fitting more than one perturbation dimension simultaneously; (4) examining sensorimotor adaptation at different timepoints in the production signal; and (5) quantitatively predicting responses in one experiment using parameters derived from another experiment. The model simulations produce excellent fits to real data across different types of perturbations and experimental paradigms (mean correlation between data and model fits across all six studies = 0.95 ± 0.02). The model parameters provide a mechanistic explanation for the behavioral responses to the adaptation paradigm that are not readily available from the behavioral responses alone. Overall, SimpleDIVA offers new insights into speech and voice motor control and has the potential to inform future directions of speech rehabilitation research in disordered populations. Simulation software, including an easy-to-use graphical user interface, is publicly available to facilitate the use of the model in future studies.


Abundance and Distribution of Enteric Bacteria and Viruses in Coastal and Estuarine Sediments-a Review.
The long term survival of fecal indicator organisms (FIOs) and human pathogenic microorganisms in sediments is important from a water quality, human health and ecological perspective. Typically, both bacteria and viruses strongly associate with particulate matter present in freshwater, estuarine and marine environments. This association tends to be stronger in finer textured sediments and is strongly influenced by the type and quantity of clay minerals and organic matter present. Binding to particle surfaces promotes the persistence of bacteria in the environment by offering physical and chemical protection from biotic and abiotic stresses. How bacterial and viral viability and pathogenicity is influenced by surface attachment requires further study. Typically, long-term association with surfaces including sediments induces bacteria to enter a viable-but-non-culturable (VBNC) state. Inherent methodological challenges of quantifying VBNC bacteria may lead to the frequent under-reporting of their abundance in sediments. The implications of this in a quantitative risk assessment context remain unclear. Similarly, sediments can harbor significant amounts of enteric viruses, however, the factors regulating their persistence remains poorly understood. Quantification of viruses in sediment remains problematic due to our poor ability to recover intact viral particles from sediment surfaces (typically <10%), our inability to distinguish between infective and damaged (non-infective) viral particles, aggregation of viral particles, and inhibition during qPCR. This suggests that the true viral titre in sediments may be being vastly underestimated. In turn, this is limiting our ability to understand the fate and transport of viruses in sediments. Model systems (e.g., human cell culture) are also lacking for some key viruses, preventing our ability to evaluate the infectivity of viruses recovered from sediments (e.g., norovirus). The release of particle-bound bacteria and viruses into the water column during sediment resuspension also represents a risk to water quality. In conclusion, our poor process level understanding of viral/bacterial-sediment interactions combined with methodological challenges is limiting the accurate source apportionment and quantitative microbial risk assessment for pathogenic organisms associated with sediments in aquatic environments.


MDR1 synonymous polymorphisms alter transporter specificity and protein stability in a stable epithelial monolayer.
The drug efflux function of P-glycoprotein (P-gp) encoded by MDR1 can be influenced by genetic polymorphisms, including two synonymous changes in the coding region of MDR1. Here we report that the conformation of P-gp and its drug efflux activity can be altered by synonymous polymorphisms in stable epithelial monolayers expressing P-gp. Several cell lines with similar MDR1 DNA copy number were developed and termed LLC-MDR1-WT (expresses wild-type P-gp), LLC-MDR1-3H (expresses common haplotype P-gp), and LLC-MDR1-3HA (a mutant that carries a different valine codon in position 3435). These cell lines express similar levels of recombinant mRNA and protein. P-gp in each case is localized on the apical surface of polarized cells. However, the haplotype and its mutant P-gps fold differently from the wild-type, as determined by UIC2 antibody shift assays and limited proteolysis assays. Surface biotinylation experiments suggest that the non-wild-type P-gps have longer recycling times. Drug transport assays show that wild-type and haplotype P-gp respond differently to P-gp inhibitors that block efflux of rhodamine 123 or mitoxantrone. In addition, cytotoxicity assays show that the LLC-MDR1-3H cells are more resistant to mitoxantrone than the LLC-MDR1-WT cells after being treated with a P-gp inhibitor. Expression of polymorphic P-gp, however, does not affect the host cell's morphology, growth rate, or monolayer formation. Also, ATPase activity assays indicate that neither basal nor drug-stimulated ATPase activities are affected in the variant P-gps. Taken together, our findings indicate that "silent" polymorphisms significantly change P-gp function, which would be expected to affect interindividual drug disposition and response.


Examination of performance of the Center for Epidemiologic Studies Depression Scale Short Form 10 among African youth in poor, rural households.
Youth mental health has emerged as a pressing global issue. However, to advance research gaps in low-income settings, we need valid measures of common mental health disorders. Using primary data collected in five countries (Kenya, Malawi, Tanzania, Zambia, and Zimbabwe), this study aims to assess the psychometric properties of the commonly used 10-item Center for Epidemiological Studies Depression (CES-D 10) scale among poor, disadvantaged youth populations in sub-Saharan African (SSA). Youth samples from each country (sample sizes ranging from 651 to 2098) come from large household surveys with youth modules, collected for impact evaluations of cash transfer programs targeted to poor families. For each sample, we assessed internal consistency (alpha), conducted factor analysis, and then examined construct validity and measurement invariance. We performed both exploratory (EFA) and confirmatory factor analysis (CFA) to examine and confirm the structure of the CES-D 10 for each country and then used multigroup CFA to assess measurement invariance across gender and age. Multivariate analyses were conducted to assess construct validity via test of the relationship between CES-D 10 and background characteristics. Results show the CES-D 10 had strong psychometric properties and was a reliable measure of depressive symptoms among disadvantaged youth in SSA. Across countries, there was high internal consistency (Cronbach alphas = 0.70-0.76) and the traditional two-factor solution showed good model fit. Full measurement invariance of the CES-D 10 was supported across gender. Consistent with previous literature on risk factors for depressive symptoms, the CES-D 10 was associated with increasing age, and female gender and being out of school in some locations. Results from this study support broad use of the CES-D 10 among poor youth populations in SSA. Between one-third and two-thirds of our samples demonstrated depressive symptoms as classified by recommended cut-offs for the CES-D 10, indicating a high burden of mental illness in disadvantaged youth populations. This tool can be used in future efforts to study prevalence and dynamics of depressive symptoms in this population, as well as effectiveness of policies and interventions to improve the mental health of youth in SSA.


Internet addiction disorder and problematic use of Google Glass™ in patient treated at a residential substance abuse treatment program.
Internet addiction disorder (IAD) is characterized by the problematic use of online video games, computer use, and mobile handheld devices. While not officially a clinical diagnosis according to the most recent version of the Diagnostic and Statistical Manual of Mental Disorders (DSM), individuals with IAD manifest severe emotional, social, and mental dysfunction in multiple areas of daily activities due to their problematic use of technology and the internet. We report a 31year-old man who exhibited problematic use of Google Glass™. The patient has a history of a mood disorder most consistent with a substance induced hypomania overlaying a depressive disorder, anxiety disorder with characteristics of social phobia and obsessive compulsive disorder, and severe alcohol and tobacco use disorders. During his residential treatment program at the Navy's Substance Abuse and Recovery Program (SARP) for alcohol use disorder, it was noted that the patient exhibited significant frustration and irritability related to not being able to use his Google Glass™. The patient exhibited a notable, nearly involuntary movement of the right hand up to his temple area and tapping it with his forefinger. He reported that if he had been prevented from wearing the device while at work, he would become extremely irritable and argumentative. Over the course of his 35-day residential treatment, the patient noted a reduction in irritability, reduction in motor movements to his temple to turn on the device, and improvements in his short-term memory and clarity of thought processes. He continued to intermittently experience dreams as if looking through the device. To our knowledge, this is the first reported case of IAD involving problematic use of Google Glass™.


Mechanical Genomic Studies Reveal the Role of d-Alanine Metabolism in Pseudomonas aeruginosa Cell Stiffness.
The stiffness of bacteria prevents cells from bursting due to the large osmotic pressure across the cell wall. Many successful antibiotic chemotherapies target elements that alter mechanical properties of bacteria, and yet a global view of the biochemistry underlying the regulation of bacterial cell stiffness is still emerging. This connection is particularly interesting in opportunistic human pathogens such as Pseudomonas aeruginosa that have a large (80%) proportion of genes of unknown function and low susceptibility to different families of antibiotics, including beta-lactams, aminoglycosides, and quinolones. We used a high-throughput technique to study a library of 5,790 loss-of-function mutants covering ~80% of the nonessential genes and correlated P. aeruginosa individual genes with cell stiffness. We identified 42 genes coding for proteins with diverse functions that, when deleted individually, decreased cell stiffness by >20%. This approach enabled us to construct a "mechanical genome" for P. aeruginosa d-Alanine dehydrogenase (DadA) is an enzyme that converts d-Ala to pyruvate that was included among the hits; when DadA was deleted, cell stiffness decreased by 18% (using multiple assays to measure mechanics). An increase in the concentration of d-Ala in cells downregulated the expression of genes in peptidoglycan (PG) biosynthesis, including the peptidoglycan-cross-linking transpeptidase genes ponA and dacC Consistent with this observation, ultraperformance liquid chromatography-mass spectrometry analysis of murein from P. aeruginosa cells revealed that dadA deletion mutants contained PG with reduced cross-linking and altered composition compared to wild-type cells.IMPORTANCE The mechanical properties of bacteria are important for protecting cells against physical stress. The cell wall is the best-characterized cellular element contributing to bacterial cell mechanics; however, the biochemistry underlying its regulation and assembly is still not completely understood. Using a unique high-throughput biophysical assay, we identified genes coding proteins that modulate cell stiffness in the opportunistic human pathogen Pseudomonas aeruginosa This approach enabled us to discover proteins with roles in a diverse range of biochemical pathways that influence the stiffness of P. aeruginosa cells. We demonstrate that d-Ala-a component of the peptidoglycan-is tightly regulated in cells and that its accumulation reduces expression of machinery that cross-links this material and decreases cell stiffness. This research demonstrates that there is much to learn about mechanical regulation in bacteria, and these studies revealed new nonessential P. aeruginosa targets that may enhance antibacterial chemotherapies or lead to new approaches.


Robust Fitting of a Weibull Model with Optional Censoring.
The Weibull family is widely used to model failure data, or lifetime data, although the classical two-parameter Weibull distribution is limited to positive data and monotone failure rate. The parameters of the Weibull model are commonly obtained by maximum likelihood estimation; however, it is well-known that this estimator is not robust when dealing with contaminated data. A new robust procedure is introduced to fit a Weibull model by using L2 distance, i.e. integrated square distance, of the Weibull probability density function. The Weibull model is augmented with a weight parameter to robustly deal with contaminated data. Results comparing a maximum likelihood estimator with an L2 estimator are given in this article, based on both simulated and real data sets. It is shown that this new L2 parametric estimation method is more robust and does a better job than maximum likelihood in the newly proposed Weibull model when data are contaminated. The same preference for L2 distance criterion and the new Weibull model also happens for right-censored data with contamination.


Pain prevalence and risk distribution among inpatients in a German teaching hospital.
This study was designed to provide a cross-sectional analysis of pain prevalence, chronicity, and severity as well as the impact of pain on psychological and social variables in inpatients in various departments of a German teaching hospital. Patients were asked to complete a questionnaire including sections on sociodemographic and socioeconomic data, pain variables, recent and past health care utilization, and screening questionnaires for depression, anxiety, and health-related quality of life. Of the 438 patients, 386 (88.1%) had experienced pain in the past 12 months; 367 (83.8%) reported having pain in the previous 3 months. Sixty-four percent of the pain patients stated that pain was the main reason for hospital admission; 48% reported having three or more pain sites. The most common location of pain was the back (26.9%). Pain patients showed significantly higher depression and anxiety scores and markedly reduced physical health when compared to non-pain patients. The results of this study indicate that in most medical disciplines pain is more than merely a symptom of disease. In many instances pain should be considered a serious comorbidity that can influence the outcome of medical and surgical treatment. Recent research has shown that prevention of the pain chronification process is the most promising strategy for avoiding the development of intractable pain. Acceptance, recognition, and assessment of pain as a risk factor at an early stage are essential factors. A first step might involve routine screening for pain on admission to any hospital facility, and subsequently evaluating the impact of pain on biopsychosocial functions.


Myosin subfragment-1 attachment to actin. Expected effect on equatorial reflections.
The characteristic equatorial X-ray pattern from a relaxed vertebrate skeletal muscle changes when the muscle is activated. In particular, there is a simultaneous decrease in the intensity of the first reflection (I10) and increase in the intensity of the second (I11). This observed change is almost reciprocal. When compared with the predictions of computer modeling, it produces a strong argument that the intensity change is due to a redistribution of myosin heads (myosin subfragment-1 or S-1), which results from the formation and configuration changes of actin-myosin links. Computer modeling shows that different actin-S-1 configurations will give different numerical values for I10 and I11, assuming the same number of attachments. For a given configuration, the intensity changes are a nonlinear function of attachment number, so that direct scaling of force to reflection intensity may be difficult. Data from active muscle are consistent with the notion that in different states of active muscle, i.e. shortening or isometric, there are different average configurations of actin-myosin attachment and different numbers of actin-myosin links.


Intra-residue interactions in proteins: interplay between serine or cysteine side chains and backbone conformations, revealed by laser spectroscopy of isolated model peptides.
Intra-residue interactions play an important role in proteins by influencing local folding of the backbone. Taking advantage of the capability of gas phase experiments to provide relevant information on the intrinsic H-bonding pattern of isolated peptide chains, the intra-residue interactions of serine and cysteine residues, i.e., OH/SH···OC(i) C6 and NH(i···)O/S C5 interactions in Ser/Cys residues, are probed by laser spectroscopy of isolated peptides. The strength of these local side chain-main chain interactions, elegantly documented from their IR spectral features for well-defined conformations of the main chain, demonstrates that a subtle competition exists between the two types of intra-residue bond: the C6 H-bond is the major interaction with Ser, in contrast to Cys where C5 interaction takes over. The restricted number of conformers observed in the gas phase experiment with Ser compared to Cys (where both extended and folded forms are observed) also suggests a significant mediation role of these intra-residue interactions on the competition between the several main chain folding patterns.


United States Military Parachute Injuries: Part 2: Interventions Reducing Military Parachute Injuries in Training and Operations.
This is part 2 of an article detailing the reduction in airbornerelated injuries over time. Part 1 examined the early history of airborne operations and provided evidence for the reduction in injuries over time; part 2 discusses interventions associated with the decline in injury rates. In 1943 at the United States (US) Army Airborne School, data showed that injuries were substantially reduced from 120 to 18 injuries/1000 trainees. Credit for the reduction was given to development of the parachute landing fall (PLF), better supervision of students while in initial airborne training, intensive ground training prior to actual jumping, and elimination of dangerous and unnecessary training procedures (like practice jumps from 11-foot heights). Compared to the older T-10 parachute introduced in the 1950s, the newer T-11 parachute introduced in 2010 reduced injuries by 43% in operational training (9.1 vs 5.2 injuries/1000 jumps). In aircraft with jump doors on both sides, alternating jumps between the doors so that the jumpers exit at slightly different times reduced high-altitude and mid-altitude entanglement injuries by 85% (0.13 to 0.02 injury/ 1000 jumps). Data from six scientific studies involving more than 1,300,000 jumps and two systematic reviews indicated that the parachute ankle brace (PAB) reduced ankle injuries and ankle fractures by about half with an estimated return on investment of at least $7 in medical and personnel costs for every $1 spent on the PAB. However, the PAB is not currently used or even well-known within the airborne community because of a lack of acceptance and promotion. While some airborne injury-reducing innovations are discussed here it is likely that there have been others that have not been documented. It is important to detail these interventions so future paratroopers and leaders can better understanding their rationale and effectiveness.


Oxytocin improves cytological and histological profiles of vaginal atrophy in postmenopausal women.
To investigate if topical oxytocin can reverse vaginal atrophy, as assessed by cytological and histological examination of the vaginal mucosal epithelium, in postmenopausal women after 12 weeks of treatment as compared to placebo. Sixty-eight postmenopausal women diagnosed with vaginal atrophy were randomized for this multicenter, double-blinded, placebo-controlled trial. Thirty-three women received 600 IU vagitocin, an oxytocin containing gel, and 35 women received a placebo gel intravaginally. The dose was 600 IU daily for the first two weeks and thereafter 600 IU twice a week for 10 weeks. All participant women underwent four visits and a subgroup of 20 women had a further fifth visit. Vaginal smears for cytological evaluation were collected at all visits. Vaginal biopsies were taken in 20 women before and after 12 weeks of treatment for histological analysis. In these women a vaginal smear was also collected after 14 weeks. The increase in the percentage of superficial cells between 0 and 2 weeks was significantly greater after treatment with vagitocin in comparison with placebo (p = 0.04). The difference in the maturation value between 0 and 12 weeks was significantly higher in the vagitocin than in the placebo group (p = 0.01). The reduction in the scores of atrophy was according to the histological investigation significantly greater in the vagitocin group than in the placebo group at 12 weeks (p < 0.04). Daily intravaginal treatment with vagitocin 600 IU improves expressions of vaginal atrophy as recorded by cytological investigation of vaginal smears and histological analysis of vaginal biopsies. Treatment twice weekly seems to be less effective regarding the increase in superficial cells.


Do Model-Based Studies in Chronic Obstructive Pulmonary Disease Measure Correct Values of Utility? A Meta-Analysis.
Chronic obstructive pulmonary disease (COPD) is a progressive chronic disease that has considerable impact on utility-based health-related quality of life. Utility is a key input of many decision analytic models used for economic evaluations. To systematically review COPD-related utilities and to compare these with alternative values used in decision models. The literature review comprised studies that generated utilities for COPD-related stages based on EuroQol five-dimensional questionnaire surveys of patients and of decision models of COPD progression that have been used for economic evaluations. The utility values used in modeling studies and those from the meta-analysis of actual patient-level studies were compared and differences quantified. Twenty decision modeling studies that used utility value as an input parameter were found. Within the same span of publication period, 13 studies involving patient-level utility data were identified and included in the meta-analysis. The estimated mean utility values ranged from 0.806 (95% confidence interval [CI] 0.747-0.866) for stage I to 0.616 (95% CI 0.556-0.676) for stage IV. The utility scores for comparable stages in modeling studies were different (significant difference 0.045 [95% CI 0.041-0.052] for stage III). Modeling studies consistently used higher utility values than the average reported patient-level data. COPD decision analytic models are based on a limited range of utility values that are systematically different from average values estimated using a meta-analysis. A more systematic approach in the application of utility measures in economic evaluation is required to appropriately reflect current literature.


Extended major histocompatibility complex haplotypes in type I diabetes mellitus.
We have studied major histocompatibility complex markers in Caucasian patients with type I diabetes mellitus and their families. The frequencies of extended haplotypes that were composed of specific HLA-B, HLA-DR, BF, C2, C4A, and C4B allelic combinations, which occurred more commonly than expected, were compared on random diabetic and normal chromosomes in the study families. We demonstrated that all of the previously recognized increases in HLA-B8, B18, B15, DR3, and perhaps DR4 could be ascribed to the increase among diabetic haplotypes of a few extended haplotypes: [HLA B8, DR3, SC01, GLO2]; [HLA-B18, DR3, F1C30]; [HLA-B15, DR4, SC33]; and [HLA-BW38, DR4, SC21]. In fact, HLA-DR3 on nonextended haplotypes was "protective", with a relative risk considerably less than 1.0. There was a paucity or absence among diabetic patients of several extended haplotypes of normal chromosomes, notably [HLA-B7, DR2, SC31] and [HLA-BW44, DR4, SC30]. The extended haplotype [HLA-BW38, DR4, SC21] is found only in Ashkenazi Jewish patients, which suggests that extended haplotypes mark specific mutations that arise in defined ethnic groups. The data show that no known MHC allele, including HLA-DR3 and possibly HLA-DR4, is per se a marker for or itself a susceptibility gene for type I diabetes. Rather, extended haplotypes, with relatively fixed alleles, are either carriers or noncarriers of susceptibility genes for this disease. Thus, the increased frequency (association) or the decreased frequency (protection) of individual MHC alleles is largely explainable by these extended haplotypes.


Multimodal imaging demonstrates concomitant changes in bone and cartilage after destabilisation of the medial meniscus and increased joint laxity.
Alterations in joint mechanics can cause osteoarthritis, which results in degeneration of both cartilage and bone tissue. The objective of this work is to measure changes in the laxity of the mouse knee joint after destabilisation of the medial meniscus (DMM) and to visualise and quantify the resulting three-dimensional changes in the bone and cartilage. Skeletally mature C57Bl6 male mice underwent DMM surgery in the right leg. Animals were sacrificed immediately 0 weeks (n=15), 4 weeks (n=11) or 8 weeks (n=12) after surgery. For the 0-week group, the anterior-posterior (AP) and varus-valgus laxity of the DMM limb were compared to the contralateral limb. For 4 and 8-week groups, tibiae were scanned with micro-computed tomography (μCT) to quantify and visualise bone changes and with confocal scanning laser microscopy (CSLM) to measure changes in cartilage. Laxity testing measured an increase in AP range of motion, particularly in the anterior direction. The DMM limbs showed a decrease in epiphyseal trabecular bone at 8 weeks and a decrease in cartilage volume, primarily on the posterior medial plateau, compared to the contralateral limb. Significant bone remodelling was observed at the periphery of the joint and in severe cases, osteolysis extended through the growth plate. Multimodal imaging allowed quantifiable 3D assessment of bone and cartilage and indicated extensive changes in the tissues. The increase in AP laxity suggests that DMM surgery redistributes loading posteriorly on the medial plateau, resulting in bone and cartilage loss primarily on the posterior portion of the medial plateau.


Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells.
Ca(2+) and calmodulin (CaM) play a critical role in proliferation and viability of a wide variety of cells, including prostate cancer cells. We examined two prostate cancer cell lines, androgen-sensitive LNCaP and androgen-independent PC-3. Proliferation of LNCaP cells was six to eight times more sensitive to the inhibitory effect of the CaM antagonist N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7) than were PC-3 cells. Because LNCaP cell proliferation is sensitive to stimulation by androgen, we assessed the physical and functional interaction between androgen receptor (AR) and CaM. We observed tight binding of AR to CaM when LNCaP cell extracts were subjected to CaM-affinity column chromatography. AR binding to CaM was Ca(2+)-dependent and was inhibited by pretreatment of the cell extracts with W-7. Using immunofluorescence staining and confocal microscopy, we demonstrated colocalization of AR and CaM in the nucleus of LNCaP cells. Furthermore, the functional relevance of AR-CaM interactions in intact cells was revealed by the observation that W-7 was as effective as Casodex, an antiandrogen, in blocking AR-regulated expression of prostate-specific antigen in LNCaP cells. AR seems to interact with CaM directly because purified human AR could bind to CaM-agarose, and CaM could be detected in AR-immunoprecipitate prepared from purified soluble proteins. These studies provide direct evidence for physical and functional interaction between AR and CaM and suggest the potential usefulness of CaM antagonists in blocking AR activity in prostate cancer.


Human local adaptation of the TRPM8 cold receptor along a latitudinal cline.
Ambient temperature is a critical environmental factor for all living organisms. It was likely an important selective force as modern humans recently colonized temperate and cold Eurasian environments. Nevertheless, as of yet we have limited evidence of local adaptation to ambient temperature in populations from those environments. To shed light on this question, we exploit the fact that humans are a cosmopolitan species that inhabit territories under a wide range of temperatures. Focusing on cold perception-which is central to thermoregulation and survival in cold environments-we show evidence of recent local adaptation on TRPM8. This gene encodes for a cation channel that is, to date, the only temperature receptor known to mediate an endogenous response to moderate cold. The upstream variant rs10166942 shows extreme population differentiation, with frequencies that range from 5% in Nigeria to 88% in Finland (placing this SNP in the 0.02% tail of the FST empirical distribution). When all populations are jointly analyzed, allele frequencies correlate with latitude and temperature beyond what can be explained by shared ancestry and population substructure. Using a Bayesian approach, we infer that the allele originated and evolved neutrally in Africa, while positive selection raised its frequency to different degrees in Eurasian populations, resulting in allele frequencies that follow a latitudinal cline. We infer strong positive selection, in agreement with ancient DNA showing high frequency of the allele in Europe 3,000 to 8,000 years ago. rs10166942 is important phenotypically because its ancestral allele is protective of migraine. This debilitating disorder varies in prevalence across human populations, with highest prevalence in individuals of European descent-precisely the population with the highest frequency of rs10166942 derived allele. We thus hypothesize that local adaptation on previously neutral standing variation may have contributed to the genetic differences that exist in the prevalence of migraine among human populations today.


Using High-Fidelity Patient Simulation and an Advanced Distance Education Network to Teach Pharmacology to Second-Year Medical Students.
The feasibility and acceptance of an Advanced Distance Education Network (ADEN) in bringing the simulated operating room (OR) to second-year medical students learning the pharmacology of anesthetic drugs is reviewed. A MedSim-Eagle (Binghamton, NY) full-scale mannequin simulator was used. Using an ADEN, students were linked in real time to a simulated OR where the anesthesiologist instructor was using a MedSim-Eagle patient simulator to present for discussion the physiologic effects of volatile anesthetics on cardiac output (CO), heart rate (HR), mean arterial pressure (MAP), and systemic vascular resistance (SVR). The use of simulation to present basic science principles of isoflurane and halothane's effect on CO, HR, MAP, and SVR in a clinical setting via an ADEN is feasible. Student acceptance of this method of education is high, as measured by a post-exercise survey. Ninety-five percent of students felt this exercise was a valuable use of their time; 93% felt the ADEN-delivered clinical simulation presentation contributed to their understanding of the pharmacology of anesthesia. Eighty-three percent of students preferred this integrated clinically oriented review to a didactic review of the material, and 92% of students who had experienced previous small group hands-on session simulation felt the ADEN-delivered session was the same or better.


Inorganic-organic hybrid scaffolds for osteochondral regeneration.
Ligament graft failure frequently results from poor integration of the replacement tissue with associated bone. Thus, the ability to regenerate the bone-ligament osteochondral interface would be advantageous in ligament reconstruction. At the osteochondral interface, the tissue transitions from a bone-like matrix to fibrocartilage. Therefore, a scaffold which promotes a spatially regulated transition in cell behavior from osteoblast-like to chondrocyte-like would be desirable. Previous research indicates that addition of inorganic components to organic scaffolds can enhance the deposition of bone-like matrix by associated osteoblasts. We therefore reasoned that a gradient in the inorganic content of a hybrid inorganic-organic scaffold may induce an osteochondral-like transition in cell phenotype and matrix production. To test this hypothesis, hydrogels were prepared from poly(ethylene glycol) (PEG) and star poly(dimethylsiloxane) (PDMS(star)). As anticipated, both the matrix deposition and phenotype of encapsulated osteoblasts varied with scaffold inorganic content, although the directionality of this modulation was contrary to expectation. Specifically, osteoblasts appeared to transdifferentiate into chondrocyte-like cells with increasing scaffold inorganic content, as indicated by increased chondroitin sulfate and collagen type II production and by upregulation of sox9, a transcription factor associated with chondrocytic differentiation. Furthermore, the deposition of bone-like matrix (collagen type I, calcium phosphate, and osteocalcin) decreased with increasing PDMS(star) content. The resistance of the PDMS(star)-PEG scaffolds to protein adsorption and/or the changes in gel modulus/mesh structure accompanying PDMS(star) incorporation may underlie the unexpected increase in chondrocytic phenotype with increasing inorganic content. Combined, the present results indicate that PDMS(star)-PEG hybrid gels may prove promising for osteochondral regeneration. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.


Unidirectional supramolecular self-assembly inside nanocorrals via in situ STM nanoshaving.
Self-assembly of an alkylated diacetylene derivative is spatially confined via in situ scanning tunneling microscopy (STM) nanoshaving inside covalently modified highly ordered pyrolytic graphite (CM-HOPG). In contrast to unconstrained self-assembly that occurs randomly along three thermodynamically equivalent surface lattice directions, spatially confined assemblies are shown to form along chosen substrate orientations. Experimental statistics suggest two mechanisms for breaking the rotational degeneracy of the surface. First, the assembly orientation is biased via lateral confinement inside nanocorrals that do not match the substrate symmetry. Second, an interaction between the assembling molecules and the STM tip during nanoshaving guides 2D crystal nucleation and growth. The results presented here open new possibilities to regulate and orient self-assembled architectures via in situ nanomechanical manipulation techniques and provide mechanistic insights into the process.


Selective inhibition of BRCA2-deficient mammary tumor cell growth by AZD2281 and cisplatin.
To assess efficacy of the novel, selective poly(ADP-ribose) polymerase-1 (PARP-1) inhibitor AZD2281 against newly established BRCA2-deficient mouse mammary tumor cell lines and to determine potential synergy between AZD2281 and cisplatin. We established and thoroughly characterized a panel of clonal cell lines from independent BRCA2-deficient mouse mammary tumors and BRCA2-proficient control tumors. Subsequently, we assessed sensitivity of these lines to conventional cytotoxic drugs and the novel PARP inhibitor AZD2281. Finally, in vitro combination studies were done to investigate interaction between AZD2281 and cisplatin. Genetic, transcriptional, and functional analyses confirmed the successful isolation of BRCA2-deficient and BRCA2-proficient mouse mammary tumor cell lines. Treatment of these cell lines with 11 different anticancer drugs or with gamma-irradiation showed that AZD2281, a novel and specific PARP inhibitor, caused the strongest differential growth inhibition of BRCA2-deficient versus BRCA2-proficient mammary tumor cells. Finally, drug combination studies showed synergistic cytotoxicity of AZD2281 and cisplatin against BRCA2-deficient cells but not against BRCA2-proficient control cells. We have successfully established the first set of BRCA2-deficient mammary tumor cell lines, which form an important addition to the existing preclinical models for BRCA-mutated breast cancer. The exquisite sensitivity of these cells to the PARP inhibitor AZD2281, alone or in combination with cisplatin, provides strong support for AZD2281 as a novel targeted therapeutic against BRCA-deficient cancers.


O-GlcNAcylation of TDP-43 suppresses proteinopathies and promotes TDP-43's mRNA splicing activity.
Pathological TDP-43 aggregation is characteristic of several neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-TDP); however, how TDP-43 aggregation and function are regulated remain poorly understood. Here, we show that O-GlcNAc transferase OGT-mediated O-GlcNAcylation of TDP-43 suppresses ALS-associated proteinopathies and promotes TDP-43's splicing function. Biochemical and cell-based assays indicate that OGT's catalytic activity suppresses TDP-43 aggregation and hyperphosphorylation, whereas abolishment of TDP-43 O-GlcNAcylation impairs its RNA splicing activity. We further show that TDP-43 mutations in the O-GlcNAcylation sites improve locomotion defects of larvae and adult flies and extend adult life spans, following TDP-43 overexpression in Drosophila motor neurons. We finally demonstrate that O-GlcNAcylation of TDP-43 promotes proper splicing of many mRNAs, including STMN2, which is required for normal axonal outgrowth and regeneration. Our findings suggest that O-GlcNAcylation might be a target for the treatment of TDP-43-linked pathogenesis.


The endocannabinoid system: an emerging key player in inflammation.
The purpose of this review is to illustrate the expanding view of the endocannabinoid system (ECS) in relation to its roles in inflammation. According to the formal classification, the ECS consists of two cannabinoid receptors, their endogenous fatty acid-derived ligands, and a number of enzymes involved in their synthesis and breakdown. However, many endogenous congeners of classical endocannabinoids have now been discovered, together with a set of receptors structurally or functionally related to the cannabinoid receptors. Endocannabinoids per se behave 'promiscuously' with regard to their receptor interactions. It is increasingly recognized how tightly this expanded ECS is intertwined with key processes involved in inflammation. A continuous dynamic exchange of substrates and metabolites exists between ECS and eicosanoid pathways. Endocannabinoids can also be oxygenated by cyclooxygenase and other enzymes to biologically active 'hybrid' structures. Diet is among the main factors determining synthesis and release of endocannabinoids and related mediators. The complexity of what may be called the 'endocannabinoidome' requires approaches that take into account its dynamics and interconnections with other regulatory systems. This endocannabinoidome continues to offer possibilities for prevention and intervention, but multiple target approaches will probably provide the only keys to success.


Males and females gain differentially from sociality in a promiscuous fruit bat Cynopterus sphinx.
Sociality emerges when the benefits of group living outweigh its costs. While both males and females are capable of strong social ties, the evolutionary drivers for sociality and the benefits accrued maybe different for each sex. In this study, we investigate the differential reproductive success benefits of group membership that males and females might obtain in the promiscuous fruit bat Cynopterus sphinx. Individuals of this species live in flexible social groups called colonies. These colonies are labile and there is high turnover of individuals. However, colony males sire more offspring within the colony suggesting that being part of a colony may result in reproductive benefits for males. This also raises the possibility that long-term loyalty towards the colony may confer additional advantage in terms of higher reproductive success. We used ten seasons of genetic parentage data to estimate reproductive success and relatedness of individuals in the colony. We used recapture data to identify long and short-term residents in the colony as well as to obtain rates of recapture for males and females. Our results reveal that males have a significantly higher chance of becoming long-term residents (than females), and these long-term resident males gain twice the reproductive success compared to short-term resident males. We also observed that long-term resident females are related to each other and also achieve higher reproductive success than short-term resident females. In contrast, long-term resident males do not differ from short-term resident males in their levels of relatedness. Our results re-iterate the benefits of sociality even in species that are promiscuous and socially labile and possible benefits of maintaining a colony.


A randomized, controlled, dose-guiding study of the oral direct thrombin inhibitor ximelagatran compared with standard therapy for the treatment of acute deep vein thrombosis: THRIVE I.
This randomized, controlled, multicentre study evaluated the efficacy and tolerability of the oral direct thrombin inhibitor ximelagatran, compared with a low-molecular-weight heparin (dalteparin) followed by warfarin, in the treatment of deep vein thrombosis (DVT) of the lower extremity. Patients with acute DVT received oral ximelagatran (24, 36, 48 or 60 mg twice daily) or dalteparin and warfarin for 2 weeks. Evaluation of paired venograms from 295 of 350 patients showed regression of the thrombus in 69% of patients treated with ximelagatran and 69% of patients treated with dalteparin and warfarin. Progression was observed in 8% and 3% of patients, respectively. Changes in thrombus size according to the Marder score were similar in all groups. Treatment discontinuation due to bleeding occurred in two patients receiving ximelagatran (24- and 36-mg groups) and in two patients receiving dalteparin and warfarin. Reduction in pain, edema and circumference of the affected leg was similar in all groups. Oral ximelagatran appears to be a promising alternative to current anticoagulant therapy to limit the progression of acute DVT, and it seems to possess a wide therapeutic window.


Experiences of and actions towards worries among ambulance nurses in their professional life: a critical incident study.
Nurses working in the ambulance service are frequently exposed to situations in which they have to take sole responsibility for sick or injured patients, many of which can be emotionally trying. The purpose of this study was therefore to describe critical incidents in which ambulance nurses experience worry in their professional life and the actions they take in order to prevent and cope with it. The sample consisted of 13 male and 12 female nurses working in ambulance services in three small to medium-sized counties in the south of Sweden. The data were collected by means of interviews and analysed according to Critical Incident Technique (CIT). The result showed that nurses worried about specific emergency situations as well as situations related to their working environment. Worry was alleviated by their own actions or with help from others. It was also shown that, with increasing experience, came a responsibility to be able to cope with all kinds of situations. This responsibility was experienced as worrying. However, if the nurse felt confident in their colleague, the worry could ease. It was concluded that the worry experienced by ambulance nurses in specific emergency situations requires flexibility on the part of the ambulance service. Nurses must be given the opportunity to address their worries in the way that is most appropriate for each individual.