Polymorphism at the -308-promoter position of the tumor necrosis factor-alpha (TNF-alpha) gene and cervical cancer.
The purpose of the study was to investigate the hypothesis that the genetically programmed ability to produce low, medium, or high levels of tumor necrosis factor-alpha (TNF-alpha), as determined by TNF-alpha promoter polymorphism at position 308, influenced the development of cancer of the uterine cervix. The population was recruited from patients attending gynecological clinics at two teaching hospitals in Harare, Zimbabwe. Laboratory tests were performed in the Departments of Immunology and Medical Microbiology, Medical School, University of Zimbabwe. One hundred and three patients with invasive cancer of the uterine cervix and 101 healthy women were included in the study. All patients and healthy controls were from the Shona ethnic groups that inhabit northern Zimbabwe. DNA was purified from cervical cytobrush samples obtained from women with cervical cancer. In random cases a second DNA sample was extracted from patient blood. Control DNA was extracted from urine or peripheral blood samples from the healthy women. Detection of allele A and /or G at the 308 position in the promoter region of the TNF-alpha gene was carried out using the amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) technique. Polymorphism in the amplified products was detected by gel electrophoresis. There was no statistically significant difference in the distribution of the low (G) or high (A) producer alleles at position 308 of the TNF-alpha gene between patients with cervical cancer and healthy women. The high producer haplotype AA was identified in only one patient with cervical cancer and two healthy women. These data suggest that the genetically acquired ability to produce higher levels of TNF-alpha is present in a minority of women with or without cervical cancer in the Zimbabwean population. Homozygosity for allele 308A is very rare. High-producer allele 308A as well as high-producer haplotypes AA is significantly less common in a Zimbabwean population than in a European population.


Longitudinal effects of perinatal social support on maternal depression: a marginal structural modelling approach.
Depression in the perinatal period, during pregnancy or within 1 year of childbirth, imposes a high burden on women with rippling effects through her and her child's life course. Social support may be an important protective factor, but the complex bidirectional relationship with depression, alongside a paucity of longitudinal explorations, leaves much unknown about critical windows of social support exposure across the perinatal period and causal impacts on future depressive episodes. This study leverages marginal structural models to evaluate associations between longitudinal patterns of perinatal social support and subsequent maternal depression at 6 and 12 months postpartum. In a cohort of women in rural Pakistan (n=780), recruited in the third trimester of pregnancy and followed up at 3, 6 and 12 months postpartum, we assessed social support using two well-validated measures: the Multidimensional Scale of Perceived Social Support (MSPSS) and the Maternal Social Support Index (MSSI). Major depressive disorder was assessed with the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders (DSM IV). High and sustained scores on the MSPSS through the perinatal period were associated with a decreased risk of depression at 12 months postpartum (0.35, 95% CI: 0.19 to 0.63). Evidence suggests the recency of support also matters, but estimates are imprecise. We did not find evidence of a protective effect for support based on the MSSI. This study highlights the protective effect of sustained social support, particularly emotional support, on perinatal depression. Interventions targeting, leveraging and maintaining this type of support may be particularly important for reducing postpartum depression.


A proteomic approach to apoplastic proteins involved in cell wall regeneration in protoplasts of Arabidopsis suspension-cultured cells.
To clarify the mechanisms of cell wall construction, we used a proteomic approach to investigate the proteins secreted into cell wall spaces during cell wall regeneration from the protoplasts of Arabidopsis suspension-cultured cells. We focused on cell wall proteins loosely bound to the cell wall architecture and extractable with 1 M KCl solutions from: (i) native suspension cultured cells; (ii) protoplasts that had been allowed to regenerate their cell walls for 1 h; and (iii) protoplasts allowed to regenerate their cell walls for 3 h. We adopted a non-destructive extraction procedure without disrupting cellular integrity, thereby avoiding contamination from cytoplasmic proteins. Using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorption ionization-time-of-flight/mass spectrometry (MALDI-TOF/MS), we separated, mapped and identified 71 proteins derived from the native cell wall, and 175 and 212 proteins derived from the 1 and 3 h regenerated protoplasts, respectively. Quite different sets of proteins with differing status of their post-translational modifications, including phosphorylation and glycosylation, were identified in the three protein fractions. This indicated dynamic in muro changes in the cell wall proteins during cell wall regeneration in the protoplasts. The analysis revealed a set of enzymes specifically involved in cell wall expansion and construction in suspension-cultured cells. This approach has also determined a set of cell wall proteins that had not been predicted to be localized in cell wall spaces.


Human SP-A1 (SFTPA1) variant-specific 3' UTRs and poly(A) tail differentially affect the in vitro translation of a reporter gene.
Human surfactant protein A (SP-A) is encoded by two functional genes (SFTPA1, SFTPA2) with a high degree of sequence identity. Sequence differences among these genes and their genetic variants have been observed at the 5' and 3' untranslated regions (UTRs). In this work, we studied the impact on translation of the SFTPA1 (hSP-A1) and SFTPA2 (hSP-A2) gene 5' UTR splice variants and 3' UTR sequence variants, in the presence or absence of poly(A) tail. We generated constructs containing the luciferase reporter gene flanked upstream by one of the hSP-A 5' UTR splice variants and/or downstream by one hSP-A 3' UTR sequence variant. mRNA transcripts were prepared by in vitro transcription and used for either in vitro translation with a rabbit reticulocyte lysate or transient transfection of the lung adenocarcinoma cell line NCI-H441. The luciferase activity results indicate that hSP-A 5' UTR and 3' UTR together have an additive effect on translation. In this context, the hSP-A1 6A(3) and 6A(4) 3' UTR variants exhibited higher translation efficiency than the 6A(2) variant (P <0.05), whereas no significant difference was observed between the two hSP-A2 3' UTRs studied (1A(0), 1A(3)). Further sequence analysis revealed that a deletion of an 11-nucleotide (nt) element in both the 6A(3) and 6A(4) 3' UTR variants changes the predicted secondary structure stability and the number of putative miRNA binding sites. Removal of this 11-nt element in the 6A(2) 3' UTR resulted in increased translation, and the opposite effect was observed when the 11-nt element was cloned in a guest 3' UTR (6A(3), 6A(4)). These results indicate that sequence differences among hSP-A gene variants may account for differential regulation at the translational level.


Quantitative analysis of flux along the gluconeogenic, glycolytic and pentose phosphate pathways under reducing conditions in hepatocytes isolated from fed rats.
Hepatocytes were isolated from the livers of fed rats and incubated with a mixture of glucose (10 mM), ribose (1 mM), mannose (4 mM), glycerol (3 mM), acetate (1.25 mM), and ethanol (5 mM) with one substrate labelled with 14C in any given incubation. Incorporation of label into CO2, glucose, glycogen, lipid glycerol and fatty acids, acetate and C-1 of glucose was measured at 20 and 40 min after the start of the incubation. The data (about 48 measurements for each interval) were used in conjunction with a single-compartment model of the reactions of the gluconeogenic, glycolytic and pentose phosphate pathways and a simplified model of the relevant mitochondrial reactions. An improved method of computer analysis of the equations describing the flow of label through each carbon atom of each metabolite under steady-state conditions was used to compute values for the 34 independent flux parameters in this model. A good fit to the data was obtained, thereby permitting good estimates of most of the fluxes in the pathways under consideration. The data show that: net flux above the level of the triose phosphates is gluconeogenic; label in the hexose phosphates is fully equilibrated by the second 20 min interval; the triose phosphate isomerase step does not equilibrate label between the triose phosphates; substrate cycles are operating at the glucose-glucose 6-phosphate, fructose 6-phosphate-fructose 1,6-bisphosphate and phosphoenolpyruvate-pyruvate-oxaloacetate cycles; and, although net flux through the enzymes catalysing the non-oxidative steps of the pentose phosphate pathway is small, bidirectional fluxes are large.


[mRNA localization and content in bovine kininogen].
The bradykinin (BK) gene chemically synthesized and cloned into pBR 322 was used for the study of tissue localization and quantitation of mRNA coding for the BK precursor, kininogen. Poly(A+)-mRNAs from bovine liver, spleen, kidney, mammary gland and pancreas were used for dot-hybridization to [32P]DNA of the BK gene or to the plasmid-containing BK gene. The experimental results demonstrate that [32P]DNA of BK is hybridized only to the liver poly(A+)-RNA, which proves the liver to be the main kininogen mRNA-producing tissue. In other tissues, the kininogen mRNA synthesis is either altogether absent or its level is two orders of magnitude less as compared to the liver. Several approaches for the quantitation of the kininogen mRNA were developed. The amount of this mRNA was shown to be about 0.6% of the total cellular poly(A+)-RNA. Poly(A+)-RNA which is bound to BK DNA-cellulose and is enriched with BK-coding sequences was used for the study of the hybridization kinetics and translation in a cell-free system from rabbit reticulocytes. The polypeptides synthesized contain BK as was shown by the use of a bio-test in rat uterus.


Electrophysiologic transition from physiologic tremor to essential tremor.
We electrophysiologically examined the transition from physiologic tremor to essential tremor in people at risk for familial essential tremor. Two healthy people from different families with hereditary essential tremor were studied on multiple occasions. A 23-year-old man was studied in 1995, 1997, and 2004, and a 44-year-old woman was studied in 1993, 1995, 1997, and 2003. Hand acceleration and forearm electromyographic readings were measured with and without 300-g loading to determine the characteristic frequency-invariant motor unit entrainment of essential tremor. Clinically and electrophysiologically, the man and woman had normal tremor until the last examination, when both exhibited a fine tremulousness in the outstretched hands and frequency-invariant motor unit entrainment at 7.5 and 6.5 Hz, respectively. At no time did either patient exhibit a prominent 8-12 Hz component of physiologic tremor. Essential tremor in young adults may begin at frequencies less than 8-12 Hz, and this electrophysiologic abnormality is detectable when clinical examinations reveal only questionably abnormal tremor. More young adults at risk for essential tremor must be studied to determine whether initial frequencies less than 8 Hz are the rule or the exception. Nevertheless, the data from our 2 patients demonstrate that a prominent 8-12 Hz component of physiologic tremor does not always precede the development of essential tremor; therefore, the origins of essential tremor and the 8-12 Hz component of physiologic tremor may be different.


Confirmation of cross-reactivity between Lyme antibody H9724 and human heat shock protein 60 by a combinatorial approach.
A library of Ac-XXXXXPAPRM decapeptides was prepared on a TentaGel solid support using the approach of split synthesis and the strategy of partial chain termination. Epitope screening of the library (17(5) approximately 1.4 x 10(6) decapeptides) with a Lyme monoclonal antibody (H9724) and subsequent MALDI-MS analysis of candidate peptides from colored beads revealed a consensus structure of Xi-DLSXj (Xi = V, L, Y; Xj = G, R). These identified sequences presented no homology to the natural epitope from Borrelia burgdorferi flagellin. However, they were found to resemble a fragment at the N-terminus of human heat shock protein (Hsp60). Our results confirmed that H9724 cross-reacts between bacterial and human proteins and provided support for an autoimmunity mechanism of Lyme disease.


Physicians' Reactions to COVID-19: The Results of a Preliminary International Internet Survey.
Physicians across the world have been disproportionately affected by the COVID-19 pandemic. This study was designed and conducted to assess the emotional and behavioural reactions of physicians to the initial phase of the COVID-19 pandemic. An online survey questionnaire using the google forms platform was constructed by the authors. The items in the questionnaire were based on clinical experience, relevant literature review and discussion with peers. A list of issues that were deemed as essential components of the experience of the pandemic relevant to physicians was arrived at. Thereafter these issues were operationalized into question form and hosted on the google forms platform. The link to this questionnaire was circulated by the authors among their peer groups in the month of April 2020. We received 295 responses and 3 were unusable. Most of the responses were from India, the United States of America, Australia, Canada and the United Kingdom. About 60% of the respondents identified themselves as frontline and had a decade of clinical experience. Most respondents reported being anxious due to the pandemic and also observed the same in their peers and families. A majority also observed changes in behaviour in self and others and advanced a variety of reasons and concerns. A sense of duty was the most commonly employed coping mechanism. Physicians are not immune from information and misinformation, or cues in the environment. Behavioural choices are not always predicted by knowledge but by a combination of knowledge, emotional state, personality and environment. Healthcare settings need to be ready for emergencies and should focus on reducing uncertainty in physicians. These factors may also be gainfully used in the mental health promotion of physicians in COVID-19 care roles.


Translation termination in eukaryotes: from simplicity to complexity.
The last forty years of molecular biology and biochemistry were enormously rich in discoveries, which were not foreseen even by the most eminent scientists of that time. The findings of 1993-2000 profoundly enlarged our views on translation termination, clearly showing that our previous understanding was enormously oversimplified. Now the structural basis is created for much better insight into functions of termination factors. The story of translation termination in eukaryotes could be taken as an illustration of a general trend of molecular biology: "From simplicity to complexity". However, genuine knowledge requires that after this stage a third phase has to be reached, which is "From complexity to clarity". This has not yet been achieved in translation termination and therefore makes this topic quite attractive for researchers.


The diabetes healthy outcomes program: results of free health care for uninsured at a federally qualified community health center.
Uninsured patients with diabetes are at increased risk for poor outcomes and often have limited access to health and prescription services necessary to manage diabetes. Hamilton Health Center, a federally qualified community health center, with support from the Highmark Foundation, implemented a Diabetes Healthy Outcomes Program (DHOP) for uninsured patients. To evaluate the effectiveness of DHOP that is designed to provide health care and supportive services for uninsured diabetic patients at a federally qualified community health center. Mixed quantitative and qualitative analyses of participant outcomes and satisfaction were used to assess program effectiveness. A total of 189 participants enrolled in DHOP over 2 years. Thirty-four (18%) participants had adequate glycemic control with hemoglobin A1c (HbA1c) ≤ 7%. Overall, 105 participants received prescription drugs, 101 participants received eye care services, 23 participants received dental services, 45 received podiatry services, 37 received nutrition services, and 28 patients engaged in an exercise program. More participants (38%, 34) had controlled diabetes mellitus at study start than at the end (28%, 25). However, 30% versus 17% of participants with 2 HbA1c measurements achieved or maintained HbA1c ≤ 7% by the end of the program compared with the start. Participants who accessed more services were more likely to achieve glycemic control as measured by HbA1c (P > .01). Although 30% of participants improved or maintained glycemic control over 2 years, more were uncontrolled at the end than at study start. Participants who accessed more primary and specialty care services were more likely to achieve glycemic control. Multidisciplinary care may improve diabetes control in low-income patients.


Sequential sampling plans for the hairy chinch bug (Hemiptera: Lygaeidae).
Two sequential sampling plans were developed to produce tools to reduce the use of pesticides for the control of the hairy chinch bug, Blissus leucopterus hirtus Montandon, on turfgrass lawns in the cool-humid region of Quebec, Canada. A first plan based on Wald's method was shown to be too conservative in a validation conducted on infested lawns. The second plan developed using Iwao's method yielded good results. The difference between the two plans may have occurred because no common k could be found for the negative binomial functions describing chinch bug distribution on lawns, thereby violating an essential assumption of Wald's approach. Application of the Iwao plan, which is based on visual sampling of 0.1-m2 quadrats, requires approximately 20 min when lawn evaluation is conducted by a single person. Estimation of the occurrence of chinch bug infestations (28% infested lawns in the regions of Montreal and Quebec cities), and results from validation of the Iwao plan on infested lawns, indicate that the error rates alpha and beta of the plan are lower than an upper limit of 0.05 and 0.14, respectively. The adequate precision and practicality of this plan suggest that it could become an essential tool for management of turfgrass lawns in regions where the hairy chinch bug completes less than two generations per growing season.


Cell surface profiling using high-throughput flow cytometry: a platform for biomarker discovery and analysis of cellular heterogeneity.
Cell surface proteins have a wide range of biological functions, and are often used as lineage-specific markers. Antibodies that recognize cell surface antigens are widely used as research tools, diagnostic markers, and even therapeutic agents. The ability to obtain broad cell surface protein profiles would thus be of great value in a wide range of fields. There are however currently few available methods for high-throughput analysis of large numbers of cell surface proteins. We describe here a high-throughput flow cytometry (HT-FC) platform for rapid analysis of 363 cell surface antigens. Here we demonstrate that HT-FC provides reproducible results, and use the platform to identify cell surface antigens that are influenced by common cell preparation methods. We show that multiple populations within complex samples such as primary tumors can be simultaneously analyzed by co-staining of cells with lineage-specific antibodies, allowing unprecedented depth of analysis of heterogeneous cell populations. Furthermore, standard informatics methods can be used to visualize, cluster and downsample HT-FC data to reveal novel signatures and biomarkers. We show that the cell surface profile provides sufficient molecular information to classify samples from different cancers and tissue types into biologically relevant clusters using unsupervised hierarchical clustering. Finally, we describe the identification of a candidate lineage marker and its subsequent validation. In summary, HT-FC combines the advantages of a high-throughput screen with a detection method that is sensitive, quantitative, highly reproducible, and allows in-depth analysis of heterogeneous samples. The use of commercially available antibodies means that high quality reagents are immediately available for follow-up studies. HT-FC has a wide range of applications, including biomarker discovery, molecular classification of cancers, or identification of novel lineage specific or stem cell markers.


The Comparison of Immunoregulatory Properties of Human Alpha2-Macroglobulin and Pregnancy-Associated Alpha2-Glycoprotein.
Human alpha2-macroglobulin (MG) and pregnancy-associated alpha2-glycoprotein (PAG) are able to suppress proliferation of non-fractionated mononuclear blood cells and T- and B-enriched populations in spontaneous and mitogen-stimulated cultures in vitro. Immunosuppressive properties of both proteins are more expressed in mitogen-stimulated cultures than in spontaneous ones, and in T population than in B one. PAG is a more powerful immunosuppressor, because its suppressive effect is more than ten-fold greater than corresponding effect of MG. Female peripheral blood mononuclear cells are more susceptible to immunoregulatory effect of alpha2-glycoprotein than male ones. This fact may confirm the known alpha2-glycoprotein role as an immunoregulator in pregnancy.


The effects of colorectally insufflated oxygen-ozone on red blood cell rheology in rabbits.
Currently, with reappraisal of ozone therapy, it has been utilized worldwide in research and clinical field. Most of the studies investigating effects of ozone on blood parameters are conducted by directly ozonating the blood. Rectal insufflation is a simple, easy and inexpensive method of delivering ozone. Little is known how these gases affect some fundamental hemorheologic parameters when given by insufflation. We aimed to investigate the effects of colorectally insufflated oxygen-ozone on red blood cell rheology in rabbits. Rabbits were divided into Group 1 (control); Groups 2, 3 and 4 (oxygen rectally insufflated respectively for 15, 21 and 36 days); Groups 5, 6 and 7 (ozone rectally insufflated respectively for 15, 21 and 36 days). Erythrocyte deformability, aggregation and osmotic fragility were determined from blood samples at the end of each treatment period. Our study showed an improvement in deformability, a decrease in aggregation and an increase in fragility following a 15 day ozone treatment. With longer ozone application the changes in aggregation and fragility returned back to control levels, however its effect on deformability sustained. Therefore, more than two weeks ozone insufflation may induce adaptation to changes induced by ozone suggesting its systemic effects.


Observation of the synchronization of chaos in mutually injected vertical-cavity surface-emitting semiconductor lasers.
Synchronization of chaotic oscillations was observed in mutually injected vertical-cavity surface-emitting lasers (VCSELs) in a low-frequency fluctuation regime. In the experiments, only one of the two polarization modes (x mode) showed synchronized oscillations, and the other polarization components (y mode) were synchronized as a result of the effect of anticorrelated oscillations that is a characteristic feature of VCSELs.


Infection in exacerbations of asthma: views of different groups of practitioners.
A questionnaire designed to elucidate views on the frequency of infection in asthma and its management was circulated to general practitioners, general physicians, paediatricians and respiratory physicians in the north of England. On the whole, general practitioners agreed with the general physicians and paediatricians with respiratory physicians. The generalists, and to a lesser extent the specialists, tended to overestimate the risk of bacterial infection, which was perceived to be a particular risk following viral infection in intrinsic asthma. This resulted in more frequent prescription of antibiotics than is justified by the published evidence. There was a tendency for frequent prescribers of antibiotics to withhold corticosteroids. If prescribing habits are to be altered, education will have to be directed at those responsible for acute admissions to hospital as well as at general practitioners.


Highly stable noble-metal nanoparticles in tetraalkylphosphonium ionic liquids for in situ catalysis.
Gold and palladium nanoparticles were prepared by lithium borohydride reduction of the metal salt precursors in tetraalkylphosphonium halide ionic liquids in the absence of any organic solvents or external nanoparticle stabilizers. These colloidal suspensions remained stable and showed no nanoparticle agglomeration over many months. A combination of electrostatic interactions between the coordinatively unsaturated metal nanoparticle surface and the ionic-liquid anions, bolstered by steric protection offered by the bulky alkylated phosphonium cations, is likely to be the reason behind such stabilization. The halide anion strongly absorbs to the nanoparticle surface, leading to exceptional nanoparticle stability in halide ionic liquids; other tetraalkylphosphonium ionic liquids with non-coordinating anions, such as tosylate and hexafluorophosphate, show considerably lower affinities towards the stabilization of nanoparticles. Palladium nanoparticles stabilized in the tetraalkylphosphonium halide ionic liquid were stable, efficient, and recyclable catalysts for a variety of hydrogenation reactions at ambient pressures with sustained activity. Aerial oxidation of the metal nanoparticles occurred over time and was readily reversed by re-reduction of oxidized metal salts.


Naturally occurring mutation affecting the MyD88-binding site of TNFRSF13B impairs triggering of class switch recombination.
Mutations in the transmembrane activator and calcium-modulating cyclophilin ligand interactor (TACI) were previously found to be associated with hypogammaglobulinemia in humans. It has been shown that proliferation inducing ligand (APRIL) elicits class switch recombination (CSR) by inducing recruitment of MyD88 to a TACI highly conserved cytoplasmic domain (THC). We have identified a patient with hypogammaglobulinemia carrying a missense mutation (S231R) predicted to affect the THC. Aiming to evaluate the relevance of this novel mutation of TACI in CSR induction, we tested the ability of TACI, TLR9, or/and CD40 ligands to trigger CSR in naive B cells and B-cell lines carrying S231R. IgG secretion was impaired when triggered by TACI or/and TLR9 ligands on S231R-naive B cells. Likewise, these stimuli induced less expression of activation-induced cytidine deaminase, I(γ)1-C(μ), and I(γ)1-C(μ), while induction by optimal CD40 stimulation was indistinguishable from controls. These cells also showed an impaired cooperation between TACI and TLR9 pathways, as well as a lack of APRIL-mediated enhancement of CD40 activation in suboptimal conditions. Finally, after APRIL ligation, S231R-mutated TACI failed to colocalize with MyD88. Collectively, these results highlight the requirement of an intact MyD88-binding site in TACI to trigger CSR.


Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock.
We performed a comparative analysis of the Vibrio cholerae strain El Tor 3083 entering the viable but non-culturable (VBNC) state and starvation after incubation in artificial seawater (ASW) at 4 and 15 degrees C respectively. To this end, we determined bacterial culturability and membrane integrity, as well as the cellular levels of 16S rRNA and mRNA for the tuf, rpoS and relA genes, which were assessed by real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). Bacterial cells entering the VBNC state showed a 154, 5.1 x 10(3), 24- and 23-fold reduction in the number of copies of 16S rRNA and mRNA for tuf, rpoS and relA, in comparison to exponentially growing cells. The differences were less striking between cells in the VBNC and starvation states. The mRNA for relA was selectively increased in VBNC cells (3.2-folds), whereas a 3.9-fold reduction was observed for 16S rRNA. The obtained results confirmed that key activities of the cellular metabolism (i.e. tuf representing protein synthesis, and relA or rpoS stress response) were still detected in bacteria entering the VBNC state and starvation. These data suggest that the new Q-RT-PCR methodology, based on the selected RNA targets, could be successfully exploited for the identification (rRNA) of V. cholerae and assessment of its metabolic activity (tuf, rpoS, relA mRNA) in environmental samples.


Nature of the intense near-IR absorption and unusual broad UV-visible-NIR spectra of azulenocyanines: density functional theory studies.
The nature of the broad and intense near-IR (NIR) absorptions of azulenocyanine (H(2)Azc) in the range of 720-1300 nm due to the π→π* transitions is clearly revealed on the basis of density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations. The electron moving between peripheral conjugated systems and the eighteen electron-π-conjugated core due to different types of electron transitions has also been clarified. Because of the small gap between the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) of all four H(2)Azc isomers, their Q absorption bands in the region of 850-1300 nm are significantly red-shifted relative to its isoelectronic analogue naphthalocyanine H(2)Nc. Observation of the new absorption bands between Q and Soret bands from 720 to 1050 nm for H(2)Azc is considered to be the result of electron transitions from the fused azulene rings to the 18 electron-π-conjugated core. In addition, the introduction of electron-withdrawing or electron-donating substituents onto the periphery of the azulenocyanine ring is revealed to further reduce the LUMO-HOMO gap, leading to a further red-shifted broad and intense NIR absorption band from 900 to 1700 nm for H(2)(F(8)Azc) and H(2)[(NH(2))(8)Azc].


Connective tissue response to immobility. Correlative study of biomechanical and biochemical measurements of normal and immobilized rabbit knees.
The biomechanical changes in rabbit knee contractures were measured after 9 weeks of immobilization. These results were correlated with the biochemical composition of periarticular connective knee tissue of the same knees. The loss of total hexosamine correlates significantly with joint stiffness on an animal-to-animal basis. Total hexosamine also correlates with the energy required cyclicly to flex and to extend the experimental joints. Of the individual glycosaminoglycan fractions, only hyaluronic acid showed significant correlation with the biomechanical data. The possible mechanism of contracture formation was postulated based on these results.


A murine model of peanut anaphylaxis: T- and B-cell responses to a major peanut allergen mimic human responses.
Peanut allergy affects 0.6% of the US population. At the present time, allergen avoidance is the only therapeutic option. Animal models of food-induced anaphylaxis would facilitate attempts to design novel immunotherapeutic strategies for the treatment of peanut allergy. The purpose of this study was to develop a murine model of IgE-mediated peanut hypersensitivity that closely mimics human peanut allergy. C3H/HeJ mice sensitized orally with freshly ground whole peanut and cholera toxin as adjuvant were challenged orally 3 and 5 weeks later with crude peanut extract. Anaphylactic reactions were determined. T- and B-cell responses to Ara h 1 and Ara h 2, the major peanut allergens, were characterized by evaluating splenocyte proliferative responses and IgE antibody concentrations. Furthermore, IgE antibodies in the sera of patients with peanut allergy and mice were compared for antibody binding to Ara h 2 isoforms and allergenic epitopes. Peanut-specific IgE was induced by oral peanut sensitization, and hypersensitivity reactions were provoked by feeding peanut to sensitized mice. The symptoms were similar to those seen in human subjects. Ara h 1- and Ara h 2-specific antibodies were present in the sera of mice with peanut allergy. Furthermore, these Ara h 2-specific IgE antibodies bound the same Ara h 2 isoforms and major allergenic epitopes as antibodies in the sera of human subjects with peanut allergy. Splenocytes from mice with peanut allergy exhibited proliferative responses to Ara h 1 and Ara h 2. This murine model of peanut allergy mimics the clinical and immunologic characteristics of peanut allergy in human subjects and should be a useful tool for developing immunotherapeutic approaches for the treatment of peanut allergy.


Peptidylarginine deiminase 4 and citrullination in health and disease.
Deimination is catalyzed by a family of calcium binding enzymes, called peptidylarginine deiminases (PADs). Among these, the PAD4 isoform has been more extensively studied for its role in some autoimmune diseases. PAD4 is localized in the cytoplasm of monocytes, T and B cells, neutrophils, eosinophils and NK cells and can move to the nucleus upon cell activation. PAD4 plays a physiological role in gene regulation via citrullination of histones. In rheumatoid arthritis (RA), PAD4 contributes to the generation of ACPA specific substrates and is itself a target of autoantibodies; alleles of the PADI4 gene confer susceptibility to RA in Asians but not in Caucasians. In multiple sclerosis, extensive deimination of brain proteins is observed in active lesions, but no role for the PADI4 gene in susceptibility to MS has been so far described.


Strengths-Based Assessment for Suicide Prevention: Reasons for Life as a Protective Factor From Yup'ik Alaska Native Youth Suicide.
Suicide is the second leading cause of death among American Indian and Alaska Native youth, and within the Alaska Native youth subpopulation, the leading cause of death. In response to this public health crisis, American Indian and Alaska Native communities have created strategies to protect their young people by building resilience using localized Indigenous well-being frameworks and cultural strengths. These approaches to suicide prevention emphasize promotion of protective factors over risk reduction. A measure of culturally based protective factors from suicide risk has potential to assess outcomes from these strengths-based, culturally grounded suicide prevention efforts, and can potentially address several substantive concerns regarding direct assessment of suicide risk. We report on the Reasons for Life (RFL) scale, a measure of protective factors from suicide, testing psychometric properties including internal structure with 302 rural Alaska Native Yup'ik youth. Confirmatory factor analyses revealed the RFL is best described through three distinct first-order factors organized under one higher second-order factor. Item response theory analyses identified 11 satisfactorily functioning items. The RFL correlates with other measures of more general protective factors. Implications of these findings are described, including generalizability to other American Indian and Alaska Native, other Indigenous, and other culturally distinct suicide disparities groups.


Diversity of translation start sites may define increased complexity of the human short ORFeome.
Our previous proteomics analysis of small proteins expressed in human K562 cells provided the first direct evidence of translation of upstream ORFs in human full-length cDNAs (Oyama, M., Itagaki, C., Hata, H., Suzuki, Y., Izumi, T., Natsume, T., Isobe, T., and Sugano, S. (2004) Analysis of small human proteins reveals the translation of upstream open reading frames of mRNAs. Genome Res. 14, 2048-2052). In the present study, we performed an in-depth proteomics analysis of human K562 and HEK293 cells using a two-dimensional nano-liquid chromatography-tandem mass spectrometry system. The results led to the identification of eight protein-coding regions besides 197 small proteins with a theoretical mass less than 20 kDa that were already annotated coding sequences in the curated mRNA database. In addition to the upstream ORFs in the presumed 5'-untranslated regions of mRNAs, bioinformatics analysis based on accumulated 5'-end cDNA sequence data provided evidence of novel short coding regions that were likely to be translated from the upstream non-AUG start site or from the new short transcript variants generated by utilization of downstream alternative promoters. Protein expression analysis of the GRINL1A gene revealed that translation from the most upstream start site occurred on the minor alternative splicing transcript, whereas this initiation site was not utilized on the major mRNA, resulting in translation of the downstream ORF from the second initiation codon. These findings reveal a novel post-transcriptional system that can augment the human proteome via the alternative use of diverse translation start sites coupled with transcriptional regulation through alternative promoters or splicing, leading to increased complexity of short protein-coding regions defined by the human transcriptome.


The calcium-sensing receptor in human disease.
The discovery of the calcium-sensing receptor (CaR), a G protein-coupled receptor, has led to the elucidation of the pivotal roles of the CaR in systemic calcium homeostasis. The receptor is situated on the chief cells of the parathyroid glands, where it senses the extracellular Ca2+ concentration and in turn alters the rate of secretion of parathyroid hormone (PTH). The intracellular signal pathways to which the CaR couples include, but are not limited to, phospholipase C (PLC), and mitogen-activated protein kinases. The receptor is widely expressed in various tissues and likely serves important cellular functions beyond that of maintaining systemic calcium homeostasis. Functionally important mutations in the receptor have been found to cause disorders in calcium homeostasis due both to changes in the set point for PTH secretion and to the control of renal calcium excretion. These mutations cause hypercalcemia when the mutation inactivates the receptor and cause hypocalcemia when the mutation activates the receptor. Recent studies have revealed the presence of circulating autoantibodies to the calcium-sensing receptor in humans, with the clinical presentation the same as that for diseases caused by mutations in the CaR. In renal secondary hyperparathyroidism, a drug that stimulates the receptor (calcimimetic) shows great promise as a medical treatment for this condition.


All-Atom MD Simulation of DNA Condensation Using Ab Initio Derived Force Field Parameters of Cobalt(III)-Hexammine.
It is well established that the presence of the trivalent cobalt(III)-hexammine cation (CoHex3+) at submillimolar concentrations leads to bundling (condensation) of double-stranded DNA molecules, which is caused by DNA-DNA attraction induced by the multivalent counterions. However, the detailed mechanism of this process is still not fully understood. Furthermore, in all-atom molecular dynamics (MD) simulations, spontaneous aggregation of several DNA oligonucleotides in the presence of CoHex3+ has previously not been demonstrated. In order to obtain a rigorous description of CoHex3+-nucleic acid interactions and CoHex3+-induced DNA condensation to be used in MD simulations, we have derived optimized force field parameters of the CoHex3+ ion. They were obtained from Car-Parrinello molecular dynamics simulation of a single CoHex3+ ion in the presence of 125 water molecules. The new set of force field parameters reproduces the experimentally known transition of DNA from B- to A-form, and qualitatively describes changes of DNA and RNA persistence lengths. We then carried out a 2 μs long atomistic simulation of four DNA oligomers each consisting of 36 base pairs in the presence of CoHex3+. We demonstrate that, in this system, DNA molecules display attractive interactions and aggregate into bundle-like structures. This behavior depends critically on the details of the CoHex3+ interaction with DNA. A control simulation with a similar setup but in the presence of Mg2+ does not induce DNA-DNA attraction, which is also in agreement with experiment.


The trauma model of dissociation: inconvenient truths and stubborn fictions. Comment on Dalenberg et al. (2012).
Dalenberg et al. (2012) argued that convincing evidence (a) supports the longstanding trauma model (TM), which posits that early trauma plays a key role in the genesis of dissociation; and (b) refutes the fantasy model (FM), which posits that fantasy proneness, suggestibility, cognitive failures, and other variables foster dissociation. We review evidence bearing on Dalenberg et al.'s 8 predictions and find them largely wanting in empirical support. We contend that the authors repeat errors committed by many previous proponents of the TM, such as attributing a central etiological role to trauma in the absence of sufficient evidence. Specifically, Dalenberg et al. leap too quickly from correlational data to causal conclusions, do not adequately consider the lack of corroboration of abuse in many studies, and underestimate the relation between dissociation and false memories. Nevertheless, we identify points of agreement between the TM and FM regarding potential moderators and mediators of dissociative symptoms (e.g., family environment, biological vulnerabilities) and the hypothesis that dissociative identity disorder is a disorder of self-understanding. We acknowledge that trauma may play a causal role in dissociation but that this role is less central and specific than Dalenberg et al. contend. Finally, although a key assumption of the TM is dissociative amnesia, the notion that people can encode traumatic experiences without being able to recall them lacks strong empirical support. Accordingly, we conclude that the field should now abandon the simple trauma-dissociation model and embrace multifactorial models that accommodate the diversity of causes of dissociation and dissociative disorders.


Adverse birth outcomes among native-born and immigrant women: replicating national evidence regarding Mexicans at the local level.
For almost two decades, the literature has consistently described an epidemiologic paradox relating to better birth outcomes among high-risk groups, particularly new immigrants from Mexico and Southeast Asia. We hypothesize that regardless of their sociodemographic profile, Mexican immigrants will exhibit lower rates of low birth weight and preterm deliveries than native-(U.S.) born women of Mexican origin, non-Hispanic White and Black women, and Puerto Rican Women. We studied 57,324 live-born singleton infants born to residents in the city of Chicago in a linked data set of 1994 birth-death records. Multivariate logistic regression was used to analyze race/ethnicity differentials in two pregnancy outcome measures, low birth weight and preterm birth. Overall better birth outcome is related to maternal immigrant status regardless of race/ethnic groups. Immigrant Mexican women had a significantly lower risk of both low birth weight [adjusted odds ratio (AOR): 0.78, 95% confidence interval (CI) 0.66-0.91] and preterm births (AOR: 0.75, 95% CI 0.65-0.86) and were at 28% and 33% lower risks of delivering a low birth weight infant or a premature infant, respectively, than non-Hispanic White women.


Corticosteroid-associated tendinopathies: an analysis of the published literature and spontaneous pharmacovigilance data.
Corticosteroids may cause tendinopathies, an adverse effect that is not well known and characterised, although it was initially described more than 40 years ago. This study was conducted in order to characterise the important aspects of this adverse reaction, such as the role played by routes of corticosteroid administration, therapy duration, comedication, cumulative corticosteroid dose and underlying disease. Published case reports of tendinopathies that were associated with corticosteroid use were identified by a comprehensive literature search using the databases MEDLINE, Pharm-line, EMBASE, ToxFile, Adis Inpharma, International Pharmaceutical Abstracts, Drug Information Fulltext and PASCAL. The reference lists of all pertinent articles were cross-referenced to retrieve additional cases. Spontaneous reports were requested from the Uppsala Monitoring Centre (the WHO Collaborating Centre for International Drug Monitoring). Information of published and spontaneous reports was analysed with regard to age, sex, underlying disease, individual corticosteroids, equivalent corticosteroid dose, latency time, cumulative dose, route of administration, comedication and type of tendinopathy. We included and analysed 73 published case reports and case series involving 133 patients and 191 spontaneous reports of corticosteroid-associated tendinopathies. The proportion of women with tendinopathies was 50.8% and 41.0%, respectively. The mean age (+/- standard deviation) of patients with a tendinopathy was 50 +/- 17 years and 61 +/- 16 years, respectively. The predominant routes of administration were oral (33% of published cases and 47% of spontaneous cases) and intra-articular (35% of the published cases or parenteral (7% of the spontaneous cases). There were isolated cases of tendinopathy after inhaled or topical (i.e. ocular, cutaneous, nasal) corticosteroid use. Tendinopathies that were reported in the literature cases consisted mainly of tendon ruptures (93%), predominantly of the Achilles tendon. Of the spontaneous cases, 31% had a rupture and the remaining cases had a tendinitis or an unspecified tendon disorder. Oral and parenteral applications, especially intra-articular use, were the most prevalent routes of administration in cases with corticosteroid-associated tendinopathies. However, topical application has also been rarely associated with tendinopathies. Future pharmacoepidemiological studies should further address this issue to quantify the risk of corticosteroid-associated tendinopathies.


Shape tailoring of AgBr microstructures: effect of the cations of different bromide sources and applied surfactants.
Investigations regarding AgBr-based photocatalysts came to the center of attention due to their high photosensitivity. The present research focuses on the systematic investigation regarding the effect of different alkali metal cation radii and surfactants/capping agents applied during the synthesis of silver-halides. Their morpho-structural and optical properties were determined via X-ray diffractometry, diffuse reflectance spectroscopy, scanning electron microscopy, infrared spectroscopy, and contact angle measurements. The semiconductors' photocatalytic activities were investigated using methyl orange as the model contaminant under visible light irradiation. The correlation between the photocatalytic activity and the obtained optical and morpho-structural properties was analyzed using generalized linear models. Moreover, since the (photo)stability of Ag-based photoactive materials is a crucial issue, the stability of catalysts was also investigated after the degradation process. It was concluded that (i) the photoactivity of the samples could be fine-tuned using different precursors and surfactants, (ii) the as-obtained AgBr microcrystals were transformed into other Ag-containing composites during/after the degradation, and (iii) elemental bromide did not form during the degradation process. Thus, the proposed mechanisms in the literature (for the degradation of MO using AgBr) must be reconsidered.


A simple liquid chromatography method running in dual modes for quantification of short and medium chain fatty acids.
Sample preparation is often a rate-limiting step in quantification of short chain and medium chain fatty acids (SFAs and MFAs) in biological samples. A novel liquid chromatography (LC) method with simple sample preparation is introduced in this report. The method is performed on a column packed with polyvinyl alcohol sorbents. In separation of a mixture of bovine serum albumin (BSA) with SFAs and MFAs, the protein is eluted in size-exclusion mode in the first part of the chromatogram. SFAs and MFAs are then eluted in the second part of the chromatogram in reverse-phase mode where fatty acids with longer carbon lengths are eluted at longer retention times. SFAs and MFAs are well resolved from each other without pre-column derivatization. The size-exclusion mode affords good tolerance to macromolecules in sample matrixes. The dual mode separation allows sample injections without complicated sample preparation steps, such as derivatization, extraction, evaporation, and reconstitution.


Fitness costs of Vip3A resistance in Spodoptera frugiperda on different hosts.
Fall armyworm, Spodoptera frugiperda, is a major target pest of the Vip3A protein. The fitness of Bacillus thuringiensis (Bt)-resistant insects on refuges is an important factor in determining the speed of resistance development. Fitness costs associated with Bt resistance can vary among host plants. Here, we provide the first experimental evaluation of the interactions between non-Bt hosts and fitness costs of Vip3A resistance in S. frugiperda. Laboratory bioassays showed that survivorship, pupal weight, sex ratio, developmental time, fecundity, net reproductive rate, and intrinsic rate were not different among Vip3A-resistant (RR), -susceptible (SS) and -heterozygous (R♂ S♀ and R♀ S♂ ) strains of S. frugiperda on non-Bt corn, non-Bt cotton, and non-Bt diet. R♂ S♀ and R♀ S♂ also showed no differences on sorghum relative to SS. However, compared to SS, RR on sorghum showed significant reduction in pupal weight, and took longer time to develop to pupa and adult. These findings suggest that recessive fitness costs of reduced pupal weight and growth rate were detected at the individual level in the Vip3A resistant populations of S. frugiperda on sorghum. However, at the population level in terms of net reproductive rate and intrinsic rate, fitness costs of Vip3A resistance was not evident in S. frugiperda. © 2018 Society of Chemical Industry.


B cell MHC haplotype affects follicular inclusion, germinal center participation and plasma cell differentiation in a mouse model of lupus.
MHC class II molecules are essential for appropriate immune responses against pathogens but are also implicated in pathological responses in autoimmune diseases and transplant rejection. Previous studies have shed light on the systemic contributions of MHC haplotypes to the development and severity of autoimmune diseases. In this study, we addressed the B cell intrinsic MHC haplotype impact on follicular inclusion, germinal center (GC) participation and plasma cell (PC) differentiation in the context of systemic lupus erythematosus (SLE). We leveraged the 564Igi mouse model which harbors a B cell receptor knock-in from an autoreactive B cell clone recognizing ribonuclear components, including double-stranded DNA (dsDNA). This model recapitulates the central hallmarks of the early stages of SLE. We compared 564Igi heterozygous offspring on either H2b/b, H2b/d, or H2d/d background. This revealed significantly higher germinal center (GC) B cell levels in the spleens of H2b/b and H2b/d as compared to H2d/d (p<0.0001) mice. In agreement with this, anti-dsDNA-antibody levels were higher in H2b/b and H2b/d than in H2d/d (p<0.0001), with H2b/b also being higher compared to H2b/d (p<0.01). Specifically, these differences held true both for autoantibodies derived from the knock-in clone and from wild-type (WT) derived clones. In mixed chimeras where 564Igi H2b/b, H2b/d and H2d/d cells competed head-to-head in the same environment, we observed a significantly higher inclusion of H2b/b cells in GC and PC compartments relative to their representation in the B cell repertoire, compared to H2b/d and H2d/d cells. Furthermore, in mixed chimeras in which WT H2b/b and WT H2d/d cells competed for inclusion in GCs associated with an epitope spreading process, H2b/b cells participated to a greater extent and contributed more robustly to the PC compartment. Finally, immature WT H2b/b cells had a higher baseline of BCRs with an autoreactive idiotype and were subject to more stringent negative selection at the transitional stage. Taken together, our findings demonstrate that B cell intrinsic MHC haplotype governs their capacity for participation in the autoreactive response at multiple levels: follicular inclusion, GC participation, and PC output. These findings pinpoint B cells as central contributors to precipitation of autoimmunity.


Combination of orthodontic movement and periodontal therapy for full root coverage in a Miller class III recession: a case report with 12 years of follow-up.
One of the main purposes of mucogingival therapy is to obtain full root coverage. Several treatment modalities have been developed, but few techniques can provide complete root coverage in a class III Miller recession. Thus, the aim of this case report is to present a successful clinical case of a Miller class III gingival recession in which complete root coverage was obtained by means of a multidisciplinary approach. A 17-year-old Caucasian female was referred for treatment of a gingival recession on the mandibular left central incisor. The following procedures were planned for root coverage in this case: free gingival graft, orthodontic movement by means of alignment and leveling and coronally advanced flap (CAF). The case has been followed up for 12 years and the patient presents no recession, no abnormal probing depth and no bleeding on probing, with a wide attached gingiva band. A compromised tooth with poor prognosis, which would be indicated for extraction, can be treated by orthodontic movement and periodontal therapy, with possibility of 100% root coverage in some class III recessions.


Protein kinase C-δ signaling regulates glucagon secretion from pancreatic islets.
Accumulating evidence supports the "glucagonocentric hypothesis", in which antecedent α-cell failure and inhibition of glucagon secretion are responsible for diabetes progression. Protein kinase C (PKC) is involved in glucagon secretion from α-cells, although which PKC isozyme is involved and the mechanism underlying this PKC-regulated glucagon secretion remains unknown. Here, the involvement of PKCδ in the onset and progression of diabetes was elucidated. Immunofluorescence studies revealed that PKCδ was expressed and activated in α-cells of STZ-induced diabetic model mice. Phorbol 12-myristate 13-acetate (PMA) stimulation significantly augmented glucagon secretion from isolated islets. Pre-treatment with quercetin and rottlerin, PKCδ signaling inhibitors, significantly suppressed the PMA-induced elevation of glucagon secretion. While Go6976, a Ca2+-dependent PKC selective inhibitor did not suppress glucagon secretion. Quercetin suppressed PMA-induced phosphorylation of Tyr311 of PKCδ in isolated islets. However, quercetin itself had no effect on either glucagon secretion or glucagon mRNA expression. Our data suggest that PKCδ signaling inhibitors suppressed glucagon secretion. Elucidation of detailed signaling pathways causing PKCδ activation in the onset and progression of diabetes followed by the augmentation of glucagon secretion could lead to the identification of novel therapeutic target molecules and the development of novel therapeutic drugs for diabetes. J. Med. Invest. 64: 122-128, February, 2017.


Septic Arthritis of the Pediatric Shoulder: From Infancy to Adolescence.
Background. Septic arthritis of the glenohumeral joint in the pediatric population (<18 yo) is not commonly described in the literature. There is a corresponding paucity of information regarding its presentation and treatment. Methods. An IRB approved review of patients treated with irrigation and debridement by the Orthopaedic Department was completed. This retrospective study includes four patients, presenting from 2005 to 2015, with septic arthritis of the shoulder. Results. The mean age (M age) at presentation was 5 years, with a range of 1 month to 15 years. Patients presented on average after 7 days with pain and a mean temperature of 39°C, erythrocyte sedimentation rate of 66 mm/hr, a C-reactive protein level of 11.17 g/dL, and a white blood cell count of 20.2 × 10(3)/mcL. Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa were cultured from the wounds. All cases were treated operatively with irrigation and debridement and with antimicrobial therapy. Patients received antibiotics for an average of 6 weeks. Conclusion. Septic arthritis of the shoulder occurs in all pediatric ages. Successful treatment of septic arthritis of the shoulder was accomplished in four cases without division of the biceps sheath, with an average follow-up of 8 months.


Mental health and quality-of-life concerns related to the burden of food allergy.
As food allergy increases, more research is devoted to its influence on patient and family mental health and quality of life (QoL). This article discusses the effects on parent and child QoL, as well as distress, while appraising the limitations of knowledge given the methods used. Topics include whether QoL and distress are affected compared with other illnesses, assessment of distress and QoL in parents compared with children, concerns about food allergy-related bullying, and the necessity for evidence-based interventions. Suggestions are offered for how to improve QoL and reduce distress on the way to better coping with food allergy.


Weight gain in smokers after quitting cigarettes: meta-analysis.
To describe weight gain and its variation in smokers who achieve prolonged abstinence for up to 12 months and who quit without treatment or use drugs to assist cessation. Meta-analysis. We searched the Central Register of Controlled Trials (CENTRAL) and trials listed in Cochrane reviews of smoking cessation interventions (nicotine replacement therapy, nicotinic partial agonists, antidepressants, and exercise) for randomised trials of first line treatments (nicotine replacement therapy, bupropion, and varenicline) and exercise that reported weight change. We also searched CENTRAL for trials of interventions for weight gain after cessation. Trials were included if they recorded weight change from baseline to follow-up in abstinent smokers. We used a random effects inverse variance model to calculate the mean and 95% confidence intervals and the mean of the standard deviation for weight change from baseline to one, two, three, six, and 12 months after quitting. We explored subgroup differences using random effects meta-regression. 62 studies were included. In untreated quitters, mean weight gain was 1.12 kg (95% confidence interval 0.76 to 1.47), 2.26 kg (1.98 to 2.54), 2.85 kg (2.42 to 3.28), 4.23 kg (3.69 to 4.77), and 4.67 kg (3.96 to 5.38) at one, two, three, six, and 12 months after quitting, respectively. Using the means and weighted standard deviations, we calculated that at 12 months after cessation, 16%, 37%, 34%, and 13% of untreated quitters lost weight, and gained less than 5 kg, gained 5-10 kg, and gained more than 10 kg, respectively. Estimates of weight gain were similar for people using different pharmacotherapies to support cessation. Estimates were also similar between people especially concerned about weight gain and those not concerned. Smoking cessation is associated with a mean increase of 4-5 kg in body weight after 12 months of abstinence, and most weight gain occurs within three months of quitting. Variation in weight change is large, with about 16% of quitters losing weight and 13% gaining more than 10 kg.


Reverse engineering of force integration during mitosis in the Drosophila embryo.
The mitotic spindle is a complex macromolecular machine that coordinates accurate chromosome segregation. The spindle accomplishes its function using forces generated by microtubules (MTs) and multiple molecular motors, but how these forces are integrated remains unclear, since the temporal activation profiles and the mechanical characteristics of the relevant motors are largely unknown. Here, we developed a computational search algorithm that uses experimental measurements to 'reverse engineer' molecular mechanical machines. Our algorithm uses measurements of length time series for wild-type and experimentally perturbed spindles to identify mechanistic models for coordination of the mitotic force generators in Drosophila embryo spindles. The search eliminated thousands of possible models and identified six distinct strategies for MT-motor integration that agree with available data. Many features of these six predicted strategies are conserved, including a persistent kinesin-5-driven sliding filament mechanism combined with the anaphase B-specific inhibition of a kinesin-13 MT depolymerase on spindle poles. Such conserved features allow predictions of force-velocity characteristics and activation-deactivation profiles of key mitotic motors. Identified differences among the six predicted strategies regarding the mechanisms of prometaphase and anaphase spindle elongation suggest future experiments.


Ca2+ administration prevents α-synuclein proteotoxicity by stimulating calcineurin-dependent lysosomal proteolysis.
The capacity of a cell to maintain proteostasis progressively declines during aging. Virtually all age-associated neurodegenerative disorders associated with aggregation of neurotoxic proteins are linked to defects in the cellular proteostasis network, including insufficient lysosomal hydrolysis. Here, we report that proteotoxicity in yeast and Drosophila models for Parkinson's disease can be prevented by increasing the bioavailability of Ca2+, which adjusts intracellular Ca2+ handling and boosts lysosomal proteolysis. Heterologous expression of human α-synuclein (αSyn), a protein critically linked to Parkinson's disease, selectively increases total cellular Ca2+ content, while the levels of manganese and iron remain unchanged. Disrupted Ca2+ homeostasis results in inhibition of the lysosomal protease cathepsin D and triggers premature cellular and organismal death. External administration of Ca2+ reduces αSyn oligomerization, stimulates cathepsin D activity and in consequence restores survival, which critically depends on the Ca2+/calmodulin-dependent phosphatase calcineurin. In flies, increasing the availability of Ca2+ discloses a neuroprotective role of αSyn upon manganese overload. In sum, we establish a molecular interplay between cathepsin D and calcineurin that can be activated by Ca2+ administration to counteract αSyn proteotoxicity.


Pulmonary Rehabilitation Improves Self-Management Ability in Subjects With Obstructive Lung Disease.
Optimizing self-management is a key element in multidisciplinary pulmonary rehabilitation in patients with asthma or COPD. This observational study aimed to investigate the changes in self-management following pulmonary rehabilitation in subjects with chronic lung disease. Data were prospectively and routinely gathered at initial assessment and discharge in subjects taking part in a 12-week multidisciplinary out-patient pulmonary rehabilitation program. Measures of self-management included the Patient Activation Measure (PAM), the Health Education Impact Questionnaire (HEIQ) (8 subscales), a Self-Efficacy Questionnaire (2 subscales), the Lung Information Needs Questionnaire (LINQ), and the Health Literacy Questionnaire (HLQ) (9 subscales). Mean differences with 95% CI and effect sizes were computed. A total of 70 subjects (62.9% women) were included, with a median age of 63.5 y; most of the subjects had been diagnosed with COPD (77%). Between admission and discharge, all measures of self-management increased significantly except for the HEIQ subscales of constructive attitudes and approaches, social integration and support, and health services navigation; and the HLQ subscale of social support for health. The largest improvements (effect size > 0.55) were seen for the PAM (0.57); the HEIQ subscales of health-directed behavior (0.71), self-monitoring and insight (0.62), and skill and technique acquisition (1.00); the HLQ subscales of having sufficient information to manage my health (1.21) and actively managing my health (0.66); and the LINQ (1.85). Self-management, including activation, improved significantly in subjects with asthma or COPD who took part in a multidisciplinary pulmonary rehabilitation program.


Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders.
There are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5mM or sucrose 25 mM, and control (without additives) was cooled to 4 °C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P>0.05). However, EG3+S yielded the greatest percentages of the total abnormality (P<0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P<0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P<0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages of the tail intensity and tail moment (P<0.05). There were no significant differences observed in non-return rates among all treatment groups (P>0.05).


Post-infarction cardiac rupture.
Three allied conditions are described in this paper: (i) haemopericardium with cardiac rupture (5 cases); (ii) haemopericardium without rupture (2 cases); (iii) pseudoaneurysm (1 case). In the first 2 of these, the significant features were clinical deterioration with shock 3 or more days after infarction, recurrent cardiac pain, cardiac tamponade, and immediate or later ineffectiveness of counterpulsation. An additional feature in the second group was the development of haemopericardium after heparin therapy. In the third group, infarction followed by left ventricular failure and progressive cardiac enlargement was the significant feature. An apical systolic murmur was not present, as a false sac had not been formed. Ante-mortem diagnosis depends upon an appreciation of these features. Without it successful surgery is impossible. There were 4 survivors in this group of 8 patients.


Daily lifestyle behaviors and risks of sarcopenia among older adults.
Lifestyle behaviors are modifiable factors that can provide information for designing intervention strategies for sarcopenia. The present study aimed to identify the relationships between a range of daily lifestyle behaviors and sarcopenia risks among older adults. A nationwide telephone-based survey targeting older adults (≥65 years) was performed in Taiwan. Data based on self-reported daily lifestyle behaviors (food selection, physical activity, sitting time, and sleep duration), the presence or absence of sarcopenia (measured by SARC-F), and personal characteristics were obtained. Binary logistic regression models were applied. A total of 1068 older adults participated in this survey. In the adjusted model, older adults who selected unbalanced foods (odds ratio [OR] = 1.93, 95% confidence interval [CI] = 1.12-3.34), engaged in insufficient physical activity (OR = 5.14, 95% CI = 3.04-8.70), and sat for longer periods of time (OR = 1.98, 95% CI = 1.09-3.59) were more likely to have higher risks of sarcopenia. No significant association was observed for sleep duration. The results of this study highlight that, among health behaviors, an unbalanced food selection (six nutrients), not meeting physical activity recommendations (150 min/week), and a higher sitting time (≥7 h/day) were risk factors for sarcopenia among older adults. Intervention programs for sarcopenia prevention in older adults should focus on promoting balanced food selection, sufficient physical activity, and reduced sitting time.


Prospective study of restless legs syndrome and mortality among men.
To prospectively examine whether men with restless legs syndrome (RLS) had an increased risk of mortality. This was a prospective cohort study of 18,425 US men free of diabetes, arthritis, and renal failure in the Health Professionals Follow-up Study (HPFS). In 2002, RLS was assessed using a set of standardized questions. Deaths were identified from state vital statistics records, the National Death Index, family reports, and the postal system. During 8 years of follow-up (2002-2010), we documented 2,765 deaths. In an age-adjusted model, RLS was associated with a 39% increased risk of mortality (hazard ratio [HR] = 1.39; 95% confidence interval [CI] 1.19-1.62; p < 0.0001). The association between RLS and mortality was slightly attenuated after further adjustment for body mass index, lifestyle factors, chronic conditions, sleep duration, and other sleep-related disorders (adjusted HR = 1.30; 95% CI 1.11-1.52; p = 0.003). When we further excluded those with major chronic conditions (e.g., cancer, high blood pressure, cardiovascular disease, and other comorbidities), the adjusted HR was 1.92 (95% CI 1.03-3.56; p = 0.04). The interactions between RLS and other risk factors (older age, overweight, short sleep duration, smoking, low physical activity, and unhealthy diet) in relation to total mortality risk were not significant (p for interaction >0.2 for all). We observed that men with RLS had a higher overall mortality and this association was independent of known risk factors. The increased mortality in RLS was more frequently associated with respiratory disease, endocrine disease, nutritional/metabolic disease, and immunologic disorders. Future research exploring the pathophysiologic relationship between these disorders and RLS is warranted.


Persistent sexual, emotional, and cognitive impairment post-finasteride: a survey of men reporting symptoms.
Finasteride is a synthetic 5-α reductase inhibitor, which prevents the conversion of testosterone to dihydrotestosterone and has been used for more than 20 years in the treatment of male pattern hair loss. Randomized, controlled trials have associated finasteride with both reversible and persistent adverse effects. In this pilot study, we sought to characterize sexual and nonsexual adverse effects that men reported experiencing at least 3 months after stopping the medication. Based on previous research on persistent side effects of finasteride, we constructed an Internet survey targeting six domains: physical symptoms, sexual libido, ejaculatory disorders, disorders of the penis and testes, cognitive symptoms, and psychological symptoms and was e-mailed to patients who reported experiencing symptoms of side effects of finasteride. Responses from 131 generally healthy men (mean age, 24 years) who had taken finasteride for male pattern hair loss was included in the analysis. The most notable finding was that adverse effects persisted in each of the domains, indicating the possible presence of a "post-finasteride syndrome."


KNOX Protein OSH15 Induces Grain Shattering by Repressing Lignin Biosynthesis Genes.
Seed shattering is an agronomically important trait. Two major domestication factors are responsible for this: qSH1 and SH5. Whereas qSH1 functions in cell differentiation in the abscission zone (AZ), a major role of SH5 is the repression of lignin deposition. We have determined that a KNOX protein, OSH15, also controls seed shattering. Knockdown mutations of OSH15 showed reduced seed-shattering phenotypes. Coimmunoprecipitation experiments revealed that OSH15 interacts with SH5 and qSH1, two proteins in the BELL homeobox family. In transgenic plants carrying the OSH15 promoter-GUS reporter construct, the reporter gene was preferentially expressed in the AZ during young spikelet development. The RNA in situ hybridization experiment also showed that OSH15 messenger RNAs were abundant in the AZ during spikelet development. Analyses of osh15 SH5-D double mutants showed that SH5 could not increase the degree of seed shattering when OSH15 was absent, indicating that SH5 functions together with OSH15. In addition to the seed-shattering phenotype, osh15 mutants displayed dwarfism and accumulated a higher amount of lignin in internodes due to increased expression of the genes involved in lignin biosynthesis. Knockout mutations of CAD2, which encodes an enzyme for the last step in the monolignol biosynthesis pathway, caused an easy seed-shattering phenotype by reducing lignin deposition in the AZ This indicated that the lignin level is an important determinant of seed shattering in rice (Oryza sativa). Chromatin immunoprecipitation assays demonstrated that both OSH15 and SH5 interact directly with CAD2 chromatin. We conclude that OSH15 and SH5 form a dimer that enhances seed shattering by directly inhibiting lignin biosynthesis genes.


Statistics on cancer in China: cancer registration in 2002.
The objective of this study was to determine how many population-based cancer registries exist in China, what methods are being used, and the statistical data that are available from them, and to identify future needs with respect to technical support. A two-stage survey was conducted in 2002 at provincial and cancer registry level. Based on the questionnaire used in these two stages, the basic distribution and descriptive information on population-based cancer registry practices in China are addressed. There are 48 cancer registries in China, covering 73 million people (5.7% of the total population of China in 2000). The oldest three registries are LinZhou, ShangHai and QiDong. There are marked variations in practice between registries, with respect to data collection, data management and coding. Differences are also found in administrative aspects and sources of financial support. In conclusion, this first national survey of Chinese cancer registry practice provides a benchmark against which development and standardization can be evaluated in the future. The survey suggests that lack of qualified personnel, insufficient funding support and lack of stability of the population are major problems in carrying out registration work in China. It also indicates several ways in which registry practice, and hence availability and quality of incidence and survival data can be improved.


Neonatal lupus erythematosus: a report of four cases.
Neonatal lupus erythematosus is a very rare disease, clinically characterized by skin lesions that resemble those of subacute or discoid lupus erythematosus and/or congenital heart block. Generally, when patients have skin manifestations, they have no cardiac defects and vice-versa; however, in 10% of cases these manifestations may coexist. Other findings may include hematologic, hepatic and neurological abnormalities. This condition is caused by the transplacental passage of maternal autoantibodies against Ro (95%), La and, less frequently, U1-ribonucleoprotein (U1-RNP). The present case report describes four patients with clinical, histopathological and immunological findings compatible with neonatal lupus erythematosus, their treatment and progress.


Exposure-dose-response of Tellina deltoidalis to metal contaminated estuarine sediments 2. Lead spiked sediments.
Lead accumulation in estuarine sediments, as a result of activities such as mining and ore smelting, and through urban runoff is a continuing problem in the increasingly developed world. Marine organisms accumulate lead, which is known to be highly toxic to biological processes and to degrade organism and ecosystem health. Here the relationship between lead exposure, dose and response was investigated in the sediment dwelling, deposit feeding, marine bivalve Tellina deltoidalis. Bivalves were exposed in the laboratory to individual lead spiked sediments at < 0.01, 100 and 300 μg/g dry mass, for 28 days and accumulated total tissue lead concentrations of 4, 96 and 430 μg/g, respectively. Subcellular fractionation indicated that around 70% of the total accumulated tissue lead was detoxified, three quarters of the detoxified lead fraction was converted into metal rich granules, with the remainder in the metallothionein like protein fraction. The majority of biologically active lead was associated with the mitochondrial fraction with up to a 128 fold increase in lead burden in exposed organisms compared to controls. This indicates lead detoxification was occurring but the organism was unable to prevent lead interacting with sensitive organelles. With increased lead exposure T. deltoidalis showed a suppression in glutathione peroxidase activity, total glutathione concentration and reduced GSH:GSSG ratios, however, these differences were not significant. Lead exposed T. deltoidalis had a significantly reduced total antioxidant capacity which corresponded with increased lipid peroxidation, lysosomal destabilisation and micronuclei frequency. The exposure-dose-response relationships demonstrated for lead exposed T. deltoidalis supports its potential for the development of sublethal endpoints in lead toxicity assessment.


[TSH-secreting pituitary tumors: two case reports and literature review].
TSH-secreting pituitary adenomas are rare pituitary functioning tumors accounting for less than 2% of the pituitary adenomas. The clinical feature consists of thyrotoxicosis occasionally associated to tumoral symptoms due to mass effect. The biochemical feature consists of elevated thyroid hormones levels and normal or high TSH concentrations. This disease is often wrongly diagnosed as Grave's disease, and the ablative therapy is frequently conducted prior to the diagnosis. To report two cases followed in the Neuroendocrine Unit of Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo and to review the literature aiming at the management of this affection. In the presence of elevated thyroid hormone levels associated with inappropriate normal or increased TSH levels, the possibility of a TSH-secreting pituitary adenoma should be considered for the proper medical treatment.


Microparticulates: isolation from water and identification of associated chlorinated pesticides.
Microparticulates suspended in lake water were collected by continuous centrifugation and either examined directly or placed on a linear sucrose gradient. Total residue as well as fractions of the centrifuged gradient were extracted with hexane and examined by gas chromatography for the presence of chlorinated hydrocarbon pesticides. Hexane extracts of total residues were also examined by thin-layer chromatography. Lindane and endrin were shown, by gas-liquid chromatography and thin-layer chromatography, to be associated with microparticles. These and other pesticides appeared to be selectively associated with microparticles of different densities, when gas-liquid chromatography was used, although concentrations were below the detection limits required for confirmation by thin-layer chromatography. Samples taken at different times from different locations in Lake Erie revealed different associations with hexane-soluble electron-capturing compounds.


Disseminated Gonococcal Infection: A Case Report of Arthritis-Dermatitis Syndrome.
The bacteremic spread of the sexually transmitted pathogen, Neisseria gonorrhoeae (N. gonorrhoeae), results in disseminated gonococcal infection (DGI), which can lead to a variety of clinical signs and symptoms, such as multiple skin lesions, tenosynovitis and arthralgias/arthritis. The Centers for Disease Control and Prevention listed that drug-resistant N. gonorrhoeae (cephalosporin resistance) poses an urgent threat due to antimicrobial resistance. The authors describe the case of a young woman presenting in the emergency department (ED) with skin lesions and malaise. DGI was confirmed by blood cultures, nucleic acid amplification testing (NAAT) in cervical mucosa and urine gonococcal probe. One week of intravenous ceftriaxone was administrated with complete clinical and analytical resolution. The patient made a full clinical recovery. Clinicians must be aware of the signs and symptoms of this rare yet reemerging disease.


Cardiac transplantation for hypertrophic cardiomyopathy in the United States 2003-2011.
Cardiac transplant (CT) is the sole option in a minority of hypertrophic cardiomyopathy (HC) adults with refractory symptoms or end-stage disease. We aimed to examine the trends and hospital outcomes of CT in HC using 2003-2011 Nationwide Inpatient Sample database. HC comprised 1.1% of CT (151 of 14,277) performed during this time period (age 45±12 years, 67% male, 79% Caucasians). Number of HC CT increased from 2003 to 2011 (odds ratio=1.174; 95% confidence interval=1.102-1.252; P< 0.001). Comorbidities included congestive heart failure (76%), hypertension (23%), chronic kidney disease (23%), hyperlipidemia (19%), diabetes (13%), and coronary artery disease (10%). Acute in-hospital major adverse events occurred in 1 in 4 (23%) patient and 1 in 25 (3.8%) patients died perioperatively. Other major adverse events included allograft rejection or vasculopathy (23%), postoperative stroke or transient ischemic attack (3.5%), acute renal failure (43%), respiratory failure requiring mechanical ventilation (13%), sepsis (10%) or need for blood transfusion (10%). Compared to 1990-2004 United Network of Organ Sharing registry data (n=303), patients in current cohort had more comorbid conditions [diabetes (13%-vs-0%); chronic obstructive lung disease (9%-vs-1%); P < 0.001 for both), were more likely to be male (66%-vs-48% P< 0.001), were less likely to be Caucasian (79%-vs-86%; P < 0.001) or smokers (3%-vs-17%; P < 0.001) and less often required perioperative circulatory support or hemodialysis (17%-vs-49%, P < 0.001 and 3.2%-vs-8.3%, P = 0.04, respectively). HC comprises a small proportion of patients undergoing CT. The annual number of CT in HC has increased in recent years at least in part due to inclusion of patients with more comorbid conditions. Transplant recipients in the current cohort, however, required less postoperative circulatory support or renal replacement therapy.


Global relevance of Aire binding to hypomethylated lysine-4 of histone-3.
Aire promotes the ectopic expression of a repertoire of peripheral-tissue antigens (PTAs) in thymic medullary epithelial cells (MECs) to mediate deletional tolerance and thereby prevent autoimmunity. Binding of hypomethylated histone 3 (H3)-tails by Aire's plant homeodomain (PHD) finger is essential for Aire function in cultured cell models, prompting speculation that Aire-PHD:H3-tail interactions underlie targeting of Aire to weakly transcribed loci. To evaluate the role of Aire's PHD finger in MECs on a global scale in vivo, we complemented Aire-deficient mice with a mutant of Aire that inhibits its binding to hypomethylated H3K4 residues. Although the range of Aire-targeted genes was largely unaffected in these mice, the D299A mutation caused a global dampening of Aire's transcriptional impact, resulting in an autoimmune disease similar in profile to that of their Aire-deficient counterparts. To test whether a low H3K4 methylation state is sufficient for Aire targeting, we overexpressed an H3K4-specific demethylase in an Aire-dependent cultured cell system, and determined its capacity to extend Aire's transcriptional footprint. The range and magnitude of Aire-regulated genes was largely unaffected, the only genes additionally induced by Aire in this context being those already accessed for repression. In short, Aire's H3-binding module is necessary for Aire-mediated regulation of gene expression and central tolerance induction, but this influence is unlikely to reflect a targeting mechanism solely based on the recognition of hypomethylated H3K4 residues.


Occipital network for figure/ground organization.
To study the cortical mechanism of figure/ground categorization in the human brain, we employed fMRI and the temporal-asynchrony paradigm. This paradigm is able to eliminate any differential activation for local stimulus features, and thus to identify only global perceptual interactions. Strong segmentation of the image into different spatial configurations was generated solely from temporal asynchronies between zones of homogeneous dynamic noise. The figure/ground configuration was a single geometric figure enclosed in a larger surround region. In a control condition, the figure/ground organization was eliminated by segmenting the noise field into many identical temporal-asynchrony stripes. The manipulation of the type of perceptual organization triggered dramatic reorganization in the cortical activation pattern. The figure/ground configuration generated suppression of the ground representation (limited to early retinotopic visual cortex, V1 and V2) and strong activation in the motion complex hMT+/V5+; conversely, both responses were abolished when the figure/ground organization was eliminated. These results suggest that figure/ground processing is mediated by top-down suppression of the ground representation in the earliest visual areas V1/V2 through a signal arising in the motion complex. We propose a model of a recurrent cortical architecture incorporating suppressive feedback that operates in a topographic manner, forming a figure/ground categorization network distinct from that for "pure" scene segmentation and thus underlying the perceptual organization of dynamic scenes into cognitively relevant components.


Staged Surgery for Advanced Thyroid Cancers: Safety and Oncologic Outcomes of Neural Monitored Surgery.
Objective Thyroidectomy with extensive multicompartment bilateral neck dissections for advanced-stage thyroid cancer may lead to increased risk of complications, including bilateral recurrent laryngeal nerve (RLN) paralysis and hypoparathyroidism. A planned staged approach derived from a detailed preoperative radiographic map is associated with a low complication profile. This study evaluates oncologic results and safety of neural monitored, staged thyroid cancer surgery for management of advanced thyroid cancer. Study Design Case series with chart review. Setting Tertiary care center. Subjects and Methods With institutional review board approval, 35 consecutive patients with advanced thyroid malignancy and extensive nodal disease managed with staged surgery between January 2004 and May 2013 by the senior author (G.W.R.) were identified, and the oncologic and surgical outcomes were reviewed. Results In total, 37.2% of patients had stage III or IV disease, with extrathyroidal extension in 71.4%, vascular invasion in 51.4%, and RLN invasion in 17% of patients. A total of 34% patients had positive lymph nodes in more than 5 nodal compartments; the average positive lymph node yield was 17, and extranodal extension was present in 51%. Three patients had RLN sacrifice, and there were no other cases of temporary or permanent RLN paralysis; permanent hypoparathyroidism and chyle leak occurred in one patient each. Locoregional recurrence occurred in 5.7% of patients after a 147-week mean follow-up. In patients with papillary thyroid carcinoma, median postoperative nonstimulated and stimulated thyroglobulin levels were 0.2 and 0.75 ng/mL, respectively. Conclusion A neural monitored, staged surgical approach was conducted without significant adverse events in this small sample and represents and effective alternative strategy option to simultaneous bilateral surgery in the management of thyroid cancer with extensive neck metastases.


[Toxoplasmosis of the central nervous system in a diabetic patient].
The author describes a case of a central nervous system toxoplasmosis in a diabetic patient. A review of the literature about toxoplasmosis and its predisposing causes was made, and diabetes has not been reported up till now among them.


[The influence of the surgical and medicamental treatment on the olfactory function in the patients presenting with polypous rhinosinusitis].
The objective of the present investigation was to study dynamics of the olfactory function in the patients presenting with polypous rhinosinusitis receiving medicamental therapy and in different periods after the surgical intervention. A total of 40 patients were available for the examination in the "Sniffin Sticks" test. The patients of group 1 (n=20) were treated with intramuscular injections of dexamethasone (8 mg twice daily) with gradual tapering the dose during 10 days. The patients comprising group 2 underwent bilateral endoscopic polysinusotomy. Both treatment modalities were followed by momethasone fuorate therapy 6 months in duration. Examination of the patients on the first control visit (i.e. within 1 month after the onset of the treatment) revealed the significant improvement of all measured olfactory parameters that were not significantly different between the two groups. On the second control visit (within 4 months after the onset of the treatment), the patients of group 1 showed a significantly impaired ability to identify smells although other olfactory parameters remained virtually unaltered as compared with those on the first visit. At the same time, no significant impairment of the olfactory function was apparent in the patients of group 2. It is concluded that the patients having non-obstructive polyps in the nasal cavity should be advised in the first place to undergo a course of glucocorticosteroid therapy rather than surgical treatment because the results of conservative therapy and endoscopic polysinusotomy do not differ significantly, at least within the first 4 months after the beginning of the treatment.


Elemental maps from EFTEM images using two different background subtraction models.
Acquisition of a great number of energy-filtered images in a TEM (EFTEM) around the characteristic signal with a low energy-selecting slit allows display of the electron energy loss (EEL)-spectrum of regions of interest (ROIs) of a sample. These EEL-spectra can be submitted to the different treatments already in use for electron energy loss spectroscopy (EELS). In particular, it is possible to fit the experimental background with different mathematical models, using images acquired below and above a characteristic ionization edge. After this fitting, elemental maps can be computed by subtraction of the extrapolated/interpolated background from the characteristic images. In this work, we compared two mathematical models for background fitting-the Egerton power law and the log-polynomial law. We studied the low-energy region (40-150 eV) and a higher-energy region (350-600 eV) with the aid of software for interactive processing of EFTEM image series that we developed. The analyzed elements were the constitutive elements: iron, phosphorus, nitrogen, and oxygen in several biological materials. Two analytical TEMs, one equipped with a post-column and the other with an in-column spectrometer, were used. Our experimental results confirm that the power law is very sensitive to the value of the energy loss of the pre-edge images when the background is computed by extrapolation. The log-polynomial model is less sensitive than the power law model to the value of the energy loss of the pre-edge images in the low energy region. For the oxygen K edge at 535 eV, it gives the best fit when it is combined with the interpolation method. The use of programs that facilitate the handling of EFTEM image series, and the controlled calculation of the background under the characteristic images, represent a step forward in the generation of elemental maps.


Characteristics of Patients Involved in Physical Assault in an Acute Inpatient Psychiatric Setting.
Although aggressive behavior in psychiatric settings is a major concern, very few studies have focused exclusively on physical assault in a general inpatient psychiatric population. This study had 3 main goals: (1) to evaluate the prevalence of assaultive behavior in an acute psychiatric hospital; (2) to identify the clinical and socio-demographic factors associated with assaultive behavior during hospitalization; and (3) to explore whether a diagnosis of schizophrenia spectrum disorder increases the risk of assaultive behavior. We conducted a retrospective chart review of patients admitted to acute units in a psychiatric hospital between 2009 and 2012. A subset of occurrence reports identified by a multidisciplinary team as "physical assault" was included in the analysis. Using logistic multivariate regression analysis, these patients were compared with a randomly selected nonassaultive control group, matched for length of stay to identify factors associated with assaultive behavior. Of 757 occurrence reports, 613 met criteria for significant assault committed by 356 patients over 309,552 patient days. The assault incident density was 1.98 per 1000 patient days. In the logistic regression model of best fit, the factors significantly associated with assaultive behavior were age, legal status, and substance use. A diagnosis of schizophrenia spectrum disorder was not significantly associated with assaultive behavior. Clinicians should take extra precautions for involuntarily admitted young patients with a history of substance use, as they are more likely to exhibit assaultive behavior. A diagnosis of schizophrenia spectrum disorder in itself is not significantly associated with assaultive behavior. Screening instruments such as the Dynamic Appraisal of Situational Aggression may be useful in assessing risk of assault.


Mitochondrial autophagy in cells with mtDNA mutations results from synergistic loss of transmembrane potential and mTORC1 inhibition.
Autophagy has emerged as a key cellular process for organellar quality control, yet this pathway apparently fails to eliminate mitochondria containing pathogenic mutations in mitochondrial DNA (mtDNA) in patients with a variety of human diseases. In order to explore how mtDNA-mediated mitochondrial dysfunction interacts with endogenous autophagic pathways, we examined autophagic status in a panel of human cytoplasmic hybrid (cybrid) cell lines carrying a variety of pathogenic mtDNA mutations. We found that both genetic- and chemically induced loss of mitochondrial transmembrane potential (Δψ(m)) caused recruitment of the pro-mitophagic factor Parkin to mitochondria. Strikingly, however, the loss of Δψ(m) alone was insufficient to prompt delivery of mitochondria to the autophagosome (mitophagy). We found that mitophagy could be induced following treatment with the mTORC1 inhibitor rapamycin in cybrids carrying either large-scale partial deletions of mtDNA or complete depletion of mtDNA. Further, we found that the level of endogenous Parkin is a crucial determinant of mitophagy. These results suggest a two-hit model, in which the synergistic induction of both (i) mitochondrial recruitment of Parkin following the loss of Δψ(m) and (ii) mTORC1-controlled general macroautophagy is required for mitophagy. It appears that mitophagy can be accomplished by the endogenous autophagic machinery, but requires the full engagement of both of these pathways.


Clinical research with topical microbicides as a potential HIV prevention method.
An overview of the different phases in microbicide research is given with special attention to nonoxynol-9, the only ingredient that has been tested in phase III trials so far. The methodological and ethical challenges in conducting phase III studies are described. These include study population, compliance data, standardization, therapy for HIV-positive women, and informed consent. Despite these challenges, phase III trials are feasible and should be initiated now.


11-beta-hydroxysteroid dehydrogenase type 1 (HSD11B1) gene expression in muscle is linked to reduced skeletal muscle index in sarcopenic patients.
Glucocorticoids play a significant role in metabolic processes and pathways that impact muscle size, mass, and function. The expression of 11-beta-hydroxysteroid dehydrogenase type 1 (HSD11B1) has been previously described as a major regulator of skeletal muscle function in glucocorticoid-induced muscle atrophy and aging humans. Our study aimed to investigate glucocorticoid metabolism, including the expression of HSD11B1 in skeletal muscle, in patients with sarcopenia. Muscle biopsies were taken from the vastus lateralis muscle of thirty-three patients over 60 years of age with hip fractures. Sarcopenia status was assessed according to the criteria of the European Working Group on Sarcopenia in Older People 2. Skeletal muscle mass was measured by bioelectrical impedance analysis. Cortisol and cortisone concentrations were measured in serum. Gene expression analysis of HSD11B1, NR3C1, FBXO32, and TRIM63 in muscle biopsies was performed. Serial cross sections of skeletal muscle were labeled with myosin heavy chain slow (fiber type-1) and fast (fiber type-2) antibodies. The study included 33 patients (21 women) with a mean age of 82.5 ± 6.3 years, 17 patients revealed sarcopenic (n = 16 non-sarcopenic). Serum cortisone concentrations were negatively correlated with muscle mass (ß = - 0.425; p = 0.034) and type-2 fiber diameter (ß = - 0.591; p = 0.003). Gene expression of HSD11B1 (ß = - 0.673; p = 0.008) showed a negative correlation with muscle mass in the sarcopenic group. A significant correlation was found for the non-sarcopenic group for NR3C1 (ß = 0.548; p = 0.028) and muscle mass. These findings suggest a pathogenetic role of HSD11B1 in sarcopenic muscle.