Analysis of IL-1β, TGF-β, IL-5, ACE, PTPN22 gene polymorphisms, and gene expression levels in Turkish children with IgA vasculitis.
Immunoglobulin-A vasculitis (IgAV) is an inflammatory disease that affects small blood vessels. This study was performed to identify an association between protein tyrosine phosphatase non-receptor type 22 (PTPN22) + 788G > A (rs33996649), transforming growth factor-beta (TGF-β) -509C > T (rs18004069), interleukin 1-beta (IL-1β) -511C > T (rs16944), interleukin 5 (IL-5) -746C/T (rs2069812), and angiotensin-converting enzyme (ACE) I/D (rs4646994) gene polymorphisms, susceptibility to IgAV, as well as the mRNA levels of IL-1β, IL-1β, and TGF-β. A total of 53 patients with IgAV and 50 healthy controls were enrolled. PTPN22, TGF-β, IL-1β, ACE gene polymorphisms, ACE gene I/D polymorphisms, and mRNA expression levels were analyzed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, allele-specific PCR, and real-time PCR with TaqMan kits, respectively. PTPN22, TGF-β, IL-1β, IL-5, and ACE variants showed no genotype or allele differences between patients with IgAV and controls. Increased levels of IL-1β and TGF-β mRNA expressions were observed in patients with IgAV (p < 0.001). Patients with the IL-1β AG genotype showed significantly increased amounts of arthritis than patients with non-AG (p = 0.004). Age at disease onset was found to be significantly different in patients with IgAV according to the presence of TGF-β TT genotype (p = 0.047). Polymorphisms in PTPN22, TGF-β, IL-5, IL-1β, and ACE genes are unlikely to confer susceptibility to IgAV. However, the presence of the AG genotype of IL-1β is associated with susceptibility to IgAV-related arthritis. This is the first study to report a significant increase in serum mRNA levels of IL-1β and TGF-β in IgAV patients, supporting a susceptibility to IgAV in childhood.


Phase I trial of taxol given as a 3-hour infusion every 21 days.
Taxol is a unique plant product that promotes in vitro assembly of microtubules. In a phase I trial, adults with advanced solid tumors were given taxol (formulated with cremophor EL and dehydrated alcohol) as a 3-hour iv infusion every 21 days. The total dose administered ranged from 15 to 230 mg/m2 in nine escalation steps. Leukopenia, thrombocytopenia, nausea and vomiting, alopecia, stomatitis, transient rashes, increases in serum triglyceride levels, and hypersensitivity reactions were observed. Hypersensitivity reactions characterized by acute dyspnea, generalized erythema, and hypotension immediately following the initiation of the taxol infusion occurred in three of five patients receiving greater than or equal to 190 mg/m2 (18% of patients overall). No antitumor activity was observed. Hypersensitivity reactions constituted a treatment-limiting toxicity for this preparation of taxol given on this schedule over the dosage range examined. With the severity and unpredictability of the hypersensitivity reactions, further usage of taxol is not indicated with this drug formulation on this administration schedule. Further studies are warranted to uncover ways to permit the safe administration of taxol.


Complement gene variants determine the risk of immunoglobulin-associated MPGN and C3 glomerulopathy and predict long-term renal outcome.
Membranoproliferative glomerulonephritis (MPGN) is an uncommon cause of chronic nephropathy recently reclassified into immunoglobulin-associated MPGN (Ig-MPGN) and C3 glomerulopathy (C3G). In this study we aimed: (1) to evaluate the complement genetic and biochemical profile in patients with Ig-MPGN/C3G; (2) to investigate whether genetic variants and different patterns of complement activation (i.e., fluid versus solid phase) correlate with disease manifestations and outcomes. In 140 patients with idiopathic Ig-MPGN or C3G we performed complement biochemical and genetic screening and correlated genetic, biochemical and histology data with clinical features. Mutations in genes encoding alternative pathway complement proteins were found in both Ig-MPGN and C3G, and mutations in the two components of the C3 convertase are the most prevalent. We also report a mutation in THBD encoding thrombomodulin in a C3G patient. The presence of mutations alone does not significantly increase the risk of Ig-MPGN or C3G, but it does so when combined with common susceptibility variants (CD46 c.-366A in Ig-MPGN; CFH V62 and THBD A473 in C3G). Finally, patients without complement gene mutations or C3NeFs--autoantibodies that stabilize the alternative pathway C3 convertase--have a higher risk of progressing to end-stage renal disease than patients with identified mutations and/or C3NeFs, suggesting the existence of different pathogenetic mechanisms that lead to renal disease. We provide new insights into the pathogenesis of Ig-MPGN/C3G that underscore the complex nature of these diseases and suggest that the current C3G classification may miss many cases associated with abnormalities of the complement alternative pathway.


[Smoking and social inequality in Norway 1998-2000].
The Government wants a plan of action for equality in health, and smoking is the leading preventable cause of poor health and early death. Data were obtained from the national databank on smoking behaviour at the Directorate for Health and Social Affairs. Analyses to explore social gradients in terms of smoking status, intensity of consumption, product preference, age at first cigarette, and awareness of health hazards were run on responses from 5125 people above 24 from the years 1998-2002. There was a strong overrepresentation of smokers in lower socioeconomic groups. Smoking intensity is higher in these groups, and they tend also to smoke more dangerous products than smokers from higher socioeconomic groups. The age at first cigarette is lower on average, and they are more likely to condone passive smoking. Fewer take steps to limit smoking in the home and tend more frequently than others to be misinformed about the health hazards of different types of tobacco. The social gradient in current smoking behaviour poses a significant challenge to efforts to address health inequalities in the Norwegian population. Specifically tailored measures should be put in place for lower socioeconomic groups.


Pedigrees and Perpetrators: Uses of DNA and Genealogy in Forensic Investigations.
In the past few years, cases with DNA evidence that could not be solved with direct matches in DNA databases have benefited from comparing single-nucleotide polymorphism data with private and public genomic databases. Using a combination of genome comparisons and traditional genealogical research, investigators can triangulate distant relatives to the contributor of DNA data from a crime scene, ultimately identifying perpetrators of violent crimes. This approach has also been successful in identifying unknown deceased persons and perpetrators of lesser crimes. Such advances are bringing into focus ethical questions on how much access to DNA databases should be granted to law enforcement and how best to empower public genome contributors with control over their data. The necessary policies will take time to develop but can be informed by reflection on the familial searching policies developed for searches of the federal DNA database and considerations of the anonymity and privacy interests of civilians.


LncRNA FAM138B inhibits the progression of non-small cell lung cancer through miR-105-5p.
As a type of lung cancer, non-small cell lung cancer (NSCLC) has the characteristics of high mortality and high recurrence rate, which poses a great threat to human life and health. Due to the high risk of surgical treatment and the slow recovery of wounds, non-coding RNAs, especially lncRNAs are used as new potential clinical prognostic markers to prevent and treat cancer in advance. This study aims to explore the role of FAM138B in NSCLC and its possibility as a prognostic biomarker. Real-timequantitative polymerase chain reaction (RT-qPCR) was used to detect the expression and overexpression level of lncRNA FAM138B (FAM138B) in cells and tissues. The CCK-8, Transwell migration and invasion methods were performed to observe the cell transfection.The interaction between FAM138B and miR-105-5p was predicted by the bioinformatics tool starBase v2.0, and verified by the luciferase reporter gene experiment. Kaplan-Meier and Cox regression analyses were used to determine the prognostic significance of FAM138B in NSCLC. The expression of FAM138B is down-regulated in NSCLC cells and tissues. Overexpression of FAM138B can inhibit the expression level of miR-105-5p in NSCLC cells, and the ability of NSCLC cells to proliferate, migrate and invade is downregulated. FAM138B targets miR-105-5p, and there is a negative correlation between FAM138B and miR-105-5p. It is confirmed that FAM138B inhibits the progression of NSCLC by targeting miR-105-5p and can be a potential prognostic biomarker for NSCLC.


Whisking recovery after automated mechanical stimulation during facial nerve regeneration.
IMPORTANCE Recovery from facial nerve transection is typically poor, but daily mechanical stimulation of the face in rats has been reported to remarkably enhance functional recovery after facial nerve transection and suture repair. This phenomenon needs additional investigation because of its important clinical implications. OBJECTIVE To determine whether automated mechanical stimulation of the whisker pad improves whisking recovery after facial nerve transection and repair in a rat model. DESIGN AND SETTING Sixty-one rats underwent unilateral facial nerve transection and suture repair and were randomized into 8 groups. Six groups received daily automated whisker or whisker pad mechanical stimulation including 0.5-, 1.5-, and 8.0-Hz patterns. Two control groups received restraint without stimulation. Treatment started on postoperative day 8, occurred 5 days per week, and lasted throughout 15 weeks of recovery. Whisking amplitude, velocity, and acceleration were quantified weekly for 15 weeks. INTERVENTIONS Unilateral facial nerve transection, suture repair, and, for 6 groups, daily automated whisker or whisker pad mechanical stimulation. MAIN OUTCOMES AND MEASURES Quantification of whisking amplitude, velocity, and acceleration. RESULTS Rats receiving the low frequencies of stimulation of the whiskers or whisker pad did not demonstrate enhanced whisking recovery, and rats receiving stimulation at 8.0 Hz showed significantly worse whisking recovery compared with controls and previously published groups receiving lower dose manual stimulation. CONCLUSIONS AND RELEVANCE Although daily manual whisker pad stimulation has been shown to enhance whisking recovery, rats in this study did not demonstrate improved whisking recovery after automated mechanical stimulation across a wide range of driving frequencies. Moreover, faster stimulation (8.0 Hz) was actually detrimental to recovery. Further work is needed to understand the relationship between stimulation patterns and the physiologic mechanisms underlying improved or worsened functional outcomes after facial nerve transection and repair.


An international serotype 3 clone causing pediatric noninvasive infections in Israel, Costa Rica, and Lithuania.
Serotype 3 is known for its ability to cause invasive diseases worldwide. In the United States, after introduction of the 7-valent pneumococcal conjugate vaccine (PCV7), the prevalence of a serotype 3 clone (Netherlands-31/ST180) increased. The present study was aimed to evaluate the importance of serotype 3 clones in noninvasive infections in Israel, Costa Rica, and Lithuania. Molecular typing and antibiotic resistance were performed on 77 serotype 3 strains recovered from pediatric noninvasive infections during 2003-2005, and on 50 carried strains from healthy carriers. Serotype 3 ranked second among isolates from noninvasive infections in Costa Rica and Lithuania, and seventh among the Israeli isolates. Pulsed field gel electrophoresis (PFGE) analysis revealed the presence of 1 major cluster (64/77, 83%); this cluster comprised 60/64 fully susceptible strains that corresponded to the Netherlands-31/ST180 clone, and 4/64 multidrug-resistant strains, all from Lithuania, that corresponded to ST505, a double locus variant of ST180. Two additional fully susceptible clones, ST458 (11/77, 14%) and ST1116 (2/77, 3%), were found among the Israeli and Costa Rican strains, respectively. The same PFGE clusters identified among noninvasive infections were found among 50 isolates from carriers, with the same molecular characteristics. Serotype 3 accounts for a large proportion of mucosal disease in children, even before the introduction of PCV7. The data presented here describe for the first time the importance of a multidrug-resistant serotype 3 clone, ST505, in noninvasive infections.


[Burnout in intensive care units].
Intensive care units are highly stressful for the patients but for the caregivers as well, including nurse-assistants, nurses and physicians. The psychological syndrome of work exhaustion more commonly named burnout threatens these caregivers. The aims of the present paper are to describe: a) the incidence of burnout in intensive care units; b) the factors favoring burnout and c) the impacts of burnout at the individual, at the unit and institutional level. We suggest some possible ways to decrease the incidence of burnout. Finally, since the problematic of burnout is not specific to intensive care, we sought to underline some possible consequences of the burnout of caregivers on health systems.


Isolation and Functional Analysis of PISTILLATA Homolog From Magnolia wufengensis.
PISTILLATA (PI) homologs are crucial regulators of flower development in angiosperms. In this study, we isolated the MAwuPI homolog from Magnolia wufengensis, a basal angiosperm belonging to the Magnoliaceae. Molecular phylogenetic analysis suggested that MAwuPI was grouped into the PI/GLO lineages of B-class MADS-box gene with the distinctive PI motif. Further expression profiling analysis showed that MAwuPI was expressed in tepals and stamens but not in juvenile leaves and carpels, similar to the spatial expression pattern of AtPI in Arabidopsis. Interestingly, MAwuPI had higher expression level in inner-tepals than in outer-tepals, whereas the M. wufengensis flower is homochlamydeous. Moreover, ectopic expression of MAwuPI in Arabidopsis pi-1 mutant emerged filament-like structures but had no obvious petals, suggesting a partial phenotypic recovery of pi-1 mutant. The features of MAwuPI in the expression pattern and gene function improved our acknowledgment of B-class genes in M. wufengensis, and contributed to the clarification of M. wufengensis evolution status and relations with other sibling species in molecular perspective.


The role of positive and negative pressure on cavitation nucleation in nanodroplet-mediated histotripsy.
Nanodroplet-mediated histotripsy (NMH) is an ultrasound ablation technique combining histotripsy with acoustically sensitive perfluorocarbon (PFC) nanodroplets that can be selectively delivered to tumor cells for targeted tumor ablation. NMH takes advantage of the significantly reduced cavitation threshold of the nanodroplets, allowing for cavitation to be selectively generated only in regions containing nanodroplets. Understanding the physical mechanisms underlying the nanodroplet cavitation process is essential to the development of NMH. In this study, we hypothesize that cavitation nucleation is caused by the negative pressure (p-) exposed to the PFC, and the NMH cavitation threshold is therefore determined by the incident p- of the single-cycle pulses commonly used in NMH. This paper reports the first study that separately investigates the effects of negative and positive pressure on the NMH cavitation threshold using near half-cycle ultrasound pulses with dominant negative (negative-polarity pulses) or positive (positive-polarity pulses) pressure phases. Tissue phantoms containing perfluorohexane (PFH) nanodroplets were exposed to negative-polarity and positive-polarity pulses generated by a frequency compounding transducer recently developed in our lab, and the probability of generating cavitation was measured as a function of peak negative (p-) and peak positive (p+) pressure. The results showed close agreement in the p- cavitation threshold for PFH phantoms exposed to negative-polarity (11.4 ± 0.1 MPa) and positive-polarity (11.7 ± 0.2 MPa) pulses. The p+ at the cavitation threshold, in contrast, was measured to be sign ficantly different for the negative-polarity (4.0 ± 0.1 MPa) and positive-polarity (42.6 ± 0.2 MPa) pulses. In the final part of this study, the experimental results were compared to the cavitation threshold predicted by classical nucleation theory (CNT), with results showing close agreement between simulations and experiments. Overall, the results support our hypothesis and provide significant insight into the physical mechanisms underlying NMH.


Biallelic pathogenic variants of PARS2 cause developmental and epileptic encephalopathy with spike-and-wave activation in sleep.
Biallelic pathogenic variants in the mitochondrial prolyl-tRNA synthetase 2 gene (PARS2, OMIM * 612036) have been associated with Developmental and Epileptic Encephalopathy-75 (DEE-75, MIM #618437). This condition is typically characterized by early-onset refractory infantile spasms with hypsarrhythmia, intellectual disability, microcephaly, cerebral atrophy with hypomyelination, lactic acidemia, and cardiomyopathy. Most affected individuals do not survive beyond the age of 10 years. We describe a patient with early-onset DEE, consistently showing an EEG pattern of Spike-and-Wave Activation in Sleep (SWAS) since childhood. The patient underwent extensive clinical, metabolic and genetic investigations, including whole exome sequencing (WES). WES analysis identified compound heterozygous variants in PARS2 that have been already reported as pathogenic. A literature review of PARS2-associated DEE, focusing mainly on the electroclinical phenotype, did not reveal the association of SWAS with pathogenic variants in PARS2. Notably, unlike previously reported cases with the same genotype, this patient had longer survival without cardiac involvement or lactic acidosis, suggesting potential genetic modifiers contributing to disease variability. These findings widen the genetic heterogeneity of DEE-SWAS, including PARS2 as a causative gene in this syndromic entity, and highlight the importance of prolonged sleep EEG recording for the recognition of SWAS as a possible electroclinical evolution of PARS2-related DEE.


Effectiveness of bronchoscopic lung volume reduction using unilateral endobronchial valve: a systematic review and meta-analysis.
Bronchoscopic lung volume reduction (BLVR) can be suggested as an alternative for surgical lung volume reduction surgery for severe emphysema patients. This article intends to evaluate by systematic review the safety and effectiveness of BLVR using a one-way endobronchial valve. A systematic search of electronic databases, including MEDLINE, EMBASE, and the Cochrane Library, as well as eight domestic databases up to December 2013, was performed. Two reviewers independently screened all references according to selection criteria. The Scottish Intercollegiate Guidelines Network criterion was used to assess quality of literature. Data from randomized controlled trials were combined and meta-analysis was performed. This review included 15 studies. Forced expiratory volume in 1 second (FEV1) improved in the intervention group compared with the control group (mean difference [MD]=6.71, 95% confidence interval [CI]: 3.31-10.11). Six-minute walking distance (MD=15.66, 95% CI: 1.69-29.64) and cycle workload (MD=4.43, 95% CI: 1.80-7.07) also improved. In addition, St George's Respiratory Questionnaire score decreased (MD=4.29, 95% CI: -6.87 to -1.71) in the intervention group. In a subgroup analysis of patients with complete fissure, the FEV1 change from baseline was higher in the BLVR group than in the control group for both 6 months (MD=15.28, P<0.001) and 12 months (MD=17.65, P<0.001), whereas for patients with incomplete fissure, FEV1 and 6-minute walking distance showed no change. One-year follow-up randomized controlled trials reported deaths, although the cause of death was not related to BLVR. Respiratory failure and pneumothorax incidence rates were relatively higher in the BLVR group, but the difference was not significant. BLVR may be an effective and safe procedure for the treatment of severe COPD patients with emphysema, based on existing studies.


Oxidation by neutrophils-derived HOCl increases immunogenicity of proteins by converting them into ligands of several endocytic receptors involved in antigen uptake by dendritic cells and macrophages.
The initiation of adaptive immune responses to protein antigens has to be preceded by their uptake by antigen presenting cells and intracellular proteolytic processing. Paradoxically, endocytic receptors involved in antigen uptake do not bind the majority of proteins, which may be the main reason why purified proteins stimulate at most weak immune responses. A shared feature of different types of adjuvants, capable of boosting immunogenicity of protein vaccines, is their ability to induce acute inflammation, characterized by early influx of activated neutrophils. Neutrophils are also rapidly recruited to sites of tissue injury or infection. These cells are the source of potent oxidants, including hypochlorous acid (HOCl), causing oxidation of proteins present in inflammatory foci. We demonstrate that oxidation of proteins by endogenous, neutrophils-derived HOCl increases their immunogenicity. Upon oxidation, different, randomly chosen simple proteins (yeast alcohol dehydrogenase, human and bovine serum albumin) and glycoproteins (human apo-transferrin, ovalbumin) gain the ability to bind with high affinity to several endocytic receptors on antigen presenting cells, which seems to be the major mechanism of their increased immunogenicity. The mannose receptor (CD206), scavenger receptors A (CD204) and CD36 were responsible for the uptake and presentation of HOCl-modified proteins by murine dendritic cells and macrophages. Other scavenger receptors, SREC-I and LOX-1, as well as RAGE were also able to bind HOCl-modified proteins, but they did not contribute significantly to these ligands uptake by dendritic cells because they were either not expressed or exhibited preference for more heavily oxidised proteins. Our results indicate that oxidation by neutrophils-derived HOCl may be a physiological mechanism of conferring immunogenicity on proteins which in their native forms do not bind to endocytic receptors. This mechanism might enable the immune system to detect infections caused by pathogens not recognized by pattern recognition receptors.


Disparities in Medical Assessment Practices for Adolescents at Risk for Eating Disorders.
The United States Preventative Services Task Force found insufficient evidence to support universal screening for eating disorders (EDs) but did recommend assessing high-risk adolescents through laboratory tests, close follow-up, and referrals to other specialties. Yet, it is unclear whether youth at high risk for EDs receive such assessment and whether patient characteristics influence such practices. Using the Rochester Epidemiological Project, we identified adolescents (13-18 years) at risk for EDs (i.e., weight loss, underweight, or loss of appetite not explained by a medical condition) who presented for a medical appointment between January 1, 2005 to December 31, 2017 (n = 662; M age = 15.8 years; 66% female; 76% white). Patient and visit characteristics, assessment practices (i.e., tests, referrals, and follow-up), and ED diagnoses within 5 years following index visit were extracted. Adolescents who received referrals to other providers were 4 times more likely to be diagnosed with a future ED (p < .001) and were diagnosed 137.8 days sooner (Est = -137.8, p = .04) compared to those who did not receive referrals. Compared to males, females were 2.2 times more likely to receive referrals (p < .001). Compared to those presenting at a lower body mass index, adolescents with a higher body mass index were more likely to receive medical tests (HR = 1.0, p < .01) and less likely to receive recommendations to improve eating/weight (HR = 0.99, p < .01) or follow up visits (HR = 0.99, p < .01). Disparities in assessment practices for adolescents at high-risk for EDs underscore the need for improved tools to enhance early detection and treatment.


Nucleic acid synthesis in bacteriophage SPO2c 1 -infected Bacillus subtilis.
The synthesis of host macromolecules was shut off very slowly and incompletely by bacteriophage SPO2c(1). No change in the rate of incorporation of radioactive precursors into protein and ribonucleic acid (RNA) could be detected after infection, and the rate of incorporation of thymidine was increased only slightly. The relative proportions of phage and host species of nucleic acids at various intervals in the latent period were determined by means of nucleic acid hybridization. Phage-specific RNA populations synthesized early were different from those synthesized late in the latent period. Host deoxyribonucleic acid (DNA) replication continued until 8 to 10 min after SPO2c(1) infection and then decreased markedly as phage-specific DNA synthesis was initiated. Host DNA was not degraded to trichloroacetic acid-soluble fragments, and its nucleotides were not found in either newly synthesized intracellular phage DNA or in progeny phage particles. The average burst size of SPO2c(1) was approximately 200 plaque-forming units per cell.


[Development of Fluorescence Sensing Mechanism for Cell Functional Analysis].
Fluorescence probes are now widely used as indispensable tools in many cell functional analyses. At present, the design of fluorescent probes largely depends on the limited numbers of established sensing mechanisms such as photo-induced electron transfer (PET), photo-induced charge transfer (PCT), and fluorescence resonance energy transfer (FRET). Although these mechanisms are versatile in metal ion sensing, introduction of a new sensing mechanism is highly desirable not only to design a more sophisticated probe with high selectivity and sensitivity but also to expand the diversity of the sensing targets to unveil biological phenomena. In this article, we report the design of dual emission fluorescent probes for metal ions based on a unique fluorescence-sensing mechanism. The fluorescent probes incorporating this sensing mechanism displayed a large emission red-shift on complexation with metal ions such as Cd(II) and Ag(I). X-ray crystallography and theoretical computational calculations of the Cd(II) and Ag(I) complexes revealed that the emission shift was induced by non-coordination contact between the aromatic ring of fluorophore and the metal ion (arene-metal ion contact; AM-contact), which modulates the energy levels of molecular orbitals. The fluorescent probe was successfully applied to in cell ratiometric bioimaging of bioactive hydrogen sulfide (H2S). These successful applications highlight the usefulness of this sensing mechanism in biological fluorescence analysis.


Analysis of virus-specific mRNAs present in cells transformed with restriction fragments of adenovirus type 5 DNA.
Adenovirus type 5 (Ad5) mRNAs present in cells transformed with left-terminal Ad5 DNA fragments (XhoI-C, 0 to 15.5%; HindIII-G, 0 to 7.7%; HpaI-E, 0 to 4.3% were characterized by 'Northern blotting' and S1 nuclease analysis. They were compared with the mRNAs transcribed from the Ad5 E1 region in the early and late stages of lytic infection. It is shown that in XhoI-C-transformed cells the same mRNAs were transcribed as early during lytic infection: two co-terminal mRNAs from region E1a, differing only in their splicing, and one major E1b transcript. In HindIII-G-transformed cells additional E1a mRNAs were detected with a novel 5' terminus, but with the normal splicing pattern. Instead of the normal E1b mRNA, HindIII-G-transformed cells were found to contain mRNAs consisting of a viral E1b segment and a non-viral segment. This E1b-encoded segment was shown not to be involved in RNA splicing. The mRNAs in cells transformed with Ad5 HpaI-E were similar to the E1a mRNAs found in XhoI-C- and HindIII-G-transformed, and in lytically infected cells but had aberrant 3' termini. These results are discussed in the light of the Ad5 E1 DNA and RNA sequences, and protein mapping data.


Neurological Diseases With Autism Spectrum Disorder: Role of ASD Risk Genes.
Autism spectrum disorder (ASD) is frequently comorbid with other neurological disorders such as intellectual disability (ID) or global development delay (GDD) and epilepsy. The pathogenesis of ASD is complex. So far, studies have identified more than 1000 ASD risk genes. Most of them were also reported to relate with other neurological diseases, and only several of them have been confirmed as pathogenic genes for autism. Little is known about the roles of these risk genes in neurological diseases with ASD. In the present study, we recruited a cohort of 158 neurological disorder probands with 163 variants of 48 ASD risk genes. Of these, 50 individuals (31.6%) were diagnosed with ASD. In the ASD patient subset, we identified several rarely reported candidate genes including DOLK, USH2A, and HUWE1. In a comparison of patients with neurological disorders with and without ASD, we found that ID/GDD was frequently comorbid with ASD whereas epilepsy was more common in the non-ASD group. Statistical analyses of all possible risk factors implicated that variants in synaptic genes, especially non-voltage-gated ion channel genes and in transcriptional and chromosome genes were related to ASD, but none of the investigated environmental factors was. Our results are useful for the future diagnosis and prognosis of patients with neurological disorders and emphasize the utility of genetic screening.


Phase II trial of oral etoposide in recurrent or refractory endometrial adenocarcinoma: a southwest oncology group study.
Oral etoposide has activity in a wide variety of tumors and is well tolerated. Therefore, the efficacy of oral etoposide was assessed as a treatment of metastatic endometrial cancer. To be eligible for this group-wide Southwest Oncology Group trial, patients had to have histologically proven metastatic or recurrent endometrial carcinoma; no previous cytotoxic therapy; and adequate renal, hepatic, and hematologic function, and they had to have given informed consent. Therapy consisted of oral etoposide, 50 mg daily on days 1-21 on a 28-day schedule. Therapy was continued in the absence of toxicity or disease progression. Forty-four eligible women, with a median age of 68 years (range 38-84 years) were treated. Radiotherapy had been delivered to 33 and hormomal therapy to 21. The median duration of therapy was 69 days (range 7-510 days). The treatment was well tolerated. Only one patient had grade 4 neutropenia, and a second had grade 4 anemia. Three patients had grade 3 nausea. One complete and five partial responses (14%) were observed. An additional four patients had unconfirmed responses. Tumor regressions were noted in nodes, bone, and visceral organs. While oral etoposide has only a modest level of activity when used in chemonaive patients, the minimal toxicity of this drug makes it a candidate for use in combination chemotherapy.


Optimizing, validating, and field testing a multiplex qPCR for the detection of amphibian pathogens.
Amphibian populations worldwide are facing numerous threats, including the emergence and spread of infectious diseases. In the past 2 decades, Batrachochytrium dendrobatidis (Bd), a parasitic fungus, and a group of viruses comprising the genus Ranavirus have become widespread and resulted in mass mortality events and extirpations worldwide. In 2013, another novel fungus, B. salamandrivorans (Bsal), was attributed to dramatic declines in populations of fire salamander Salamandra salamandra in the Netherlands. Experimental infections demonstrated that Bsal is highly pathogenic to numerous salamander genera. In an effort to prevent the introduction of Bsal to North America, the US Fish and Wildlife Service (USFWS) listed 201 salamander species as injurious wildlife under the Lacey Act. To determine infection status and accurately assess amphibian health, the development of a sensitive and specific diagnostic assay was needed. We describe the optimization and validation of a multiplex quantitative polymerase chain reaction (qPCR) protocol for the simultaneous detection of Bd, Bsal, and frog virus 3-like ranaviruses. A synthetic genome template (gBlock®) containing the target genes from all 3 pathogens served as the positive control and allowed accurate quantification of pathogen genes. The assay was validated in the field using an established non-lethal swabbing technique to survey local amphibian populations throughout a range of habitats. This multiplex qPCR demonstrates high reproducibility, sensitivity, and was capable of detecting both Bd and ranavirus in numerous locations, species, and life stages. Bsal was not detected at any point during these sampling efforts.


Using candidacy theory to explore unemployed over-50s perceptions of suitability of a welfare to work programme: A longitudinal qualitative study.
Welfare to work interventions seek to move out-of-work individuals from claiming unemployment benefits towards paid work. However, previous research has highlighted that for over-50s, particularly those with chronic health conditions, participation in such activities are less likely to result in a return to work. Using longitudinal semi-structured interviews, we followed 26 over-50s during their experience of a mandated welfare to work intervention (the Work Programme) in the United Kingdom. Focusing on their perception of suitability, we utilise and adapt Candidacy Theory to explore how previous experiences of work, health, and interaction with staff (both in the intervention, and with healthcare practitioners) influence these perceptions. Despite many participants acknowledging the benefit of work, many described a pessimism regarding their own ability to return to work in the future, and therefore their lack of suitability for this intervention. This was particularly felt by those with chronic health conditions, who reflected on difficulties with managing their conditions (e.g., attending appointments, adhering to treatment regimens). By adapting Candidacy Theory, we highlighted the ways that mandatory intervention was navigated by all the participants, and how some discussed attempts to remove themselves from this intervention. We also discuss the role played by decision makers such as employment-support staff and healthcare practitioners in supporting or contesting these feelings. Findings suggest that greater effort is required by policy makers to understand the lived experience of chronic illness in terms of ability to RTW, and the importance of inter-agency work in shaping perceptions of those involved.


beta-endorphin suppresses adrenocorticotropin and cortisol levels in normal human subjects.
To determine the effect of beta-endorphin on the pituitary-adrenal axis, human synthetic beta-endorphin was infused iv in 10 normal human subjects. Either beta-endorphin (0.3, 1.0, and 3.0 micrograms/kg . min, each dose for 30 min) or a control (sham) infusion of 5% dextrose water was administered in a blind fashion in the same subjects on 2 successive days. Plasma ACTH and cortisol and serum human GH and PRL levels were measured 30 min before and then every 30 min for 210 min during and after both the beta-endorphin and control infusions. In all subjects, cortisol levels decreased below the basal level in response to the infusion of beta-endorphin. The threshold dose was 1.0 micrograms/kg . min, with the mean control cortisol level (12 +/- 2 micrograms/dl) declining significantly to 7 +/- 1 micrograms/dl (1.0 micrograms/kg . min) and 6 +/- 1 micrograms/dl (3.0 micrograms/kg . min; P less than 0.01). ACTH levels also were significantly lower than the control value (48 +/- 6 pg/ml) at the 1.0 microgram/kg . min dose (32 +/- 4 pg/ml; P less than 0.05). The decline in ACTH and cortisol levels was also significantly different from levels obtained during the control infusions (P = 0.001 and P = 0.01, respectively). The results are consistent with short feedback loop inhibition of pituitary ACTH release or suppression of hypothalamic corticotropin-releasing factor release by beta-endorphin.


The nature of pristine noble gases in mantle plumes
High-precision noble gas data show that the Hawaiian and Icelandic mantle plume sources contain uniquely primitive neon that is composed of moderately nucleogenic neon-21 and a primordial component indistinguishable from the meteoritic occurrence of solar neon. This suggests that Earth's solar-type rare gas inventory was acquired during accretion from small planetesimals previously irradiated by solar wind from the early sun. However, nonradiogenic argon, krypton, and xenon isotopes derived from the mantle display nonsolar compositions and indicate an atmosphere-like fingerprint that is not due to recent subduction.


Duration of apnoea with two different intubating doses of remifentanil.
We have studied the apnoea time after induction with two successful drug intubating regimes, both containing remifentanil. Group 1 (n = 20); propofol 2 mg kg-1 and remifentanil 2 micrograms kg-1 and group 2 (n = 20); propofol 2 mg kg-1, remifentanil 1 microgram kg-1 and lignocaine 1 mg kg-1. Intubation was possible in all 40 patients, and regarded as acceptable in 90% and 85% of patients, respectively. The median time to the return of spontaneous respiration was 487 s in group 1 and 270 s in group 2 (P < 0.05). Median end-tidal CO2 concentration at the time of the first spontaneous respiration was 7.37 kPa in group 1 and 6.3 kPa in group 2 (P < 0.05). Both groups had a decrease in heart rate after induction, but this failed to reach either clinical or statistical significance, and no patient required atropine. Similarly, there was a decrease in arterial pressure after induction in both groups (P < 0.05), but this was not deemed to be clinically significant. Only two patients in each group required ephedrine 6 mg before arterial pressure was restored to within 25% of the base-line. Intubating conditions were similar in both groups, but group 2 provided a significantly shorter apnoea time.


Role of methylation in the induced and spontaneous expression of the avian endogenous virus ev-1: DNA structure and gene products.
The endogenous avian provirus ev-1 is widespread in white leghorn chickens. Although it has no major structural defects, ev-1 has not been associated with any phenotype and is ordinarily expressed at a very low level. In this report, we describe a chicken embryo (Number 1836) cell culture containing both ev-1 and ev-6 which spontaneously expressed the ev-1 provirus. This culture released a high level of noninfectious virions containing a full complement of virion structural (gag) proteins but devoid of reverse transcriptase activity or antigen. These virions contained 70S RNA closely related to the genome of Rous-associated virus type 0, but identifiable as the ev-1 genome by oligonucleotide mapping. A fraction of the RNA molecules in the 70S complex were unusual in that they were polyadenylated 100 to 200 nucleotides downstream of the usual polyadenylation site. Eight sibling embryo cultures did not share this unusual phenotype with 1836, indicating that it was not inherited. However, an identical phenotype was inducible in the sibling cultures by treatment with 5-azacytidine, an inhibitor of DNA methylation, and the induced expression was stable for more than 10 generations. Analysis of chromatin structure and DNA methylation of the ev-1 provirus in 1836 cells revealed the presence (in a fraction of the proviruses) of both DNase I hypersensitive sites in the long terminal repeats and in gag and a pattern of cleavage sites for methyl-sensitive restriction endonuclease not found in a nonexpressing sibling. These results lend strong support to the role of DNA methylation in the control of gene expression. Additionally, they explain the lack of phenotype associated with ev-1 as due to a combination of its low expression and defectiveness in pol and env.


Chlamydophila psittaci eradication with doxycycline as first-line targeted therapy for ocular adnexae lymphoma: final results of an international phase II trial.
The pathogenic association between Chlamydophila psittaci (Cp) and ocular adnexal marginal zone lymphoma (OAMZL) and the efficacy of doxycycline monotherapy have been investigated in retrospective series with variations in stage, management, and follow-up duration. To our knowledge, this is the first international phase II trial aimed at clarifying Cp prevalence and activity of first-line doxycycline in a homogeneous series of consecutive patients with newly diagnosed stage I OAMZL. Forty-seven patients were registered. Tumor tissue, conjunctival swabs, and peripheral blood from 44 patients were assessed for seven Chlamydiaceae infections by three polymerase chain reaction protocols. Thirty-four patients with measurable or parametrable disease were treated with doxycycline and assessed for chlamydial eradication and lymphoma response (primary end point). Cp DNA was detected in biopsies of 39 patients (89%); no other Chlamydiaceae were detected. Twenty-nine patients had Cp DNA in baseline swabs and/or blood samples and were evaluable for chlamydial eradication, which was achieved in 14 patients (48%). Lymphoma regression was complete in six patients and partial in 16 (overall response rate, 65%; 95% CI, 49% to 81%); 11 had stable disease, and one had progressive disease. At a median follow-up of 37 months (range, 15 to 62 months), 20 patients remained relapse free (5-year progression-free survival [PFS] ± standard deviation, 55% ± 9%). Cp eradication was associated with improved response rate (86% v 47%; P = .02) and 5-year PFS (68% v 47%; P = .11). Upfront doxycycline is a rational and active treatment for patients with stage I Cp-positive OAMZL. Lymphoma regression is consequent to Cp eradication, which can easily be monitored on conjunctival and blood samples. Prospective trials aimed at identifying more effective administration schedules for doxycycline are warranted.


The influence of environmental context in interpersonal observation-execution.
Cyclical upper-limb movements involuntarily deviate from a primary movement direction when the actor concurrently observes incongruent biological motion. We examined whether environmental context influences such motor interference during interpersonal observation-execution. Participants executed continuous horizontal arm movements while observing congruent horizontal or incongruent curvilinear biological movements with or without the presence of an object positioned as an obstacle or distractor. When participants were observing a curvilinear movement, an object located within the movement space became an obstacle, and, thus, the curvilinear trajectory was essential to reach into horizontal space. When acting as a distractor, or with no object, the curvilinear trajectory was no longer essential. For observing horizontal movements, objects were located at the same relative locations as in the curvilinear movement condition. We found greater involuntary movement deviation when observing curvilinear than horizontal movements. Also, there was an influence of context only when observing horizontal movements, with greater deviation exhibited in the presence of a large obstacle. These findings suggest that the influence of environmental context is underpinned by the (mis-)matching of observed and executed actions as incongruent biological motion is primarily coded via bottom-up sensorimotor processes, whilst the congruent condition incorporates surrounding environmental features to modulate the bottom-up sensorimotor processes.


Immune thrombocytopenia with clinical significance in systemic lupus erythematosus: a retrospective cohort study of 90 patients.
To describe the characteristics, treatment and outcome of patients with immune thrombocytopenia with clinical significance (ITPCS) associated with SLE. This retrospective multicentre study included SLE patients who experienced ≥1 ITPCS (defined as ITP with attributable bleeding disorders and/or a platelet count <30×109/l). Other causes of secondary thrombocytopenia were excluded. Major bleeding event (MBG) was defined as Khellaf score >8 and/or WHO score >2. A total of 90 patients were included, the median (range) follow-up duration was 80 (6-446) months. ITP was diagnosed before SLE in 25 patients. They presented a high rate of autoimmune haemolytic anaemia (15%), antiphospholipid antibody (62%) and antiphospholipid syndrome (19%). The 25 (28%) patients who experienced MBG had significantly more bleedings at ITP diagnosis and higher bleeding scores, and serositis and thrombosis during follow-up. They required significantly more treatment lines, transfusions and hospitalizations. The 11 (12%) patients who experienced no bleeding event presented a significantly more restricted SLE phenotype (cutaneous and/or articular). Patients received a mean (range) of 4.2 (1-11) treatment lines. Corticosteroids and HCQ allowed ITPCS overall response in one-third of patients. The median relapse-free survival of rituximab (n = 34), AZA (n = 19), MMF (n = 8), thrombopoietin-receptor agonists (n = 16) and splenectomy (n = 19) were 53, 31.5, 61, 24.5 and 78 months, respectively. Four patients experienced thrombotic events after splenectomy and one occurred under thrombopoietin-receptor agonist treatment. SLE-ITCS patients displayed a high rate of haematological abnormalities and MBG patients exhibited higher morbidity. Management of thrombocytopenia was highly heterogeneous and many options seem viable.


Recent breakthroughs on the development of electrodeionization systems for toxic pollutants removal from water environment.
Since ecosystems are becoming inherently polluted, long-term contaminant removal methods are required. Electrodeionization, in particular, has recently been demonstrated as an effective approach for eliminating ionic compounds from contaminated water sources. Being a more environmentally friendly technology is most likely the main reason for its eminence. It uses electricity to replace toxic contaminants that are conventionally used to regenerate and hence reducing the toxins associated with resin regeneration. In wastewater treatment, continuous electrodeionization system overcomes several limitations of ion exchange resins, notably ion dumping. This prospective assessment delves into the mechanism, principle, and theory of electrodeionization system. It also focused on the design and applications, particularly in the removal of toxic compounds, as well as current advances in the electrodeionization system. Recent breakthroughs in electrodeionization were comprehensively discussed. Further developments in electrodeionization systems are also projected, with improved efficiency at the time of functioning at lower costs because of reduced energy use, proving them desirable for commercial usage with a broad array of applications across the globe.


Methylation of histone H3 lysine 36 is a barrier for therapeutic interventions of head and neck squamous cell carcinoma.
Approximately 20% of head and neck squamous cell carcinomas (HNSCC) exhibit reduced methylation on lysine 36 of histone H3 (H3K36me) due to mutations in histone methylase NSD1 or a lysine-to-methionine mutation in histone H3 (H3K36M). Whether such alterations of H3K36me can be exploited for therapeutic interventions is still unknown. Here, we show that HNSCC models expressing H3K36M can be divided into two groups: those that display aberrant accumulation of H3K27me3 and those that maintain steady levels of H3K27me3. The first group shows decreased proliferation, genome instability, and increased sensitivity to genotoxic agents, such as PARP1/2 inhibitors. In contrast, the H3K36M HNSCC models with steady H3K27me3 levels do not exhibit these characteristics unless H3K27me3 levels are elevated, either by DNA hypomethylating agents or by inhibiting the H3K27me3 demethylases KDM6A/B. Mechanistically, we found that H3K36M reduces H3K36me by directly impeding the activities of the histone methyltransferase NSD3 and the histone demethylase LSD2. Notably, we found that aberrant H3K27me3 levels induced by H3K36M expression is not a bona fide epigenetic mark in HNSCC since it requires continuous expression of H3K36M to be inherited. Moreover, increased sensitivity of H3K36M HNSCC models to PARP1/2 inhibitors solely depends on the increased H3K27me3 levels. Indeed, aberrantly high H3K27me3 levels decrease BRCA1 and FANCD2-dependent DNA repair, resulting in higher sensitivity to DNA breaks and replication stress. Finally, in support of our in vitro findings, a PARP1/2 inhibitor alone reduce tumor burden in a H3K36M HNSCC xenograft model with elevated H3K27me3, whereas in a H3K36M HNSCC xenograft model with consistent H3K27me3 levels, a combination of PARP1/2 inhibitors and agents that upregulate H3K27me3 proves to be successful. In conclusion, our findings underscore a delicate balance between H3K36 and H3K27 methylation, essential for maintaining genome stability. This equilibrium presents promising therapeutic opportunities for patients with H3K36me-deficient tumors.


The effects of hyperhomocysteinemia on the microcirculation of skin flaps.
Hyperhomocysteinemia is an independent risk factor for atherothrombotic disease and venous thrombosis. The effects of hyperhomocysteinemia on the microcirculation were studied in vascular diseases. The authors aimed to investigate the effects of hyperhomocysteinemia on the microcirculation of random-pattern skin flaps. Twenty-two male Sprague-Dawley rats, divided into two groups, were used in this study. The rats in group 1 (control) were fed the TD.07112 diet, and the rats in group 2 (experimental group) were fed the TD.07114 diet, enriched in methionine for 30 days, to induce severe hyperhomocysteinemia. The plasma homocysteine, folic acid, vitamin B12, and vitamin B6 levels were evaluated on days 0 and 30. Distally based skin flaps were elevated on day 30 and evaluated by direct observation, microangiography, and light microscopy on day 37. Mean homocysteine blood levels were 211.76 ± 56.55 μM/liter in group 2 and 14.48 ± 2.00 μM/liter in group 1 on day 30. The rate of necrosis was significantly higher in group 2 (59.00 ± 4.38 percent) compared with group 1 (32.54 ± 6.13 percent; p < 0.01). Microangiographic findings were similar to direct observation results. Microvessel calibration was reduced in group 2. In group 1, structures of epidermis and dermis were normal; however, there was a slight mononuclear cell infiltration along with thick collagen fibers. A more prominent mononuclear cell infiltration with fields of loose epidermis, associated with inflammation and infiltration, were observed in group 2. The authors demonstrated, for the first time, that hyperhomocysteinemia severely suppressed the microvasculature of skin flaps, as shown by increased flap necrosis and reduced microvessel calibration in the experimental group.


Vascular disease in COPD: Systemic and pulmonary expression of PARC (Pulmonary and Activation-Regulated Chemokine).
The role of Pulmonary and Activation-Regulated Chemokine (PARC) in the physiopathology of Chronic Obstructive Pulmonary Disease (COPD) is not fully understood. The aim of the present study is to analyze the expression of PARC in lung tissue and its relationship with the vascular remodeling of the systemic and pulmonary arteries of COPD subjects. To achieve this objective, protein and gene expression experiments, together with ELISA assays, were performed on the lung tissue, intercostal arteries and serum samples from COPD patients, non-obstructed smokers (NOS) and never-smokers (NS). A total of 57 subjects were included in the analysis (23 COPD, 18 NOS and 16 NS). In the comparisons between groups, a significantly increased lung protein expression of PARC was observed in the COPD group compared to the NOS group (1.96±0.22 vs. 1.29±0.27, P-adjusted = 0.038). PARC was located predominantly in the smooth muscle cells of the remodeled pulmonary muscular arteries and the macrophage-rich area of the alveolar parenchyma. No differences were detected in PARC gene expression analyses. The protein content of PARC in the intercostal arteries were similar between groups, though little remodeling was observed in these arteries. Circulating levels of PARC were numerically higher in patients with COPD compared to NOS and NS. The results of the present study suggest an increased lung protein expression of PARC in COPD subjects. This protein was mainly localized in the smooth muscle cells of the pulmonary muscular arteries and was associated with the severity of intimal thickening, indicating its possible role in this remodeling process.


Herbal Medicine in the Treatment of Non-Alcoholic Fatty Liver Diseases-Efficacy, Action Mechanism, and Clinical Application.
Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disease with high prevalence in the developed countries. NAFLD has been considered as one of the leading causes of cryptogenic cirrhosis and chronic liver disease. The individuals with obesity, insulin resistance and diabetes mellitus, hyperlipidaemia, and hypertension cardiovascular disease have a high risk to develop NAFLD. The related critical pathological events are associated with the development of NAFLD including insulin resistance, lipid metabolism dysfunction, oxidative stress, inflammation, apoptosis, and fibrosis. The development of NAFLD range from simple steatosis to non-alcoholic steatohepatitis (NASH). Hepatic steatosis is characterized by fat accumulation, which represents the early stage of NAFLD. Then, inflammation triggered by steatosis drives early NAFLD progression into NASH. Therefore, the amelioration of steatosis and inflammation is essential for NAFLD therapy. The herbal medicine have taken great effects on the improvement of steatosis and inflammation for treating NAFLD. It has been found out that these effects involved the multiple mechanisms underlying lipid metabolism and inflammation. In this review, we pay particular attention on herbal medicine treatment and make summary about the research of herbal medicine, including herb formula, herb extract and naturals compound on NAFLD. We make details about their protective effects, the mechanism of action involved in the amelioration steatosis and inflammation for NAFLD therapy as well as the clinical application.


HEAD AND THORAX ELEVATION PREVENTS THE RISE OF INTRACRANIAL PRESSURE DURING EXTRACORPOREAL RESUSCITATION IN SWINE.
Aim: Head and thorax elevation during cardiopulmonary resuscitation improves cerebral hemodynamics and ultimate neurological outcome after cardiac arrest. Its effect during extracorporeal cardiopulmonary resuscitation (E-CPR) is unknown. We tested whether this procedure could improve hemodynamics in swine treated by E-CPR. Methods and Results: Pigs were anesthetized and submitted to 15 minutes of untreated ventricular fibrillation followed by E-CPR. Animals randomly remained in flat position (flat group) or underwent head and thorax elevation since E-CPR institution (head-up group). Electric shocks were delivered after 30 minutes until the return of spontaneous circulation (ROSC). They were followed during 120 minutes after ROSC. After 30 minutes of E-CPR, ROSC was achieved in all animals, with no difference regarding blood pressure, heart rate, and extracorporeal membrane of oxygenation flow among groups. The head-up group had an attenuated increase in ICP as compared with the flat group after cardiac arrest (13 ± 1 vs. 26 ± 2 mm Hg at the end of the follow-up, respectively). Cerebral perfusion pressure tended to be higher in the head-up versus flat group despite not achieving statistical difference (66 ± 1 vs 46 ± 1 mm Hg at the end of the follow-up). Carotid blood flow and cerebral oxygen saturation were not significantly different among groups. Conclusion: During E-CPR, head and thorax elevation prevents ICP increase. Whether it could improve the ultimate neurological outcome in this situation deserves further investigation.


Selective methyl labeling of eukaryotic membrane proteins using cell-free expression.
Structural characterization of membrane proteins and other large proteins with NMR relies increasingly on perdeuteration combined with incorporation of specifically protonated amino acid moieties, such as methyl groups of isoleucines, valines, or leucines. The resulting proton dilution reduces dipolar broadening producing sharper resonance lines, ameliorates spectral crowding, and enables measuring of crucial distances between and to methyl groups. While incorporation of specific methyl labeling is now well established for bacterial expression using suitable precursors, corresponding methods are still lacking for cell-free expression, which is often the only choice for producing labeled eukaryotic membrane proteins in mg quantities. Here we show that we can express methyl-labeled human integral membrane proteins cost-effectively by cell-free expression based of crude hydrolyzed ILV-labeled OmpX inclusion bodies. These are obtained in Escherichia coli with very high quantity and represent an optimal intermediate to channel ILV precursors into the eukaryotic proteins.


Recent advances in the study of HPV-associated carcinogenesis.
Human papillomaviruses (HPVs) cause virtually all cervical cancers, the second leading cause of death by cancer among women, as well as other anogenital cancers and a subset of head and neck cancers. Approximately half of women, who develop cervical cancer die from it. Despite the optimism that has accompanied the introduction of prophylactic vaccines to prevent some HPV infections, the relatively modest uptake of the vaccine, especially in the developing world, and the very high fraction of men and women who are already infected, means that HPV-associated disease will remain as a significant public health problem for decades. In this review, we summarize some recent findings on HPV-associated carcinogenesis, such as miRNAs in HPV-associated cancers, implication of stem cells in the biology and therapy of HPV-positive cancers, HPV vaccines, targeted therapy of cervical cancer, and drug treatment for HPV-induced intraepithelial neoplasias.


Health effects associated with indoor swimming pools: a suspected toxic chloramine exposure.
A cohort and environmental study tested the hypothesis that suspected exposure to chloramines (by-products of chlorine for disinfection and ammonia from human sources) from a hotel's indoor swimming pool was associated with an outbreak among children who had attended a party at the pool. Retrospective cohort study. A case was defined as any hotel guest/visitor on 5-6 October 2002 who experienced three or more symptoms typical of chloramine exposure on either day after visiting the hotel. A cohort study and an environmental assessment were performed to determine the association between pool exposure and illness. Of 128 individuals interviewed, 32 met the case definition. Common symptoms among case patients were cough (84%), eye irritation (78%) and rash (34%). Illness was associated with entering the pool area [odds ratio (OR) 19.9; 95% confidence interval (CI) 2.3-172], but more strongly with swimming (OR 72.0; 95% CI 9.1-568). Pool chloramine levels on 6 October 2002 were > or = 0.7ppm (optimal level=0ppm; state maximum=0.5ppm). The pool operator lacked formal training in pool maintenance. High chloramine levels may have caused illness in individuals who were either in or near the pool. This outbreak underscores the need for regular pool maintenance, improved air quality, education and certification for all operators of public and semipublic pools, and education about healthy swimming practices.


Immediate cardiovascular effects of tension pneumothorax.
We studied the immediate cardiovascular effects of unilateral tension pneumothorax in sheep, a species with an intact mediastinal pleura similar to that in humans. Adult animals were restrained in the prone position and studied in the conscious state to permit spontaneous respiratory adjustments. We measured pressures in the ipsilateral pleural space (Ppli), contralateral pleural space (Pplc), esophagus (Pes), pulmonary artery (Ppa), superior vena cava (Pcv), carotid artery (Ps), as well as thermodilution cardiac output (CO) and heart rate (HR). Acute unilateral pneumothorax of 0, 10, and 20 cmH2O tension produced no significant changes in mean Ps, Pcv, and CO at any level of tension, but it did produce a marked increase in HR at 10 and 20 cmH2O tension (p less than 0.05). Similar observations were made with unilateral pneumothorax of 15 cmH2O for 30 min. These results suggested that tension was transmitted incompletely to the mediastinum and contralateral hemithorax. This was confirmed by an increase from baseline of mean (+/- SE) end-expiratory Pes by 6.6 +/- 4.9 cmH2O and Pplc by 6.7 +/- 6.0 cmH2O, for an increase in Ppli by 20.5 +/- 4.0 cmH2O. The respiratory intrathoracic pressure fluctuations were accentuated with mean increases of 211, 78, and 117% in the ipsilateral pleural space, esophagus, and contralateral pleural space, respectively (p less than 0.05). When the increase in respiratory intrathoracic pressure fluctuations was prevented by artificial hyperventilation, CO decreased. We conclude that unilateral tension pneumothorax does not directly lower CO and Ps because of (1) incomplete transmission of the ipsilateral pleural pressure to the mediastinum and contralateral hemithorax, (2) marked respiratory intrathoracic pressure swings, and (3) compensatory tachycardia.


Skin cancer incidence among atomic bomb survivors from 1958 to 1996.
The radiation risk of skin cancer by histological types has been evaluated in the atomic bomb survivors. We examined 80,158 of the 120,321 cohort members who had their radiation dose estimated by the latest dosimetry system (DS02). Potential skin tumors diagnosed from 1958 to 1996 were reviewed by a panel of pathologists, and radiation risk of the first primary skin cancer was analyzed by histological types using a Poisson regression model. A significant excess relative risk (ERR) of basal cell carcinoma (BCC) (n = 123) was estimated at 1 Gy (0.74, 95% confidence interval (CI): 0.26, 1.6) for those age 30 at exposure and age 70 at observation based on a linear-threshold model with a threshold dose of 0.63 Gy (95% CI: 0.32, 0.89) and a slope of 2.0 (95% CI: 0.69, 4.3). The estimated risks were 15, 5.7, 1.3 and 0.9 for age at exposure of 0-9, 10-19, 20-39, over 40 years, respectively, and the risk increased 11% with each one-year decrease in age at exposure. The ERR for squamous cell carcinoma (SCC) in situ (n = 64) using a linear model was estimated as 0.71 (95% CI: 0.063, 1.9). However, there were no significant dose responses for malignant melanoma (n = 10), SCC (n = 114), Paget disease (n = 10) or other skin cancers (n = 15). The significant linear radiation risk for BCC with a threshold at 0.63 Gy suggested that the basal cells of the epidermis had a threshold sensitivity to ionizing radiation, especially for young persons at the time of exposure.


The effects of interpersonal development programmes with sport coaches and parents on youth athlete outcomes: A systematic review and meta-analysis.
Interpersonal coach-and parent development programmes (CDP and PDP, respectively), have the goal to foster positive youth sport experiences through high-quality relations between coaches, parents, and youth athletes. In this paper we systematically reviewed the extant literature and estimate the overall magnitude of such programmes and how they can inform future interventions. Specifically, we aimed to: (a) conduct a systematic review on the literature of interpersonal CDPs and PDPs within the youth sport context; (b) examine the effects of such interventions on youth athlete outcomes via a meta-analysis. English written peer-reviewed publications and grey literature was identified through electronic search in databases and manual searches of reference lists. By utilising a priori criteria for inclusion and exclusion, 33 studies describing interpersonal CDPs, and PDPs were identified in the systematic review. Studies that presented required data for estimation of Hedge's g effect sizes were included in the meta-analysis (k = 27). By and large, the included studies used a quasi-experimental design (58%), sampled from team sports (79%), and reported several delivery methods (e.g., workshops, audio feedback, observations, peer group discussions) and outcome measures (e.g., anxiety, autonomous motivation, self-confidence). Some interventions were based on the same delivery protocols (e.g., Coach Effectiveness Training, Mastery Approach to Coaching) or theoretical frameworks (e.g., Achievement Goal Theory, Self-Determination Theory). The meta-analysis showed statistically significant small, and medium, effect sizes on a subsample of youth athlete outcomes (e.g., task-related climate, fun and enjoyment, anxiety), indicating that coach interpersonal skills can contribute to positive youth sport experiences. Theory-based interpersonal CDPs and PDPs are recommended to expand the knowledge in this field of research.


Investigations of fungal diversity in wooden structures and soils at historic sites on the Antarctic Peninsula.
Investigations of microbial diversity in Antarctic are important to begin to understand ecosystem functioning and decomposition processes. This study documents fungi at 9 historic sites on the Antarctic Peninsula collected from wooden structures, other organic materials, and soils during a joint National Science Foundation and British Antarctic Survey expedition in 2007. Many of these sites had wooden structures built by the British during the World War II Operation Tabarin, but others visited included the American "East Base" on Stonington Island and the Swedish hut on Snow Hill Island. Fungi were cultured on several different media and pure cultures were obtained and identified by DNA sequencing of the internal transcribed spacer region. Cadophora species previously found to attack historic wooden structures on Ross Island, Antarctica, were found at all but 1 location sampled in the Peninsula region. Fungi causing decay in the historic wooden structures and artifacts and those causing mold problems inside the structures are of great concern, and conservation efforts are urgently needed to help preserve these important polar heritage structures. The results presented also expand our knowledge on the identity of fungi present throughout the Antarctic Peninsula region and provide insights into the organisms responsible for decomposition and nutrient recycling.


Uncoupling interferons and the interferon signature explain clinical and transcriptional subsets in SLE.
Interferons (IFN) are thought to be key players in systemic lupus erythematosus (SLE). The unique and interactive roles of the different IFN families in SLE pathogenesis, however, remain poorly understood. Using reporter cells engineered to precisely quantify IFN-I, IFN-II and IFN-III activity levels in serum/plasma, we found that while IFNs play essential role in SLE pathogenesis and disease activity, they are only significant in specific subsets of patients. Interestingly, whereas IFN-I is the main IFN that governs disease activity in SLE, clinical subsets are defined by the co-elevation of IFN-II and IFN-III. Thus, increased IFN-I alone was only associated with cutaneous lupus. In contrast, systemic features, such as nephritis, were linked to co-elevation of IFN-I plus IFN-II and IFN-III, implying a synergistic effect of IFNs in severe SLE. Intriguingly, while increased IFN-I levels were strongly associated with IFN-induced gene expression (93.5%), in up to 64% of cases, the IFN signature was not associated with IFN-I. Importantly, neither IFN-II nor IFN-III explained IFN-induced gene expression in patients with normal IFN-I levels, and not every feature in SLE was associated with elevated IFNs, suggesting IFN-independent subsets in SLE. Together, the data suggest that, unlike the IFN signature, direct quantification of bioactive IFNs can identify pathogenic and clinically relevant SLE subsets amenable for precise anti-IFN therapies. Since IFN-I is only elevated in a subset of SLE patients expressing the IFN signature, this study explains the heterogeneous response in clinical trials targeting IFN-I, where patients were selected based on IFN-induced gene expression rather than IFN-I levels.


Determination of in vivo hepatic extraction ratio from in vitro metabolism by rat hepatocytes.
Using isolated rat hepatocytes, the in vitro intrinsic hepatic clearance of three drugs, namely, antipyrine (AP), propranolol (P), and CGS 13429 was determined. The hepatic extraction ratios of AP, P, and CGS 13429 were then calculated to be 0.10, 0.90, and 0.88, respectively. To determine the in vivo hepatic extraction ratio in the rat, AP, P, or CGS 13429 was infused separately at a rate of 40, 20, and 20 micrograms/min.kg, via jugular and hepatic portal veins. The steady state plasma levels of AP, P, and CGS 13429 were 5.30 +/- 0.33 micrograms/ml, 608 +/- 76 and 380 +/- 46 ng/ml after jugular vein (iv) infusions, and 4.96 +/- 0.46 micrograms/ml, 162 +/- 26, and 148 +/- 23 ng/ml after hepatic portal vein (hpv) infusions, respectively. The blood to plasma ratios of AP, P, and CGS 13429 were 1.0, 0.78, and 1.0, respectively. The in vivo hepatic extraction ratio, calculated as (Css,iv--Css,hpv)/Css,iv from steady state blood levels following iv and hpv infusions for AP, P, and CGS 13429, were 0.065, 0.73, and 0.61, respectively. These results suggest that in vitro metabolism studies by hepatocytes may have potential application, during drug discovery, to distinguish drugs of low and high metabolic hepatic extraction ratio in vivo.


Holistic bursting cells store long-term memory in auditory cortex.
The sensory neocortex has been suggested to be a substrate for long-term memory storage, yet which exact single cells could be specific candidates underlying such long-term memory storage remained neither known nor visible for over a century. Here, using a combination of day-by-day two-photon Ca2+ imaging and targeted single-cell loose-patch recording in an auditory associative learning paradigm with composite sounds in male mice, we reveal sparsely distributed neurons in layer 2/3 of auditory cortex emerged step-wise from quiescence into bursting mode, which then invariably expressed holistic information of the learned composite sounds, referred to as holistic bursting (HB) cells. Notably, it was not shuffled populations but the same sparse HB cells that embodied the behavioral relevance of the learned composite sounds, pinpointing HB cells as physiologically-defined single-cell candidates of an engram underlying long-term memory storage in auditory cortex.


Upper Airway Changes Following Different Orthognathic Surgeries, Evaluated by Cone Beam Computed Tomography: A Systematic Review and Meta-analysis.
This study sought to assess the upper airway changes following different orthognathic surgeries using cone-beam computed tomography. An electronic search of the literature was conducted in major electronic databases including Medline (PubMed), Web of Science, Scopus, and Open Grey for articles published up to January 20, 2018. Human studies that evaluated the changes in the volume and minimum cross-sectional area of the upper airway or its subdivisions in patients who had undergone orthognathic surgery by use of cone-beam computed tomography were included. Manual search of the bibliographies of the included articles was also conducted. The included studies underwent risk of bias assessment. A total of 1330 articles were retrieved. After excluding the duplicates and irrelevant articles, 41 studies fulfilled the eligibility criteria for this systematic review; out of which, 30 entered the meta-analysis. The majority of studies had a medium risk of bias. Mandibular setback, and maxillary advancement + mandibular setback decreased the volume of the upper airway (-6042.87 mm3 and -1498.78 mm3, respectively) and all its subdivisions in long-term (>3 months), except for the nasopharynx, the volume of which increased following maxillary advancement + mandibular setback. Mandibular advancement and maxillomandibular advancement both increased the upper airway volume in long-term (7559.38 mm3 and 7967.06 mm3, respectively); however, only the changes after the former procedure were significant. The minimum cross-sectional area increased after maxillomandibular advancement (161.43 mm2), and decreased following maxillary advancement + mandibular setback (-23.79 mm2) in long-term. There is moderate evidence to suggest that mandibular advancement is the only orthognathic movement that provides a statistically significant change in long-term upper airway volume.


MagFRET: the first genetically encoded fluorescent Mg2+ sensor.
Magnesium has important structural, catalytic and signaling roles in cells, yet few tools exist to image this metal ion in real time and at subcellular resolution. Here we report the first genetically encoded sensor for Mg(2+), MagFRET-1. This sensor is based on the high-affinity Mg(2+) binding domain of human centrin 3 (HsCen3), which undergoes a transition from a molten-globular apo form to a compactly-folded Mg(2+)-bound state. Fusion of Cerulean and Citrine fluorescent domains to the ends of HsCen3, yielded MagFRET-1, which combines a physiologically relevant Mg(2+) affinity (K d = 148 µM) with a 50% increase in emission ratio upon Mg(2+) binding due to a change in FRET efficiency between Cerulean and Citrine. Mutations in the metal binding sites yielded MagFRET variants whose Mg(2+) affinities were attenuated 2- to 100-fold relative to MagFRET-1, thus covering a broad range of Mg(2+) concentrations. In situ experiments in HEK293 cells showed that MagFRET-1 can be targeted to the cytosol and the nucleus. Clear responses to changes in extracellular Mg(2+) concentration were observed for MagFRET-1-expressing HEK293 cells when they were permeabilized with digitonin, whereas similar changes were not observed for intact cells. Although MagFRET-1 is also sensitive to Ca(2+), this affinity is sufficiently attenuated (K d of 10 µM) to make the sensor insensitive to known Ca(2+) stimuli in HEK293 cells. While the potential and limitations of the MagFRET sensors for intracellular Mg(2+) imaging need to be further established, we expect that these genetically encoded and ratiometric fluorescent Mg(2+) sensors could prove very useful in understanding intracellular Mg(2+) homeostasis and signaling.


Nerve Injury Increases the Expression of Alpha-2/Delta-1 Subunit of L-Type Calcium Channel in Sensory Neurons of Rat Spinal and Trigeminal Nerves.
By immunohistochemistry, an effect of nerve injury on distribution of alpha-2/delta-1 subunit of L-type calcium channel was investigated in rat's 4th and 5th lumbar dorsal root ganglia (DRGs), trigeminal ganglion (TG), and mesencephalic trigeminal nucleus (Mes5). The immunoreactivity was expressed by 52.2% of DRG neurons and 31.4% of TG neurons in intact animals. These neurons mostly had small-to-medium-sized cell bodies. In the DRG and TG, alpha-2/delta-1 subunit-positive neurons were lightly or moderately stained. However, the number of alpha-2/delta-1 subunit-immunoreactive (-IR) neurons dramatically increased in the ipsilateral DRG at 3-28 days after sciatic nerve transection (75.3-79.5%) and in the ipsilateral TG at 7 days after infraorbital nerve transection (66.3%). The IR density of alpha-2/delta-1 subunit in DRG and TG neurons was also elevated by the transection. In the injured DRG and TG, many sensory neurons with small-to-medium-sized cell bodies were strongly stained. Some large DRG and TG neurons showing strong staining intensity also appeared after the treatment. In the intact Mes5, sensory neurons were mostly devoid of alpha-2/delta-1 subunit-immunoreactivity (0.4%). However, alpha-2/delta-1-IR sensory neurons on the ipsilateral side of the Mes5 dramatically increased at 7 days after masseteric nerve transection (31.3%). A double immunofluorescence method also demonstrated that c-Jun activating transcription factor 3 (ATF3)-positive DRG (98.3-99.9%) and Mes5 (81.8%) neurons mostly co-expressed alpha-2/delta-1 subunit after the nerve injuries. However, alpha-2/delta-1 subunit immunoreactivity was relatively infrequent among ATF3-immunonegative DRG neurons (51.6-74.1%) and Mes5 neurons (<1%). The present study indicates that the nerve injury increases the protein level of alpha-2/delta-1 subunit among several types of axotomized sensory neurons in the spinal and trigeminal nervous systems.


Epidemiology of head lice infestation in primary school pupils, in khajeh city, East azerbaijan province, iran.
Pediculus capitis (Anoplura: Pediculidae) or head louse is an obligate ectoparasite transmitted mainly through physical contact. This study was conducted to survey the prevalence of head lice infestation rate and some risk factors in Primary School pupils, in Khajeh City East Azerbaijan Province, Iran We selected 20 primary schools of Khajeh City during 2008 and 2009. Totally 500 pupils including 200 boys and 300 girls from all grade 1-5 were selected by multistage, systematic random sampling in rural areas of Khajeh City and were examined for lice. In addition, a standard questionnaire recorded information about demographic features of each pupil. Results were analyzed by SPSS software. The total prevalence of head lice infestation in this study was 4.8%. and the prevalence rate was significantly higher in girls (6.66%) than in boys (2%). Epidemiological factors such as: sex, school grade, family size, parent's education, type of house, hair washing (per week), number of using comb per day, were evaluated and results showed significant difference in head lice infestation and sex, school grade, family size, father education, and type of house (P< 0.05). Pediculosis is a public health problem in many parts of the world, and due to the higher prevalence of pediculosis in crowded families, family by lower levels of father's education and socioeconomic status in our study and rural area, it is necessary to give health education for families to prevent of pediculosis in this area.


Frequency of Attainment of Low-Density Lipoprotein Cholesterol and Non-High-Density Lipoprotein Cholesterol Goals in Cardiovascular Clinical Practice (from the National Cardiovascular Data Registry PINNACLE Registry).
Studies have found that non-high-density lipoprotein cholesterol (non-HDL-C) is a superior marker for coronary heart disease compared to low-density lipoprotein cholesterol (LDL-C). Little is known about achievement of non-HDL-C goals outside clinical trials. Within a population of 146,064 patients with dyslipidemia in the PINNACLE Registry and a subgroup of 36,188 patients with diabetes mellitus (DM), we examined the proportion of patients and patient characteristics associated with having LDL-C, non-HDL-C, and both LDL-C and non-HDL-C levels at National Cholesterol Education Program goals. LDL-C, non-HDL-C, and both LDL-C and non-HDL-C goals in the overall cohort were achieved by 73%, 73.4%, and 68.9% patients, respectively. Significant predictors of meeting all 3 goals were age, male gender, statin, nonstatin, and combined statin plus nonstatin use. Patients with co-morbidities of hypertension, previous stroke or transient ischemic attack, peripheral arterial disease, myocardial infarction, and smoking were less likely to have LDL-C, non-HDL-C, and both LDL-C and non-HDL-C levels at National Cholesterol Education Program goal. In the overall cohort, patients with DM were less likely to meet non-HDL-C and both LDL-C and non-HDL-C goals. In the subgroup of patients with DM, predictors of meeting lipid goals were similar to the overall cohort. In conclusion, these data suggest contemporary treatment patterns by cardiologists successfully achieve lipid goals in most patients. Younger, female patients and those with atherosclerotic cardiovascular disease and risk factors, such as hypertension and DM, are less likely to achieve goals and may require more careful follow-up after statin initiation. Both LDL-C and non-HDL-C goals are achieved in <70% of patients, suggesting room for improvement if a goal-targeted individualized strategy is adopted.


Artificial Intelligence and Machine (Deep) Learning in Otorhinolaryngology: A Bibliometric Analysis Based on VOSviewer and CiteSpace.
Otorhinolaryngology diseases are well suited for artificial intelligence (AI)-based interpretation. The use of AI, particularly AI based on deep learning (DL), in the treatment of human diseases is becoming more and more popular. However, there are few bibliometric analyses that have systematically studied this field. The objective of this study was to visualize the research hot spots and trends of AI and DL in ENT diseases through bibliometric analysis to help researchers understand the future development of basic and clinical research. In all, 232 articles and reviews were retrieved from The Web of Science Core Collection. Using CiteSpace and VOSviewer software, countries, institutions, authors, references, and keywords in the field were visualized and examined. The majority of these papers came from 44 nations and 498 institutions, with China and the United States leading the way. Common diseases used by AI in ENT include otosclerosis, otitis media, nasal polyps, sinusitis, and so on. In the early years, research focused on the analysis of hearing and articulation disorders, and in recent years mainly on the diagnosis, localization, and grading of diseases. The analysis shows the periodical hot spots and development direction of AI and DL application in ENT diseases from the time dimension. The diagnosis and prognosis of otolaryngology diseases and the analysis of otolaryngology endoscopic images have been the focus of current research and the development trend of future.


Dual-Color Emissive Upconversion Nanocapsules for Differential Cancer Bioimaging In Vivo.
Early diagnosis of tumor malignancy is crucial for timely cancer treatment aimed at imparting desired clinical outcomes. The traditional fluorescence-based imaging is unfortunately faced with challenges such as low tissue penetration and background autofluorescence. Upconversion (UC)-based bioimaging can overcome these limitations as their excitation occurs at lower frequencies and the emission at higher frequencies. In this study, multifunctional silica-based nanocapsules were synthesized to encapsulate two distinct triplet-triplet annihilation UC chromophore pairs. Each nanocapsule emits different colors, blue or green, following a red light excitation. These nanocapsules were further conjugated with either antibodies or peptides to selectively target breast or colon cancer cells, respectively. Both in vitro and in vivo experimental results herein demonstrate cancer-specific and differential-color imaging from single wavelength excitation as well as far greater accumulation at targeted tumor sites than that due to the enhanced permeability and retention effect. This approach can be used to host a variety of chromophore pairs for various tumor-specific, color-coding scenarios and can be employed for diagnosis of a wide range of cancer types within the heterogeneous tumor microenvironment.


Quantitative analyses of the binding of soluble complement-fixing antibody/dsDNA immune complexes to CR1 on human red blood cells.
We have used direct binding isotherm analyses to measure the association constant (Ka) and number of binding sites for the binding of prepared complement-fixing antibody (Ab)/dsDNA immune complexes (IC) to human red blood cells (RBC). In order to generalize this study we have examined the binding reaction for a number of different anti-dsDNA Ab (from systemic lupus erythematosus plasmas), complement sources, RBC donors, and dsDNA sizes. The affinity of the IC for the RBC is quite high, and the Ka values fall within a narrow range (5 to 14 X 10(10) liter/mol). Similarly, the limiting stoichiometries for the number of IC bound per RBC were between 40 and 91. The very high affinity and limiting stoichiometries both suggest that the IC bind to the RBC via multiple contacts with clusters of complement receptor type 1 (CR1). Furthermore, we have used three specific monoclonal AB (mAb) to quantitate CR1 on human RBC in the presence and absence of bound IC. One of these Ab, mAb 1B4, is blocked from binding to the RBC if IC are previously bound, and we have used this observation to verify the multivalent nature of the interaction of complement-fixing IC with CR1 on human RBC.


Oldest horse brains: more advanced than previously realized.
Previous interpretations of early horse brains were based on an incorrectly identified fossil endocast, now believed to be from a condylarth. Newly prepared endocasts of Hyracotherium, the oldest horse and one of the earliest perissodactyls, reveal a relatively larger brain, with a more expanded neocortex, than existed in the condylarth ancestors of perissodactyls. Fifty million years ago, horse brains had suprasylvian, ectolateral, and lateral sulci, but the frontal lobe was undeveloped.


15N NMR relaxation studies of free and inhibitor-bound 4-oxalocrotonate tautomerase: backbone dynamics and entropy changes of an enzyme upon inhibitor binding.
The solution secondary structure of 4-oxalocrotonate tautomerase (4-OT), a 41 kDa homohexamer with 62 residues per subunit, consists of an alpha-helix, two beta-strands, a beta-hairpin, two loops, two turns, and a C-terminal coil [Stivers et al. (1996) Protein Sci. 5, 729-741]. The general base, proline-1, as well as the two loops and the beta-hairpin have been shown to comprise the active site [Stivers et al. (1996) Biochemistry 35, 814-823]. The backbone dynamics of both the free enzyme and its complex with a substrate analog have been studied by 1H-detected 15N relaxation rates and NOE determinations at 500 and 600 MHz. Analysis of the data using the model-free formalism showed that the nanosecond to picosecond motion of 53 of the 60 backbone 15N-H vectors was highly restricted with a mean order parameter mean value of S2 = 0.87 +/- 0.03. The lowest backbone mobility (S2 > 0.90) is found in the beta 1-strand, loop 2, and turn 2. Greater backbone mobility is found in the active site (0.5 < or = S2 < or = 0.83) and at C-terminal residues 58-62 (0.03 < or = S2 < or = 0.70). A tau m value for the free hexamer of 13.7 ns at 42 degrees C was determined, consistent with a compact globular molecule of 41 kDa. Saturation of 4-OT with the analog of the dienolic intermediate and linear competitive inhibitor cis, cis-muconate (4) (KD = 0.59 mM) increased the backbone S2 of seven residues and decreased the backbone S2 of another eight residues, both at the active site and at the antiparallel beta 1-beta 1 interface. The S2 values of the other 44 detectable NH vectors were not altered by the binding of 4. The increases in S2, resulting from the "freezing" of the backbone NH vectors of seven residues upon the binding of 4, correspond to an unfavorable entropic contribution to delta Gbinding of 3.2 +/- 1.1 kcal/mol. This freezing is partially compensated for by the mobilization of the other eight residues, since the decreases in S2 for these residues correspond to an entropic contribution to binding of -1.9 +/- 0.1 kcal/mol. These entropy changes, resulting solely from alterations in high-frequency motion, are significant compared to the overall delta Gbinding = -4.6 kcal/mol for 4. Other effects of the binding of 4 include (1) changes in 15N and NH chemical shifts localized to the active site and (2) increases in the exchange contributions (R(ex)) to 1/T2 of backbone 15N resonances at the active site and at the subunit interface, reflecting microsecond to millisecond motions which may play a role in substrate binding (k(on) > or = 4 x 10(6) M-1 s-1) and/or catalysis (kcat = 10(3) s-1).


Transcriptional up-regulation of the cyclin D2 gene and acquisition of new cyclin-dependent kinase partners in human T-cell leukemia virus type 1-infected cells.
Human T-cell leukemia virus type 1 (HTLV-1) is the etiologic agent for adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis. Tax(1) is a 40-kDa phosphoprotein, predominantly localized in the nucleus of the host cell, which functions to transactivate both viral and cellular promoters. It seems likely that HTLV-1, through expression of the viral regulatory protein Tax(1), provides some initial alteration in cell metabolism predisposing the development of ATL. Here, we demonstrate that HTLV-1 infection in T-cell lines and patient samples causes overexpression of an early G(1) cyclin, cyclin D2. The transcriptional up-regulation of the cyclin D2 gene is due to activation of Tax on the cyclin D2 gene. More important, we find that overexpression of cyclin D2 is accompanied by acquisition of new partners such as cyclin-dependent kinase 2 (cdk2), cdk4, and cdk6 in infected cells. This is in contrast to uninfected T cells, where cyclin D2 associates only with cdk6. Functional effects of these cyclin-cdk complexes in infected cells are shown by hyperphosphorylation of Rb and histone H1, indicators of active progression into S phase as well as changes in cellular chromatin and transcription machinery. These studies link HTLV-1 infection with changes of cellular cyclin gene expression, hence providing clues to development of T-cell leukemia.


Effects of a molasses-coated cottonseed product on diet digestibility, performance, and milk fatty acid profile of lactating dairy cattle.
An experiment was conducted to evaluate the effects of a molasses-coated cottonseed product on nutrient digestibility and milk fatty acid (FA) composition of lactating dairy cattle. The effect of a direct-fed microbial (DFM) product was also examined. Twelve Holstein cows (693+/-85kg of body weight, 127+/-39 d in milk, 2.08+/-0.29 lactations; mean +/- SD) were randomly assigned to sequence in a replicated 4 x 4 Latin square design balanced for carryover effects. Cows were fed 1 of 4 treatments during each of the four 14-d periods: a control diet including 11.4% (dry matter basis) reginned cottonseed (CON), a diet with 14.4% molasses-coated cottonseed to match the cottonseed inclusion rate of the control diet (TC), the control diet with the addition of a liquid form of the cotton coating used to produce molasses-coated cottonseed (LC), and the LC diet with the addition of a DFM (LC+DFM). Diets were formulated for equal concentrations of neutral detergent fiber, crude protein, ether extract, and macrominerals. Treatments had no effect on dry matter intake, apparent total-tract nutrient digestibility, or milk production. The molasses coat, in either form, tended to decrease concentrations of odd-chain FA (2.25 and 2.31 vs. 2.35 g/100g of FA for TC, LC, and CON, respectively) and unsaturated FA (31.4 and 31.1 vs. 32.1 g/100g of FA) in milk. This could be indicative of a mild shift in ruminal fermentation away from propionate-producing bacteria toward fiber-digesting bacteria responsible for biohydrogenation of FA. The form of the molasses coating had few effects, but LC significantly decreased concentrations of total trans-C18:1 (2.04 vs. 2.30+/-0.13 g/100g of FA) and polyunsaturated FA (4.81 vs. 5.01+/-0.17) compared with TC, implying that the liquid form slightly enhanced ruminal FA biohydrogenation. Furthermore, adding the DFM to the LC diet tended to increase the proportion of long-chain FA (FA >C16) and significantly increased the proportions of trans-C18:1 (2.22 vs. 2.04+/-0.13 g/100g of FA) and unsaturated FA (32.4 vs. 31.1+/-0.7 g/100g of FA), suggesting an inhibitory effect on ruminal biohydrogenation. Results suggest that coating cottonseed with a hardened molasses product does not significantly depress nutrient digestibility and may provide a convenient method of incorporating these ingredients into dairy rations.


Pelvic floor muscle training for bowel dysfunction following colorectal cancer surgery: A systematic review.
To identify, evaluate and synthesize the evidence examining the effectiveness of pelvic floor muscle training (PFMT) on bowel dysfunction in patients who have undergone colorectal cancer surgery. Eight electronic databases (MEDLINE 1950-2014; CINAHL 1982-2014; EMBASE 1980-2014; Scopus 1823-2014; PsycINFO 1806-2014; Web of Science 1970-2014; Cochrane Library 2014; PEDro 1999-2014) were systematically searched in March 2014. Reference lists of identified articles were cross referenced and hand searched. Randomized controlled trials, cohort studies and case series were included if they investigated the effects of conservative treatments, including PFMT on bowel function in patients with colorectal cancer following surgery. Two reviewers independently assessed the risk of bias of studies using the Newcastle-Ottawa Scale (NOS). Six prospective non-randomized studies and two retrospective studies were included. The mean (SD) NOS risk of bias score was 4.9 (1.2) out of 9; studies were limited by a lack of non-exposed cohort, lack of independent blinded assessment, heterogeneous treatment protocols, and lack of long-term follow-up. The majority of studies reported significant improvements in stool frequency, incontinence episodes, severity of fecal incontinence, and health-related quality of life (HRQoL) after PFMT. Meta-analysis was not possible due to lack of randomized controlled trials. Pelvic floor muscle training for patients following surgery for colorectal cancer appears to be associated with improvements in bowel function and HRQoL. Results from non-randomized studies are promising but randomized controlled trials with sufficient power are needed to confirm the effectiveness of PFMT in this population.


[Correlation between chronic idiopathic urticaria and infection due to H. pylori].
H. pylori is considered one of the major etiological agents for chronic idiopathic urticaria; it infects 15 percent of the world's population and induces an inflammatory process in the gastric mucosa. In some cases it is related to chronic urticaria, even in the absence of digestive symptoms. The purpose of this study is to find the correlation between chronic idiopathic urticaria and infection due to Helicobacter pylori. Twenty patients with chronic idiopathic urticaria and a positive urea breath test for H. pylori were included in the study. All other frequent pathologies were eliminated. Patients began treatment with amoxicillin, clarithromycin and omeprazole for 14 days. Thirty days after the start of treatment, the urea breath test was performed in all of the patients. Also cutaneous symptoms were evaluated. Urticaria disappeared in 11 patients (55%), in 9 (45%) the urea breath test became negative and in two remained positive. Urticaria persisted in 9 patients (45%), in 6 (30%) the urea breath test was negative and in 3 (15%) persisted as positive. Only two patients (10%) experienced gastric symptoms. We found a correlation between the disappearance of the symptoms and a negative urea breath test in 45 percent of the patients who were studied. We must consider the possibility of H. pylori infection, which can be present even without gastric symptoms.


Preparation of ferritin conjugates and antibodies for the localization and identification of antigens in tissues and cells by electron microscopy.
Ferritin, due to its high electron density, serves s a good marker in immunoelectron microscopic studies to localize antigens on and in cells and tissues. A step-by-step methodology for the conjugation of antibodies to ferritin is presented. Special attention has been placed on the techniques for the purification of ferritin and antibodies, conjugation procedures, and the treatment of specimens with conjugates. Methods for obtaining the separation of conjugated antibodies from unconjugated antibodies and for testing the purity and specificity of the labeling reagents are discussed.


The relationships between marijuana use and exercise among young and middle-aged adults.
As decriminalization, increased access, and decreased perception of risk spurs the popularity of medical and recreational marijuana, more information is needed on possible links between marijuana use and social determinants of health. The goal of this study was to assess the relationships between marijuana use and exercise. Data sources included the two most recent waves of the National Longitudinal Study of Adolescent to Adult Health (Wave IV, 2008-2009; N = 14,784 and Wave V, 2016-2018; N = 12,043). The exercise variables included any form of exercise/sport during the past 7 days and the number of days participating in each of 7 types of exercise/sport. Marijuana-use variables comprised any current use and frequency of use during the past 30 days. Both fixed-effects and random-effects models were estimated with numerous control variables, along with binary and count measures of exercise. Results show that, particularly for fixed-effects models, marijuana use is not significantly related to exercise, counter to conventional wisdom that marijuana users are less likely to be active. Indeed, the only significant estimates suggest a positive relationship, even among heavier users during the past 30 days. These findings are at odds with much of the existing literature, which generally shows a negative relationship between marijuana use and exercise. As additional states legalize the medicinal and recreational use of marijuana, perhaps its impact on exercise, one of the leading social determinants of health, is not necessarily a primary concern.