miR-669a-5p promotes adipogenic differentiation and induces browning in preadipocytes.
Obesity is a major global health issue that contributes to the occurrence of metabolic disorders. Based on this fact, understanding the underlying mechanisms and to uncover promising therapeutic approaches for obesity have attracted intense investigation. Brown adipose tissue (BAT) can help burns excess calories. Therefore, promoting White adipose tissue (WAT) browning and BAT activation is an attractive strategy for obesity treatment. MicroRNAs (miRNAs) are small, non-coding RNAs, which are involved in regulation of adipogenic processes and metabolic functions. Evidence is accumulating that miRNAs are important regulators for both brown adipocyte differentiation and white adipocyte browning. Here we report that the expression of miR-669a-5p increases during the adipogenic differentiation of 3T3-L1 and C3H10T1/2 adipocytes. miR-669a-5p supplementation promotes adipogenic differentiation and causes browning of 3T3-L1 and C3H10T1/2 cells. Moreover, the expression of miR-669a-5p is upregulated in iWAT of mice exposed to cold. These data demonstrate that miR-669a-5p plays a role in regulating adipocyte differentiation and fat browning.Abbreviations: Acadl: long-chain acyl-Coenzyme A dehydrogenase; Acadm: medium-chain acyl-Coenzyme A dehydrogenase; Acadvl: very long-chain acyl-Coenzyme A dehydrogenase, very long chain; Aco2: mitochondrial aconitase 2; BAT: brown adipose tissue; Bmper: BMP-binding endothelial regulator; Cpt1-b:carnitine palmitoyltransferase 1b; Cpt2: carnitine palmitoyltransferase 2; Crat: carnitine acetyltransferase; Cs: citrate synthase; C2MC: Chromosome 2 miRNA cluster; DMEM: Dulbecco's modified Eagle medium; eWAT: epididymal white adipose tissue; ETC: electron transport chain; FAO: fatty acid oxidation; Fabp4:fatty acid binding protein 4; FBS: fetal bovine serum; Hadha: hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha; Hadhb: hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit beta; HFD: high fat diet; Idh3a: isocitrate dehydrogenase 3 alpha; iWAT: inguinal subcutaneous white adipose tissue; Lpl: lipoprotein lipase; Mdh2: malate dehydrogenase 2; NBCS: NewBorn Calf Serum; mt-Nd1: mitochondrial NADH dehydrogenase 1; Ndufb8:ubiquinone oxidoreductase subunit B8; Nrf1: nuclear respiratory factor 1; Pgc1α: peroxisome proliferative activated receptor gamma coactivator 1 alpha; Pgc1b: peroxisome proliferative activated receptor, gamma, coactivator 1 beta; Pparγ: peroxisome proliferator activated receptor gamma; Prdm16: PR domain containing 16; Rgs4: regulator of G-protein signaling 4; Sdhb: succinate dehydrogenase complex, subunit B; Sdhc: succinate dehydrogenase complex, subunit C; Sdhd: succinate dehydrogenase complex, subunit D; Sh3d21: SH3 domain containing 21; Sfmbt2: Scm-like with four mbt domains 2; TG: triglyceride; TCA: tricarboxylic acid cycle; Tfam: transcription factor A, mitochondrial; TMRE: tetramethylrhodamine, methyl ester; Ucp1: uncoupling protein 1; Uqcrc2: ubiquinol cytochrome c reductase core protein 2; WAT: White adipose tissue.


[Loss of total 5-methylcytosine from the genome during cell culture aging coincides with the Hayflick limit].
Analyzing the data about the age-related 5-methylcytosine (5mC) loss from DNA of cell cultures, the following conclusions have been made: 1. The rate of 5mC loss from DNA does not depend on the cell donor age; it remains constant during the logarithmic phase of cell growth, and may vary significantly in different cell lines. 2. The rate is inversely proportional to their Hayflick limit and to the species lifespan of cell donors. 3. In immortal cell lines the 5mC content in DNA is stable or increases with aging. 4. Hayflick limit estimations coincide with or are lower than the number of cell population doublings that corresponds to all 5mC loss from cell genome. A simple and fast method has been proposed for Hayflick limit prognostication by analysis of the rate of DNA hypomethylation. It may be used for early diagnosis of precrisis and immortal cell lines. Evidence has been obtained that age-dependent 5mC loss from DNA is the result of accumulating 5mC-->T+C substitutions that occur during DNA methylation in every cell division. The loss of all genomic 5mC residues during the lifespan may correspond to accumulation of about 3 x 10(6) 5mC-->T transitions or, on average, one mutation per gene. This may be one of the main reasons of the "catastrophe of errors" and cessation of cell proliferation. It is calculated that the rate of 5mC-->T transitions in normal cells may be 2.3 x 10(-5) per site in each cell doubling in human, 6 x 10(-5) in hamster, and 4.6 x 10(-4) in mouse. DNA methylation as a generator of mutations may be a "counter" of cell divisions and thus be one of the molecular mechanisms of the Hayflick phenomenon. The conclusion is made that the DNA methylation system may be considered as a genetically programmed mechanism for accumulating mutations during cell aging.


Why Do People Think Individuals in Poverty Are Less Vulnerable to Harm?: Testing the Role of Intuitions About Adaptation.
People often falsely believe that individuals from low socioeconomic status (SES) backgrounds are less harmed than those from higher SES backgrounds by a wide range of negative events. We report three studies (total N = 1,625) that provide evidence that this "thick skin bias" emerges at least in part because people overgeneralize otherwise accurate intuitions about adaptation. Across studies, participants accurately intuited that people adapt to psychophysical experiences (e.g., brightness, weight, and volume) but also inaccurately intuited that people similarly adapt to life hardships that actually tend to exacerbate the harm of future negative events. Experimentally decreasing the salience of psychophysical adaptation intuitions reduced the thick skin bias, suggesting a causal link between these adaptation intuitions and the belief that people in poverty are less vulnerable to harm and underlining the importance of studying how biased beliefs about the effects of poverty may perpetuate inequality.


Online versus offline processing of visual feedback in the control of movement amplitude.
Researchers have suggested that visual feedback not only plays a role in the correction of errors during movement execution but that visual feedback from a completed movement is processed offline to improve programming on upcoming trials. In the present study, we examined the potential contribution of online and offline processing of visual feedback by analysing spatial variability at various kinematic landmarks in the limb trajectory (peak acceleration, peak velocity, peak negative acceleration and movement end). Participants performed a single degree of freedom video aiming task with and without vision of the cursor under four criterion movement times (225, 300, 375 and 450 ms). For movement times of 225 and 300 ms, the full vision condition was less variable than the no vision condition. However, the form of the variability profiles did not differ between visual conditions suggesting that the contribution of visual feedback was due to offline processes. In the 375 and 450 ms conditions, there was evidence for both online and offline control as the form of the variability profiles differed significantly between visual conditions.


Enhancement of neurogenesis and memory by a neurotrophic peptide in mild to moderate traumatic brain injury.
Traumatic brain injury (TBI) is a risk factor for Alzheimer disease (AD), a neurocognitive disorder with similar cellular abnormalities. We recently discovered a small molecule (Peptide 6) corresponding to an active region of human ciliary neurotrophic factor, with neurogenic and neurotrophic properties in mouse models of AD and Down syndrome. To describe hippocampal abnormalities in a mouse model of mild to moderate TBI and their reversal by Peptide 6. TBI was induced in adult C57Bl6 mice using controlled cortical impact with 1.5 mm of cortical penetration. The animals were treated with 50 nmol/d of Peptide 6 or saline solution for 30 days. Dentate gyrus neurogenesis, dendritic and synaptic density, and AD biomarkers were quantitatively analyzed, and behavioral tests were performed. Ipsilateral neuronal loss in CA1 and the parietal cortex and increase in Alzheimer-type hyperphosphorylated tau and A-β were seen in TBI mice. Compared with saline solution, Peptide 6 treatment increased the number of newborn neurons, but not uncommitted progenitor cells, in dentate gyrus by 80%. Peptide 6 treatment also reversed TBI-induced dendritic and synaptic density loss while increasing activity in tri-synaptic hippocampal circuitry, ultimately leading to improvement in memory recall on behavioral testing. Long-term treatment with Peptide 6 enhances the pool of newborn neurons in the dentate gyrus, prevents neuronal loss in CA1 and parietal cortex, preserves the dendritic and synaptic architecture in the hippocampus, and improves performance on a hippocampus-dependent memory task in TBI mice. These findings necessitate further inquiry into the therapeutic potential of small molecules based on neurotrophic factors.


Evolution of visual acuity, flap thickness, and optical density after laser in situ keratomileusis performed with a femtosecond laser.
To describe the postoperative evolution of visual acuity, flap morphology, and stromal optical density femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK). Clínica Rementería, Madrid, Spain. Prospective case series. The study comprised 44 consecutive eyes that had FS-LASIK performed with the WaveLight FS200 and the Allegretto excimer laser to correct myopia. Visual outcomes, flap thickness, and stromal optical density were assessed 1 day, 1 week, 1 month, and 3 months postoperatively. A statistically significant improvement in the mean uncorrected distance visual acuity at 1 day (0.94 ± 0.2) and 1 week (0.93 ± 0.2) to 1 month (1.04 ± 0.2) and 3 months (1.11 ± 0.1) postoperatively (P < .05). At 3 months postoperatively, the femtosecond laser had good outcomes in efficacy (0.98 ± 0.1), safety (0.98 ± 0.1), and predictability (100% of eyes were within ± 0.5 D of emmetropia). The femtosecond-created flaps were slightly thicker than intended, the mean SD intraflap was 7.1 μm, the range between the thickest and thinnest points in each flap was 25.4 μm, and the mean flap thickness homogeneity was 7.6 μm at 3 months postoperatively. A progressive decrease in the optical density of the flap stroma and the residual stromal bed was detected during follow-up. The femtosecond laser study appears to be a safe, effective, and predictable platform to obtain LASIK flaps. The flaps were planar and homogeneous but slightly thicker than intended. The optical density of the flap stroma was slightly higher at early follow-up and decreased over time.


Burnout among Danish prison personnel: A question of quantitative and emotional demands.
Objectives The objective of this follow-up study was to investigate associations between individual, occupational and work environment factors and burnout among both uniformed and non-uniformed personnel working in the Danish Prison and Probation Service. Methods The participants (N = 4808) with client contact received a questionnaire in 2010 and again in 2011. In 2010, 2843 participants responded to the questionnaire (59.1%), and in 2011, 1741 responded to the questionnaire, yielding a response rate of 61.2% of the baseline population, and 36.2% of the invited population. Burnout and work characteristics were measured with validated scales from the Copenhagen Psychosocial Questionnaire, and data was analysed by logistic regression. Results Risk factors with the highest impact on burnout were work environmental factors: quantitative demands, emotional demands, involvement in and meaning of work. Role conflict, role clarity, social support and demands for hiding emotions had borderline significance. Besides cohabitation, there was no association between individual factors and burnout or between occupational characteristics and burnout. Moreover, there was no association between exposure to threats and violence and burnout. Efforts for preventing burnout ought to be concentrated on reducing the quantitative job-demands, on easing and improving staff-inmate relationships, but also on involvement in and meaning of work. Most work in prison is invisible and the overall goals are in conflict with each other. Management must provide solutions to problems of role conflict and support groups for social support. There is a risk of burnout among both uniformed personnel and non-uniformed personnel working in both open and closed facilities.


[Anti-C. trachomatis IgG antibodies levels in serum to identify infertility caused by tubal damage].
To identify the serologic titers of anti-Chlamydia trachomatis IgG (Ab) antibodies that could be used to differentiate tubal damage infertility from other causes of subfertility in a group of Mexican women. This was a prospective, longitudinal and analytical study of 147 women selected in a non-random way. The women were classified into three sub-groups: 1) infertile women with tubal occlusion detected by laparoscopy (n = 58); 2) infertile women with alternative causes of subfertility (n = 50), and 3) fertile women for the control group (n = 39). An assay of indirect immunofluorescence was performed on all infertile women (n = 108). The results obtained were compared with the laparoscopic and hormonal analyses carried out on the 108 infertile women. The statistical analysis included a model in ROC Curve and Logistical Regression. The results showed that the titer 1:256 is able to differentiate fertile women from infertile women. Moreover, in the adjusted analysis, the titer 1:512 was able to identify infertile women with tubal occlusion (OR 2.6, CI 95% 1.24, 5.4), with a sensibility of 40% and a specificity of 90%. Positive and negative predictive values were 85% and 50%, respectively and the positive and negative likelihood ratios were 3.85 and 0.67, respectively. The pattern of the ROC curve confirmed a court value of 1:512, with an area under the curve of 62.2% (CI 95%: 53.4-72%). A titer greater or equal to 1:512 of anti-C. trachomatis IgG antibodies is useful in the identification of tubal factor infertility.


Phenotypes, genotypes, and prevalence of congenital myopathies older than 5 years in Denmark.
Congenital myopathy as a nosologic entity has long been recognized, but knowledge of overall and subtype prevalence and phenotype-genotype relationship is scarce, especially in the adult population. A national cohort of 107 patients ≥5 years diagnosed with congenital myopathy were prospectively assessed clinically, histologically, and genetically. Twenty-five patients were excluded because of atypical features or alternative etiologies. The remaining 82 were on average 28 years old. Histologic examination revealed 14 (17%) with core disease, 15 (18%) centronuclear myopathy, 12 (15%) nemaline rods, 27 (33%) congenital fiber-type disproportion or type I predominance, and 14 (17%) nonspecific myopathic changes. Genetic etiology was identified in 46 patients (56.1%); 22.0% were heterozygous or compound heterozygous for mutations in RYR1, 7.3% had DNM2 mutations, and 7.3% NEB mutations. Less than 5% had mutations in ACTA1, TPM2/3, MTM1, TTN, SEPN1, or SC4NA. A genetic cause was established in 83% with specific histology (cores/rods/centronuclear myopathy) vs 29% with unspecific histology. The detailed clinical examination found gene-dependent discrepancies in the pattern of muscle affection and walking ability. Although walking ability was delayed in patients with ACTA1, TPM2/3, and RYR1 mutations, it was within normal limits in patients with NEB and DNM2 mutations. We found that overall, genetic and histologic prevalence of congenital myopathy in Denmark differs from previous retrospective reports. Less RYR1 and more DNM2 and NEB mutations and less core histology were present in our cohort. These differences may be explained by our prospective design, the older cohort of patients, and by differences in genetic background.


Readmission and processes of care across weekend and weekday hospitalisation for acute myocardial infarction, heart failure or stroke: an observational study of the National Readmission Database.
Variation in hospital resource allocations across weekdays and weekends have led to studies of the 'weekend effect' for ST elevation myocardial infarction (STEMI), non-ST elevation myocardial infarction (NSTEMI), heart failure (HF) and stroke. However, few studies have explored the 'weekend effect' on unplanned readmission. We aimed to investigate 30-day unplanned readmissions and processes of care across weekend and weekday hospitalisations for STEMI, NSTEMI, HF and stroke. We grouped hospitalisations for STEMI, NSTEMI, HF or stroke into weekday or weekend admissions. Multivariable adjusted ORs for binary outcomes across weekend versus weekday (reference) groups were estimated using logistic regression. We included all non-elective hospitalisations for STEMI, NSTEMI, HF or stroke, which were recorded in the US Nationwide Readmissions Database between 2010 and 2014. The analysis sample included 659 906 hospitalisations for STEMI, 1 420 600 hospitalisations for NSTEMI, 3 027 699 hospitalisations for HF, and 2 574 168 hospitalisations for stroke. The primary outcome was unplanned 30-day readmission. As secondary outcomes, we considered length of stay and the following processes of care: coronary angiography, primary percutaneous coronary intervention, coronary artery bypass graft, thrombolysis, brain scan/imaging, thrombectomy, echocardiography and cardiac resynchronisation therapy/implantable cardioverter-defibrillator. Unplanned 30-day readmission rates were 11.0%, 15.1%, 23.0% and 10.9% for STEMI, NSTEMI, HF and stroke, respectively. Weekend hospitalisations for HF were associated with a statistically significant but modest increase in 30-day readmissions (OR of 1.045, 95% CI 1.033 to 1.058). Weekend hospitalisation for STEMI, NSTEMI or stroke was not associated with increased risk of 30-day readmission. There was no clinically meaningful evidence against the supposition that weekend and weekday hospitalisations have the same 30-day unplanned readmissions. Thirty-day readmission rates were high, especially for HF, which has implications for service provision. Strategies to reduce readmission rates should be explored, regardless of day of hospitalisation.


Identification of fetal gender in maternal blood is a helpful tool in the prenatal diagnosis of haemophilia.
Fetal DNA identification in maternal circulation has provided a new approach for non-invasive prenatal diagnosis. However, fetal DNA can persist in maternal blood long after the delivery, severely hampering this possibility. We addressed the issue of fetal DNA persistence in maternal blood. Thus, we investigated cell-free fetal DNA as a reliable approach in prenatal diagnosis of haemophilia. Forty non-pregnant women, who had had at least a male fetus, 29 control pregnant women, and 14 pregnant women, carriers of hemophilia A or B. The assessment of Y-chromosomal sequences was performed by analysing SRY and amelogenin genes using PCR-based techniques. A protocol consisting of double centrifugation at full speed followed by plasma filtration hampered the detection of Y chromosome-specific sequence in non-pregnant women. In 29 control pregnant women, blinded determination of fetal sex confirmed the specificity and sensitivity of the method applied. In 14 pregnant carriers of hemophilia, the investigation revealed a male fetus in nine pregnancies. Excluding the three cases in which a spontaneous miscarriage occurred, the sensitivity and specificity of fetal sex prediction by SRY and amelogenin gene analyses were both 100% as compared with the invasive approach and the fetal sex outcome at birth (six males and five females). Because of its high accuracy in prediction, fetal gender determination with cell-free fetal DNA in maternal plasma may be a useful tool in prenatal diagnosis of haemophilia allowing for the avoidance of invasive procedures for female fetuses.


Comparison of high-dose therapy and autologous stem-cell transplantation with conventional therapy for Hodgkin's disease induction failure: a case-control study. Société Francaise de Greffe de Moelle.
To determine the prognostic factors and outcome of first-line induction failure Hodgkin's disease patients who were treated with a salvage regimen of high-dose chemotherapy and autologous stem-cell transplantation, and to compare them with matched, conventionally treated patients. We retrospectively analyzed data relating to 86 Hodgkin's disease patients who underwent autologous stem-cell transplantation after failure of the first chemotherapy regimen, either because they did not enter a complete remission and experienced progression of disease less than 3 months after the end of their first-line treatment or because they showed evidence of disease progression during first-line therapy. Graft patients were matched with 258 conventionally treated patients (three controls per case) for age, sex, clinical stage, B symptoms, and time at risk; patient data were obtained from international databases. Among the 86 graft patients, the median age at diagnosis was 29 years (range, 14 to 57 years). Thirty-nine percent of patients had stage II disease, 23% had stage III disease, and 38% had stage IV disease. Seventy percent of the patients received chemotherapy and 30% received combined modality therapy; 60% of the patients received a seven- or eight-drug regimen. After this first-line treatment, 91% had disease progression and 9% had a brief partial response. Eighty patients received a second-line treatment; pretransplantation status was as follows: 24% of patients had a complete remission, 38% had a partial remission (PR), 14% had stable disease, and disease progression occurred in 24%. With a median follow-up of 22 months (range, 4 to 105 months) from diagnosis, the 5-year event-free survival and overall survival rates from transplantation were 25% and 35% (95% confidence intervals, 15 to 36 and 23 to 49), respectively. In multivariate analysis, the pretransplantation disease status after salvage therapy was the only significant prognostic factor for survival (PR: relative risk = 2.8, P = .017; progressive disease: relative risk (RR) = 5.26, P < .001). From diagnosis, the 6-year overall survival rates of the graft patients and 258 matched conventionally treated patients were 38% and 29%, respectively (P = .058). Autologous stem-cell transplantation represents the best therapeutic option currently available for patients with primary induction failure and is associated with acceptable toxicity. Response to second-line treatment before high-dose chemotherapy is the only prognostic factor that can be correlated with survival.


Improvement in relaxation by nifedipine in hypoxic isometric cat papillary muscle.
Hypoxia has been demonstrated to cause impairment of myocardial relaxation. This impairment of relaxation is particularly pronounced during early reoxygenation. This study was undertaken in 24 isometric cat papillary muscles at 38 degrees C to determine if nifedipine can influence the impairment of relaxation produced during reoxygenation following hypoxia. A dose of nifedipine was chosen that produced only a minimal depression of peak systolic tension and no change in the half time to relaxation (RT 1/2) under well-oxygenated conditions. Thirty minutes of hypoxia were produced in 12 muscles, and systolic tension decreased by the same amount in muscles treated or not treated with nifedipine. During early hypoxia in the absence of nifedipine, RT1/2 was significantly prolonged (P less than 0.01) from 104 +/- 7 to 126 +/- 9 ms. After pretreatment with nifedipine, the change in RT1/2 with hypoxia was not significant. More striking was the near abolition of the marked impairment of relaxation seen during early reoxygenation (238 +/- 33 ms without nifedipine and 128 +/- 8 ms with nifedipine, P less than 0.01). These data establish that, although nifedipine only minimally attenuates the relaxation impairment early during hypoxia, this agent can substantially reduce the impairment of relaxation produced by early reoxygenation.


Targeting HIF-1α alleviates the inflammatory responses and rebuilds the CD4+ T cell subsets balance in the experimental autoimmune myasthenia gravis inflammation model via regulating cellular and humoral immunity.
Cells and tissues in an inflammatory state are usually hypoxic. The hypoxic environment can affect the differentiation of immune cells and produce Hypoxia-inducible Factor-1α (HIF-1α). Inflammation is also a major contributor to the development and deterioration of Myasthenia Gravis (MG). There are limited studies on the immunopathological mechanism and targeted therapy associated with MG exacerbated with inflammation. This research aimed to explore whether BAY 87-2243 (HIF-1α inhibitor) ameliorates the symptoms of the Experimental Autoimmune Myasthenia Gravis (EAMG) inflammation model and study its regulatory mechanism on cellular immunity and humoral immunity. We first establish the EAMG inflammation model using Lipopolysaccharide (LPS), BAY 87-2243 was applied to the EAMG inflammation model and its therapeutic effects were evaluated in vivo and in vitro experiments. The proportion of Treg cells was increased whereas Th1, Th17, and Th1/17 cells were decreased in BAY 87-2243-treated EAMG inflammation model. BAY 87-2243 ameliorated the acetylcholine receptors (AChRs) loss and the complement deposited at the neuromuscular junction of the EAMG inflammation model, declined the levels of IFN-γ, IL-17, and IL-6 in serum, and further attenuated responses in the germinal center and reduced the antibody levels by inhibiting the IL-6-dependent STAT3 axis. BAY 87-2243 restored the balance of CD4+T cell subsets and reduced the production of the pro-inflammatory cytokines, thus acting as both an immune imbalance regulator and anti-inflammatory. The current study suggests that HIF-1α might be a potential target for the treatment of MG exacerbated with inflammation.


The role of interferon-gamma and interferon-gamma receptor in tuberculosis and nontuberculous mycobacterial infections.
Mycobacterium tuberculosis (Mtb) and nontuberculous mycobacteria (NTM) remain the leading causes of lung disease and mortality worldwide. Interferon-gamma (IFN-γ) and its receptor (IFN-γR) play a key role in mediating immunity against Mtb and NTM. This study was conducted as a systematic review; all information was collected from databases such as: PubMed, Scopus, Medline, SID, and medical databases. Finally, all the collected data were reviewed, and all content was categorized briefly. There is growing evidence that IFN-γ plays an important role in host defense against these two intracellular pathogens by activating macrophages. In addition, IFN-γ has been shown to be an integral part of various antibacterial methods such as granuloma formation and phagosome-lysosome fusion, both of which lead to the death of intracellular Mycobacterium. As a result, its absence is associated with overgrowth of intracellular pathogens and disease caused by Mtb or Mycobacterium nontuberculosis. We also look at the role of IFN-γR in Mtb or NTM because IFN-γ acts through IFN-γR. Finally, we introduce new approaches to the treatment of M. tuberculosis complex (MTC) and NTM disease, such as cell and gene-based therapies that work by modulating IFN-γ and IFN-γR.


The use of the synthetic peptide, Gly-Pro-Arg-Pro, in the preparation of thrombin-degranulated rabbit platelets.
The method for preparing thrombin-degranulated platelets has been modified to avoid the use of plasmin or successive treatments with small amounts of thrombin, while still achieving more than 90% release of platelet amine storage granule contents. It was necessary to prevent the fibrinogen released from the platelets during thrombin treatment from forming an insoluble fibrin mesh that could trap the platelets and hinder their deaggregation. To accomplish this we have treated rabbit platelets with 0.73 U/ml of thrombin for 1 min in the presence of the synthetic peptide, Gly-Pro-Arg-Pro, which prevents the polymerization of fibrin molecules. We have demonstrated that it also prevents 125I, initially added as 125I-fibrinogen, from associating with the platelets in a form that was not removed by centrifuging and washing during the preparation of thrombin-degranulated platelets, and we infer that products formed from the fibrinogen released from the platelets would also be prevented from associating with them. Thrombin-degranulated platelets prepared by this method have lost 92% of their granule contents and they can be washed and resuspended. These platelets aggregate normally upon stimulation with thrombin, adenosine diphosphate (ADP), or arachidonate. Thus, Gly-Pro-Arg-Pro is useful in preparing thrombin-degranulated platelets for studying platelet reactions without the complicating effects of released materials such a ADP and fibrinogen.


Prevention of mother to child transmission of HIV.
Perinatal transmission is the most common cause of HIV infection in pediatric population below the age of 15 years. Much progress has been made to decrease the risk of transmission than just offering the option of medical termination of pregnancy to the mother. Future research is needed not only to develop the simple preventive program, but also to make it more cost effective, acceptable and accessible to the general population. Following review article focuses on the factors affecting the transmission of HIV, strategies for prevention of MTCT with special reference to use of nevirapine and breast-feeding practices in HIV positive mother. The role of counseling and voluntary testing is also stressed upon.


Intersection of small RNA pathways in Arabidopsis thaliana sub-nuclear domains.
In Arabidopsis thaliana, functionally diverse small RNA (smRNA) pathways bring about decreased RNA accumulation of target genes via several different mechanisms. Cytological experiments have suggested that the processing of microRNAs (miRNAs) and heterochromatic small interfering RNAs (hc-siRNAs) occurs within a specific nuclear domain that can present Cajal Body (CB) characteristics. It is unclear whether single or multiple smRNA-related domains are found within the same CB and how specialization of the smRNA pathways is determined within this specific sub-compartment. To ascertain whether nuclear smRNA centers are spatially related, we localized key proteins required for siRNA or miRNA biogenesis by immunofluorescence analysis. The intranuclear distribution of the proteins revealed that hc-siRNA, miRNA and trans-acting siRNA (ta-siRNA) pathway proteins accumulate and colocalize within a sub-nuclear structure in the nucleolar periphery. Furthermore, colocalization of miRNA- and siRNA-pathway members with CB markers, and reduced wild-type localization patterns in CB mutants indicates that proper nuclear localization of these proteins requires CB integrity. We hypothesize that these nuclear domains could be important for RNA silencing and may partially explain the functional redundancies and interactions among components of the same protein family. The CB may be the place in the nucleus where Dicer-generated smRNA precursors are processed and assigned to a specific pathway, and where storage, recycling or assembly of RNA interference components takes place.


Thrombosis in systemic lupus erythematosus: striking association with the presence of circulating lupus anticoagulant.
The lupus anticoagulant was found in the plasma of 31 of 60 patients with systemic lupus erythematosus and other connective tissue disorders (mixed connective tissue disease, systemic vasculitis, polyarteritis nodosa, primary sicca syndrome, discoid lupus, Behcet's syndrome, and systemic sclerosis). Strong associations were found with biological false positive seroreaction for syphilis and thrombocytopenia. The most striking association, however, was with the high prevalence of thrombosis. This tendency to thrombosis was independent of disease activity of systemic lupus erythematosus. The lupus anticoagulant appears to be a useful marker for a subset of patients with systemic lupus erythematosus at risk for the development of thromboembolic complications.


MAP2 is a component of crossbridges between microtubules and neurofilaments in the neuronal cytoskeleton: quick-freeze, deep-etch immunoelectron microscopy and reconstitution studies.
Microtubules (MT) and neurofilaments (NF) are linked by frequent crossbridges in situ. In order to answer the question of what makes these crossbridges, we performed the immunogold procedure on rat spinal cord motor neurons using an affinity-purified polyclonal antibody against rat brain MAP2 and gold-labeled anti-rabbit IgG goat IgG. A quick-freeze, deep-etch technique (QF-DE) in conjunction with decoration with anti-MAP2 antibody and ferritin-labeled second antibody was also used. In motor neuron dendrites crossbridges were clearly displayed between MTs and NFs by QF-DE. These crossbridges were revealed in thin sections as fuzzy filamentous structures between MT and NF. Gold particles studded the fuzzy structures associated with MT. Many such structures connected MTs to NFs. Furthermore, antibody complexes containing ferritin were localized on the crossbridges between MTs and NFs by the QF-DE study. In addition, we performed reconstitution experiments. We isolated 70 kDa (L) protein of neurofilaments from calf spinal cords and assembled L to form neurofilaments in vitro. MAP2 bound these neurofilaments according to both SDS-PAGE and QF-DE electron microscopy of the pellets of suspensions containing L proteins and MAP2. When we added tubulin to this suspension and polymerized it in the presence of taxol, neurofilaments were crosslinked with microtubules by MAP2 crossbridges. Hence, from these 2 approaches we concluded that MAP2 is a component of crossbridges between MTs and NFs in the neuronal cytoskeleton in vivo and in vitro.


Pathology and tissue distribution of turkey coronavirus in experimentally infected chicks and turkey poults.
Twenty 1-day-old specific pathogen free chicks and 20 1-day-old commercially derived turkey poults were inoculated with a Brazilian strain of turkey coronavirus (TCoV) to study the pathogenicity and virus distribution up to 14 days post-inoculation by histopathology, immunohistochemistry, reverse transcriptase polymerase chain reaction and sequencing. At 2-14 dpi, TCoV antigens were detected in the paranasal sinus and lachrymal accessory gland (Harderian gland) of infected chicks and in the ileum, ileocaecal junction and caecum of infected poults. Lymphocytic inflammation was present in these tissues. TCoV was re-isolated from pooled tissue suspensions of nasal concha, Harderian gland and paranasal sinus from chicks, as well as from the ileum, ileocaecal junction and caecum of poults, after three consecutive passages in 28-day-old embryonated turkey eggs. Viral RNA corresponding to the spike gene region (1178-2073 genome position) was amplified from the upper respiratory tract of chickens and from the intestinal tract of poults and phylogenetic analysis confirmed the identity as TCoV. This is the first description of TCoV antigens and mRNA in upper respiratory tissues in experimentally infected chickens.


Morphine for Refractory Dyspnea in Interstitial Lung Disease: A Phase I Study (JORTC-PAL 05).
Background: Dyspnea is common in interstitial lung disease (ILD) patients and often refractory to conventional treatment. Little is known regarding the safety of systemic morphine in ILD patients. Objective: The objective of this study is to evaluate the safety of a single subcutaneous morphine injection and to determine the recommended dose of morphine for alleviating dyspnea in ILD patients. Design: We conducted a dose-escalation Phase I study for investigating the recommended dose of a single subcutaneous morphine injection to alleviate dyspnea in ILD patients. Setting/Subjects: Eligible subjects were ILD inpatients with dyspnea at rest who were refractory to conventional dyspnea treatment. The morphine doses used were 1 mg and 2 mg in cohort 1 and cohort 2, respectively. The primary endpoint was dose-limiting toxicity, which was defined as (1) respiratory depression, that is, 30% reduction of respiratory rate and 10 Torr increase of PaCO2 compared with baseline; (2) hypotension, that is, 20% reduction of systemic blood pressure compared with baseline and presentation of hypotension-related symptoms; or (3) grade 3, 4, or 5 treatment-emergent adverse events graded by Common Terminology Criteria for Adverse Events (version 4). Results: A total of six patients were enrolled, with three patients each in cohorts 1 and 2. No dose-limiting toxicities were observed; three patients experienced worsened somnolence, but no patients experienced sedation. Conclusion: We conclude that 2 mg of morphine has a tolerable safety profile in ILD patients with dyspnea, and can be tested in further clinical trials.


Central corneal thickness measurements and ultrasonographic study of the growing equine eye.
The aim of this study was to determine the central corneal thickness (CCT) of healthy equine eyes with high-frequency ultrasound (HF) and ultrasound biomicroscopy (UBM) and to measure the axial dimensions with B-mode ultrasound. Additionally, the influence of the age on the measured dimensions of the globe was analyzed. Fifty warm blood horses (mean age 23.0 ± 21.12 months) were divided into 3 age groups (group 1: 0-6 months; group 2: ≥7-30 months; and group 3: ≥31-78 months). Corneal measurements were performed with a 22-MHz and a 50-MHz transducer. Crystalline lens thickness (CLT), vitreous chamber depth (VCD), and axial globe length (AGL) were measured at a frequency of 10 MHz. Anterior chamber depth (ACD) was calculated. The eyes were measured under general anesthesia nonrelated to this study. The mean CCT of 50 eyes was 818 ± 41 μm. The mean values ± SD in mm for axial dimensions were AGL 34.7 ± 2.7, ACD 3.4 ± 0.7, CLT 10.0 ± 0.9, and VCD of 20.4 ± 1.5. A positive correlation with increasing age was found for all dimensions. No significant differences were found between the gender and the left or right eye. HF and UBM are valuable high-resolution imaging tools for CCT measurements in horses. The here-obtained values support previous findings that a positive correlation with increasing age exists for CCT, ACD, CLT, VCD, and AGL in horse.


Oxidative stress & male infertility.
The male factor is considered a major contributory factor to infertility. Apart from the conventional causes for male infertility such as varicocoele, cryptorchidism, infections, obstructive lesions, cystic fibrosis, trauma, and tumours, a new and important cause has been identified: oxidative stress. Oxidative stress is a result of the imbalance between reactive oxygen species (ROS) and antioxidants in the body. It is a powerful mechanism that can lead to sperm damage, deformity and eventually, male infertility. This review discusses the physiological need for ROS and their role in normal sperm function. It also highlights the mechanism of production and the pathophysiology of ROS in relation to the male reproductive system and enumerate the benefits of incorporating antioxidants in clinical and experimental settings.


Diversity of Contact-Dependent Growth Inhibition Systems of Pseudomonas aeruginosa
Contact-dependent growth inhibition (CDI) systems are used in bacterial competition to hinder the growth of neighboring microbes. These systems utilize a two-partner secretion mechanism to display the CdiA exoprotein at the bacterial cell surface. CdiA forms a long filamentous stalk that facilitates binding to a target cell and delivery of a C-terminal toxin (CT) domain. This CT domain is processed and delivered into the cytoplasm of a target cell upon contact. CDI systems also encode a cognate immunity protein (CdiI) that protects siblings and resistant targeted cells from intoxication by high-affinity binding to the CT. CdiA CT domains vary among strains within a species, and many alleles encode enzymatic functions that target nucleic acids. This variation is thought to help drive diversity and adaptation within a species. CdiA diversity is well studied in Escherichia coli and several other bacteria, but little is known about the extent of this diversity in Pseudomonas aeruginosa. The purpose of this review is to highlight the variability that exists in CDI systems of P. aeruginosa. We show that this diversity is apparent even among strains isolated from a single geographical region, suggesting that CDI systems play an important role in the ecology of P. aeruginosa.


Utilization of cinnamaldehyde and zinc oxide nanoparticles in a carboxymethylcellulose-based composite coating to improve the postharvest quality of cherry tomatoes.
Carboxymethylcellulose (CMC)-based film packaging enriched with plant-sourced cinnamaldehyde (CIN) and zinc oxide nanoparticles (ZnONPs) of simple synthesis were prepared in this work. Then, the physico-mechanical and barrier properties and antifungal activities of CMC-based films, including pristine CMC, CIN/CMC, ZnONPs/CMC, ZnONPs/CIN/CMC were investigated. The results indicated that the ZnONPs incorporation produced a nanocomposite film with low transparency, good flexibility and high mechanical resistance. Furthermore, the water barrier capacity and antifungal performance of CMC-based films were significantly improved with the addition of CIN. CMC-based composite film incorporating with both CIN and ZnONPs, in turn, exhibited satisfactory mechanical characterizations, excellent barrier capacities against water and oxygen molecules and anti-Aspergillus niger activity. Moreover, ZnO/CIN/CMC nanocomposite film was significantly effective at inhibiting the weight loss and firmness of cherry tomatoes and in decreasing the total acidity content of these fruits after storage. Our results suggested that the ZnO/CIN/CMC nanocomposite film packaging can improve the cherry tomatoes quality by suppressing physiological the metabolic activities of the fruits during postharvest storage period.


Raman tweezers as an alternative diagnostic tool for paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disease characterized by hemolysis of red blood cells (RBC) and venous thrombosis. The gold standard method for the diagnosis of this disease is flow cytometry. Here, we propose a combined optical tweezers and Raman spectral (Raman tweezers) approach to analyze blood samples from volunteers with or without PNH conditions. Raman spectroscopy is a well-known method for investigating a material's chemical structure and is also used in molecular analysis of biological compounds. In this study, we trap individual RBCs found in whole blood samples drawn from PNH patients and the control group. Evaluation of the Raman spectra of these cells by band component analysis and machine learning shows a significant difference between the two groups. The specificity and the sensitivity of the training performed by support vector machine (SVM) analysis were found to be 81.8% and 78.3%, respectively. This study shows that an immediate and high accuracy test result is possible for PNH disease by employing Raman tweezers and machine learning.


Functional role of hCngb3 in regulation of human cone cng channel: effect of rod monochromacy-associated mutations in hCNGB3 on channel function.
The human cone photoreceptor cyclic nucleotide-gated (CNG) channel comprises alpha- and beta-subunits, which are respectively encoded by hCNGA3 and hCNGB3. The purpose was to examine the functional role of hCNGB3 in modulation of human cone CNG channels and to characterize functional consequences of rod monochromacy-associated mutations in hCNGB3 (S435F and D633G). Macroscopic patch currents were recorded from human embryonic kidney (HEK) 293 cells expressing homomeric (hCNGA3 and hCNGB3) and heteromeric (hCNGA3/hCNGB3, hCNGA3/hCNGB3-S435F, and hCNGA3/hCNGB3-D633G) channels using inside-out patch-clamp technique. Both hCNGA3 homomeric and hCNGA3/hCNGB3 heteromeric channels were activated by cGMP, with half-maximally activating concentration (K(1/2)) of 11.1 +/- 1.0 and 26.2 +/- 1.9 micro M, respectively. The hCNGA3 channels appeared to be more sensitive to inhibition by extracellular Ca(2+) compared with hCNGA3/hCNGB3 channels, when assessed by the degree of outward rectification. Coexpression of either of rod monochromacy-associated mutants of hCNGB3 with hCNGA3 significantly reduced K(1/2) value for cGMP but little affected the sensitivity to extracellular Ca(2+), compared with wild-type heteromeric channels. The selectivity of hCNGA3, hCNGA3/hCNGB3, hCNGA3/hCNGB3-S435F, and hCNGA3/hCNGB3-D633G channels for monovalent cations were largely similar. Immunoprecipitation experiments showed association of hCNGA3 subunit with both of wild-type and mutant hCNGB3 subunits. The hCNGB3 plays an important modulatory role in the function of human cone CNG channels with respect to cGMP and extracellular Ca(2+) sensitivities. The rod monochromacy-associated S435F and D633G mutations in hCNGB3 evokes a significant increase in the apparent affinity for cGMP, which should alter cone function and thereby contribute at least partly to pathogenesis of the disease.


Wuchereria bancrofti: Unusual Presentation as Pancytopenia.
Wuchereria bancrofti is an endemic filarial nematode spread by a mosquito vector. The clinical manifestations vary from asymptomatic microfilaremeia to lymhoedema. We report two cases of microfilaria of Wuchereria bancrofti in bone marrow presenting as peripheral pancytopenia. The patient did not have any classical features of lymphoedema or elephantiasis. The diagnostic demonstration of microfilaria with unusual presentations has been done in cytology specimens from various sites. Microfilaria infection in association with peripheral blood pancytopenia has been rarely reported. An aetiopathological co-relation of these haematological manifestations of microfilaria needs further investigation.


Interdomain interface-mediated target recognition by the Scribble PDZ34 supramodule.
Tandem-arranged PDZ [PSD-95 (postsynaptic density-95), Dlg (discs large homologue) and ZO-1 (zonula occludens-1)] domains often form structural and functional supramodules with distinct target-binding properties. In the present study, we found that the two PDZ domains within the PDZ34 tandem of Scribble, a cell polarity regulator, tightly pack in a 'front-to-back' mode to form a compact supramodule. Although PDZ4 contains a distorted αB/βB pocket, the attachment of PDZ4 to PDZ3 generates an unexpected interdomain pocket that is adjacent to and integrates with the canonical αB/βB pocket of PDZ3 to form an expanded target-binding groove. The structure of the PDZ34-target peptide complex further demonstrated that the peptide binds to this expanded target-binding groove with its upstream residues anchoring into the interdomain pocket directly. Mutations of the interdomain pocket and disruptions of the PDZ34 supramodule both interfere with its target-binding capacity. Therefore, the interdomain interface between the PDZ34 supramodule is intrinsically required for its target recognition and determines its target-binding specificity. This interdomain interface-mediated specific recognition may represent a novel mode of target recognition and would broaden the target-binding versatility for PDZ supramodules. The supramodular nature and target recognition mode of the PDZ34 tandem found in the present study would also help to identify the new binding partners of Scribble and thus may direct further research on the PDZ domain-mediated assembly of Scribble polarity complexes.


The Amsler chart: absence of evidence is not evidence of absence.
Early detection of the onset or progression of macular disease is likely to become increasingly important as new treatment modalities are introduced. Current best practice involves issuing patients with an Amsler chart for daily or weekly observation with the instruction to attend for immediate assessment should any new distortion be perceived. However the sensitivity of Amsler charts in detecting macular disease can be less than 50%, implying that presentation may be delayed in over half of patients with advancing disease relying on the Amsler chart to detect progression. A likely explanation for this is the phenomenon of perceptual completion whereby regular objects are "filled-in" across the scotoma. Although alternative tests have been developed and shown to have greater sensitivity, at present no straightforward, cheap, home-based test of macular disease progression is available. The development of such a self-diagnostic tool should be a research priority.


No complexity-stability relationship in empirical ecosystems.
Understanding the mechanisms responsible for stability and persistence of ecosystems is one of the greatest challenges in ecology. Robert May showed that, contrary to intuition, complex randomly built ecosystems are less likely to be stable than simpler ones. Few attempts have been tried to test May's prediction empirically, and we still ignore what is the actual complexity-stability relationship in natural ecosystems. Here we perform a stability analysis of 116 quantitative food webs sampled worldwide. We find that classic descriptors of complexity (species richness, connectance and interaction strength) are not associated with stability in empirical food webs. Further analysis reveals that a correlation between the effects of predators on prey and those of prey on predators, combined with a high frequency of weak interactions, stabilize food web dynamics relative to the random expectation. We conclude that empirical food webs have several non-random properties contributing to the absence of a complexity-stability relationship.


Anesthesia for robotic thoracic surgery.
Robotically assisted thoracic surgery (RATS) to date is performed as a small proportion of thoracic surgical procedures and only at a few centres, but is rapidly gaining more widespread interest. In this article, we present our anesthetic perspective of planning and performing RATS and highlight specific points to consider when managing these patients.


Application of coliphage as biocontrol agent in combination with gamma irradiation to eliminate multi-drug-resistant E. coli in minimally processed vegetables.
Biofilm formation is a rising concern in the food industry. Escherichia coli (E. coli) is one of the most important food-borne pathogens that can survive in food and food-related environments and eventually produce biofilms. This study suggested that both coliphages used were successful in preventing the creation of new biofilms as well as removing existing ones. Confocal laser scanning microscopy verified these findings. According to the findings, neither coliphage survived at 37 °C, but both remained stable at 4 °C and - 20 °C for extended periods of time. The study revealed that both coliphages demonstrated a greater degree of gamma irradiation resistance when compared to E. coli. The study's results indicate that the implementation of a dual method, which incorporates gamma irradiation (1.5 kGy) and coliphage treatment, on various kinds of vegetables that were infected with E. coli, resulted in a significant reduction in bacterial count (surpassing 99.99%) following a 24-h incubation period. Combining gamma irradiation and the coliphage approach was significantly effective at lowering polysaccharide concentrations and proteins in the biofilm matrix. The results revealed that the pairing of gamma irradiation and coliphages acted in conjunction to cause disruptions in the matrix of biofilm, thereby promoting cell removal compared with either of the individual treatments. Ca+ ions strengthen the weak virion interaction with the relevant bacterial host cell receptors during the adsorption process. In conclusion, use of coliphage in combination with gamma irradiation treatment can be applied to improve fresh produce's microbial safety and enhance its storability in supermarkets.


Adherence self report assessment in solid-organ pre and post transplant recipients.
There is strong evidence that many patients with chronic diseases have difficulties to adhere to the medical regimens. As to transplantation, nonadherence to the prescribed medical regimen has repeatedly been found to predict morbidity and mortality, both in adults and in children. Many instruments are available in order to assess nonadherence to immunosuppressive medication, whereas only few are available on behavioural components. Aim of the study was to present the Adherence Schedule in Transplantation (ASiT, in its three versions: ASiT-A, Adults; ASiT-PA, Proxy Adult and ASiT-PC, Proxy Child) that evaluates the Cognitive Relational Antecedents of adherence to treatment and the Self-efficacy in disease management in patients before and after transplantation. (1) Review and construct analysis of the already existing adherence Schedules, on the basis of literature review and clinical experience; (2) Construction of the ASiT in its three versions from an adaptation of the already existing adherence Schedules; (3) Administration of the ASiT to pre and post transplant recipients followed by a semi-structured interview in order to discuss with the patients pros and cons; (4) ASiT correction on the basis of patients' comments; (5) Translation in English by a bilingual English mother tongue. On the whole 56 Schedules were administered to pre and post transplant (liver, heart, lung and kidney) recipients and their proxy and resulted to have good construct validity, were easily administered and were well accepted by patients. As to research the ASiT could cover a wide range of nonadherence sources. Within a clinical context our Schedule could be a not time consuming tool useful in favouring the communication about adherence and able to enhance patients' personal limits and resources.


Construction, characteristics, and constraints of accelerating beams based on caustic design.
Caustic methods have been proposed for wavefront design to enable light beams propagating along curved trajectories, namely accelerating beams. Here we elaborate the complete construction, remarkable characteristics, and hidden constraints of these methods. It is found that accelerating beams based on the caustic design have not only a well-known curved intensity distribution but also a linear phase distribution along the caustic proportional to the curved length, as if light field indeed moved along the caustic. Moreover, with this characteristic, further light-ray analyses are implemented to illustrate the constraints of caustic design in different cases. We expect our work will clarify some confusion on the effectiveness and applicability of caustic methods, and thus facilitate the design of accelerating beams for various applications.


Does dietary fat affect inflammatory markers in overweight and obese individuals?-a review of randomized controlled trials from 2010 to 2016.
Obesity, a major cause of death and disability, is increasing worldwide. Obesity is characterized by a chronic, low-grade inflammatory state which is suggested to play a critical role in the development of obesity-related diseases like cardiovascular diseases and type 2 diabetes. In fact, in the hours following consumption of a meal, a transient increase in inflammatory markers occurs, a response that is exaggerated in obese subjects. Dietary composition, including content of dietary fatty acids, may affect this inflammatory response both acutely and chronically, and thereby be predictive of progression of disease. The aim of the review was to summarize the literature from 2010 to 2016 regarding the effects of dietary fat intake on levels of inflammatory markers in overweight and obesity in human randomized controlled trials. We performed a literature search in MEDLINE, EMBASE, and PubMed databases. The literature search included human randomized controlled trials, both postprandial and long-term interventions, from January 2010 to September 2016. In total, 37 articles were included. Interventions with dairy products, vegetable oils, or nuts showed minor effects on inflammatory markers. The most consistent inflammatory-mediating effects were found in intervention with whole diets, which suggests that many components of the diet reduce inflammation synergistically. Furthermore, interventions with weight reduction and different fatty acids did not clearly show beneficial effects on inflammatory markers. Most interventions showed either no or minor effects of dietary fat intake on inflammatory markers in overweight and obese subjects. To progress our understanding on how diet and dietary components affect our health, mechanistic studies are required. Hence, future studies should include whole diets and characterization of obese phenotypes at a molecular level, including omics data and gut microbiota.


Synthesis of polynucleotide analogs containing a polyvinyl alcohol backbone.
Water soluble homo-base polynucleotide analogues were synthesized in which polyvinyl alcohol and partially phosphonated polyvinyl alcohol constituted the backbones,onto which were grafted uracil or adenine via 1,3-dioxane spacers formed by acetal formation with the 1,3-diol moieties in PVA. The resulting adenine-PVA polynucleotide analogs exhibited hyperchromic effects, which was not the case for the corresponding uracil compounds. Mixtures of the adenine- and aracil PVA-phosphate polynucleotide analogs in solutions exhibited characteristic S-shaped UV-absorbance vs temperature and melting curves with melting points at approximately 40 degrees C.


Fluorous membrane ion-selective electrodes for perfluorinated surfactants: trace-level detection and in situ monitoring of adsorption.
Ion-selective electrodes (ISEs) with fluorous anion-exchanger membranes for the potentiometric detection of perfluorooctanoate (PFO(-)) and perfluorooctanesulfonate (PFOS(-)) were developed. Use of an anion-exchanger membrane doped with the tetraalkylphosphonium derivative (Rf8(CH2)2)(Rf6(CH2)2)3P(+) and an optimized measurement protocol resulted in detection limits of 2.3 × 10(-9) M (1.0 ppb) for PFO(-) and 8.6 × 10(-10) M (0.43 ppb) for PFOS(-). With their higher selectivity for PFO(-) over OH(-), membranes containing the alternative anion exchanger (Rf6(CH2)3)3PN(+)P((CH2)3Rf6)3 with a bis(phosphoranylidene)ammonium group further improved the detection limit for PFO(-) to 1.7 × 10(-10) M (0.070 ppb). These values are comparable with results obtained using well-established techniques such as gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), and liquid chromatography-tandem mass spectrometry (LC-MS-MS), but the measurement with ISEs avoids lengthy sample preconcentration, can be performed in situ, and is less costly. Even when eventual spectrometric confirmation of analyte identity is required, prescreening of large numbers of samples or in situ monitoring with ISEs may be of substantial benefit. To demonstrate a real-life application of these electrodes, in situ measurements were performed of the adsorption of PFOS(-) onto Ottawa sand, which is a standard sample often used in environmental sciences. The results obtained are consistent with those from an earlier LC-MS study, validating the usefulness of these sensors for environmental studies. Moreover, PFOS(-) was successfully measured in a background of water from Carnegie Lake.


The junctions between the repetitive and the short unique sequences of the herpes simplex virus genome are determined by the polypeptide-coding regions of two spliced immediate-early mRNAs.
Immediate-early (IE) mRNAs-4 and -5 of herpes simplex virus type 2 (HSV-2) are transcribed from the IRs/TRs genome regions towards the Us region. Each of these spliced mRNAs has an untranslated leader sequence of 249 bases and a single intron of approximately 540 bases which are contained entirely within TRs/IRs sequences. The DNA sequence of the intron largely comprises tandem reiterations of three distinct short sequences. Upstream of the common 5' mRNA termini the DNA sequence contains regions of homology with the equivalent region of HSV-1. Comparison of the polypeptides encoded by these HSV-2 mRNAs with those of HSV-1 shows blocks of conserved amino acids. The locations of the first initiator ATG triplets of these two HSV-2 mRNAs suggest that the IRs/TRs regions of HSV expand, by gene conversion or by equal though non-homologous crossover, to an extent determined by the functions of the DNA sequences which are duplicated or deleted as a result of the crossover. This mechanism for expansion of repeats may apply to other herpesviruses which have a genome structure similar to that of HSV.


Multiple affinity forms of the calcitonin gene-related peptide receptor in rat cerebellum.
Binding of 125I-calcitonin gene-related peptide (125I-CGRP) to rat cerebellum membranes and the sensitivity to guanine nucleotides of binding were investigated. Cerebellum binding sites labeled by 125I-CGRP appear to be highly specific, inasmuch as CGRP inhibited binding with an IC50 of 100 pM but other peptides were inactive or much less active in displacing 125I-CGRP from these sites. 125I-CGRP binding sites in cerebellum membranes were saturable and of high affinity. Scatchard analysis of the saturation binding data revealed a homogeneous population of binding sites, with a KD of 224 +/- 28 pM and Bmax of 131 +/- 15 fmol/mg of protein. In the presence of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) (100 microM), a single population of binding sites, with a KD of 464 +/- 77 pM and Bmax of 100 +/- 14 fmol/mg of protein, was observed. The kinetics of association of 125I-CGRP with cerebellum membranes were monophasic at all ligand concentrations tested. However, the observed association rate constant (kobs) was not dependent on [125I-CGRP] in a linear fashion in either the absence or the presence of GTP gamma S (100 microM). The kinetics of dissociation of 125I-CGRP from cerebellum membranes were multiexponential, with fast and slow dissociating components having rate constants of 0.34 +/- 0.01 and 0.025 +/- 0.001 min-1, respectively. The fast dissociating component represented 60 +/- 2% of the total specific binding sites. Dissociation of 125I-CGRP from cerebellum sites was much faster in the presence of GTP gamma S (100 microM) but still exhibited dissociation from two affinity components. The rate constants for these components of dissociation were 0.67 +/- 0.03 and 0.077 +/- 0.007 min-1, with the faster dissociating component representing 66 +/- 1% of the total specific binding sites. These findings provide the first evidence that CGRP receptors exist in multiple affinity states and that cerebellum CGRP receptors are regulated by guanine nucleotides. Our results also suggest the existence of two affinity states of the CGRP-receptor-guanine nucleotide-binding protein ternary complex.


A Model for the Characterization of the Polarizability of Thin Films Independently of the Thickness of the Film.
The dielectric properties of thin films can be modified relative to the bulk material because the interaction between film and substrate influences the mobility of the atoms or molecules in the first layers. Here we show that a strong scale effect occurs in nanometer size octadecylammine thin films. This effect is attributed to the different distribution of molecules depending on the size of the film. To accurately describe this effect, we have developed a model which is a reinterpretation of the linearized Thomas-Fermi approximation. Within this model, we have been able to characterize the polarizability of thin films independently of the thickness of the film.


Prediction of clinically relevant safety signals of nephrotoxicity through plasma metabolite profiling.
Addressing safety concerns such as drug-induced kidney injury (DIKI) early in the drug pharmaceutical development process ensures both patient safety and efficient clinical development. We describe a unique adjunct to standard safety assessment wherein the metabolite profile of treated animals is compared with the MetaMap Tox metabolomics database in order to predict the potential for a wide variety of adverse events, including DIKI. To examine this approach, a study of five compounds (phenytoin, cyclosporin A, doxorubicin, captopril, and lisinopril) was initiated by the Technology Evaluation Consortium under the auspices of the Drug Safety Executive Council (DSEC). The metabolite profiles for rats treated with these compounds matched established reference patterns in the MetaMap Tox metabolomics database indicative of each compound's well-described clinical toxicities. For example, the DIKI associated with cyclosporine A and doxorubicin was correctly predicted by metabolite profiling, while no evidence for DIKI was found for phenytoin, consistent with its clinical picture. In some cases the clinical toxicity (hepatotoxicity), not generally seen in animal studies, was detected with MetaMap Tox. Thus metabolite profiling coupled with the MetaMap Tox metabolomics database offers a unique and powerful approach for augmenting safety assessment and avoiding clinical adverse events such as DIKI.


Ontogenetic changes in genetic variances of age-dependent plasticity along a latitudinal gradient.
The expression of phenotypic plasticity may differ among life stages of the same organism. Age-dependent plasticity can be important for adaptation to heterogeneous environments, but this has only recently been recognized. Whether age-dependent plasticity is a common outcome of local adaptation and whether populations harbor genetic variation in this respect remains largely unknown. To answer these questions, we estimated levels of additive genetic variation in age-dependent plasticity in six species of damselflies sampled from 18 populations along a latitudinal gradient spanning 3600 km. We reared full sib larvae at three temperatures and estimated genetic variances in the height and slope of thermal reaction norms of body size at three points in time during ontogeny using random regression. Our data show that most populations harbor genetic variation in growth rate (reaction norm height) in all ontogenetic stages, but only some populations and ontogenetic stages were found to harbor genetic variation in thermal plasticity (reaction norm slope). Genetic variances in reaction norm height differed among species, while genetic variances in reaction norm slope differed among populations. The slope of the ontogenetic trend in genetic variances of both reaction norm height and slope increased with latitude. We propose that differences in genetic variances reflect temporal and spatial variation in the strength and direction of natural selection on growth trajectories and age-dependent plasticity. Selection on age-dependent plasticity may depend on the interaction between temperature seasonality and time constraints associated with variation in life history traits such as generation length.


Light in the tunnel or just a train; impact of supply chain finance solutions on financial service providers' financial performance by mitigating financial risk.
Supply chain finance is newly emerging concept and grab attend of financial sercive providers, buyers and suppliers. This study empirically examines the impact of supply chain finance solutions (SCFS), banks financial risk on financial service providers' financial performance using panel dataset of Asian Development Bank registered countries (Pakistan, China and Bangladesh) from 2012-2021. By breaking new ground, supply chain finance solution index is developed by combining several solutions to measure its impact on financial service provider financial performance. The results show a significant impact of supply chain finance solutions on financial performance of financial service providers. Furthermore, by offering SCF solutions a bank is able to reduce its financial risk for the external parties (e.g., investors, shareholders) This research encourages financial service providers (banks) to embrace the supply chain finance solution to enhance financial performance and allows them to evaluate their supply chain finance solutions investments as a technique to mitigate financial risk.


The future of Veterans Affairs Medical Centers.
In the past four years, the Department of Veterans Affairs (VA) has experienced unprecedented changes in the ways it provides medical care, trains medical residents, and supports its clinical research program. For the most part, these changes have improved the quality and efficiency of care provided to veterans, and they have improved the chances that the VA will survive in an increasingly competitive medical market place. While the changes in priorities for training medical residents and funding clinical research have been designed to be more consistent with the overall mission of the VA, these changes have been stressful for many of the VA/medical school affiliations. Our challenge is to understand and manage these changes so that the many benefits that have derived from more than fifty years of VA/medical school affiliations can be retained.


Study of Dosage-Dependent Effects of Cytostatic Drugs Using a Fibroblast Cell Culture of the Human Nasal Mucosa.
Introduction Knowing a concentration at which cytostatic drugs are toxic for the nasal fibroblasts will enable the use cytostatic drugs in the clinical practice to prevent excessive cicatrization. Objective To determine the cytostatic concentrations of mitomycin С, doxorubicin, and 5-fluorouracil affecting nasal mucosa fibroblasts. Methods We obtained material during an endonasal dacryocystorhinostomy with the patient's informed consent. The cells were cultivated. Second- to fourth-passage cells were used in the experiments. The cells were stained for vimentin and cluster of differentiation 90 (CD90). An MTS test 3 (3-(4,5-dimethylthiazole-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium); cell viability test was performed. Results The cytostatic drugs have a toxic effect on cultivated fibroblasts of the nasal mucosa. This effect is dose-dependent. In terms of reducing the level of tissue fibrotisation in the nasal cavity, the most justified approach is to carry out an experimental study of the effect of mitomycin C, doxorubicin, and 5-fluorouracil at the concentrations of 0.25 mg/ml, 0.25 mg/ml, and 12.5 mg/ml respectively. Conclusion The authors argue that it is inappropriate to use these cytostatic drugs to conduct studies with the goal of analyzing their antifibrotic effect on the nasal mucosa at concentrations that are either lower or higher than the aforementioned ones.


Variation Between and Within Hospitals in Single Injection Caudal Local Anesthetic Dose: A Report From the Pediatric Regional Anesthesia Network.
Given that variation exists in health care utilization, expenditure, and medical practice, there is a paucity of data on variation within the practice of anesthesia. The Pediatric Regional Anesthesia Network (PRAN) data lend itself to explore whether different medical practice patterns exist and if there are nerve blocks with more local anesthetic dosing variation than others. The primary aim of this study was to quantify variation in single injection caudal block dosing, and the secondary aim was to explore possible causes for variation (eg, number of blocks performed versus geographic location). We queried the PRAN database for single injection caudal blocks in children <1 year of age. Data were analyzed for local anesthetic dose, variation within and across institutions, and possible causes. Mean dose of bupivacaine equivalents per kilogram (BE·kg) among sites ranged from 1.39 to 2.22 with an interdecile range (IDR) containing the mid 80% of all doses ranging from 0.21 to 1.48. Mean dose (BE·kg) was associated with site, age, weight, and local anesthetic used (all P < .0001). Cohen's F effect size estimate was 10 times higher for site (0.65) than for age (0.05) or weight (0.02). Variation (IDR) was not related to number of blocks done at each site (P = .23). Mean volume per kilogram was 0.9± ± 0.2 (mean ± ±standard deviation) and was more strongly associated with site (Cohen's F 0.3) than age (0.04) or weight (0.07). Wide variation in caudal local anesthetic dosing and administered volume exists. This variation is independent of the number of cases performed at each center but rather is determined by study site (ie, variation between centers) with considerable additional variation within study centers, suggesting additional variability dependent on individual practitioners. While there are legitimate reasons to vary dosing, the current approach is inconsistent and not supported by strong evidence over giving a standardized dose.


Correction.
Because a line was inadvertently omitted, the article "Reagan reforms create upheaval at NIOSH" (News and Comment, 9 Oct., p. 166) stated on p. 168 that Donald Millar, new director of the National Institute for Occupational Safety and Health (NIOSH), believes that NIOSH should be separated from the Centers for Disease Control in Atlanta. The sentence should have read, "Millar says one of the things he learned as acting director [of NIOSH] is that NIOSH should be moved to Atlanta or that its ties to CDC should be severed." Millar, who wants strengthened ties between NIOSH and CDC, favors the former alternative.


Patient preferences for first-line oral treatment for mild-to-moderate ulcerative colitis: a discrete-choice experiment.
Patients with ulcerative colitis (UC) frequently require long-term therapy to prevent relapse. Treatments such as 5-aminosalicylic acid (5-ASA [mesalazine]) are efficacious and well tolerated, but adherence to treatment is often poor. This discrete-choice experiment (DCE) was conducted to estimate differences in patient preferences for 5-ASA treatment in mild-to-moderate UC based on levels of self-reported adherence. Inclusion of patients residing in the US, UK, Germany, and Canada allowed for assessment of possible cultural differences in patient preferences. DCE attributes were determined through literature review, clinician consultation, and patient interviews. Six treatment attributes were identified: ease of swallowing, time of day, quantity, extent of flare resolution, likelihood of flare occurrence, and cost. A total of 400 patients in four countries completed the DCE and adherence (Modified Morisky Scale) surveys. Data were analyzed using generalized estimating equations to estimate patient preference and willingness to pay (WTP) by levels of self-reported adherence and country of residence. All attributes had expected polarity and were significant predictors of patient preference. Self-reported 'good' versus 'poor' adherers significantly preferred symptom control (p = 0.0108) and mucosal healing (p = 0.0190) attributes. All patients stated preference for symptom control/mucosal healing and flare risk attributes; the latter attribute was significantly preferred across all countries. Country differences in patient preference for convenience versus clinical attributes were found. Overall, patients were willing to pay £29.24 ($US46.27) per month for symptom control and mucosal healing, and an additional £78.81 ($US124.70) per month for reduction in flare risk to 10% per year (WTP costs were equalized between each country using the published 2008 purchasing power parity). Those with flares in the past year significantly preferred avoiding future flares (p < 0.0001) versus other attributes, as well as lower risk of flares (10%, likelihood ratio: 0.64-0.70). Findings indicate that self-reported adherers to UC therapy have a stronger preference for clinical benefits over other treatment attributes, suggesting that positive patient assessment of effectiveness may influence adherence. Ongoing clinician assessment of patient preferences for treatment attributes, as well as education on the importance of adherence, may help improve treatment outcomes in UC.


Absolute male factor infertility. A useful model for evaluating the efficacy of cryopreserved semen.
Cryopreserved semen generally results in diminished pregnancy rates when used for donor insemination. However, coexisting female subfecundity may significantly confound the pregnancy outcome. Therefore, in order to determine the efficacy of cryopreserved semen, 43 consecutive women with absolute male factor infertility (single women or wives of azoospermic men) were inseminated with cryopreserved spermatozoa using cervical caps. Life table analysis of the results revealed a cumulative probability of conception of 84% with a monthly fecundability rate of 19% after eight insemination cycles. These results are similar to those reported using fresh semen, suggesting that the efficacy of cryopreserved semen may be higher than previously believed.


The Toluene o-Xylene Monooxygenase Enzymatic Activity for the Biosynthesis of Aromatic Antioxidants.
Monocyclic phenols and catechols are important antioxidant compounds for the food and pharmaceutic industries; their production through biotransformation of low-added value starting compounds is of major biotechnological interest. The toluene o-xylene monooxygenase (ToMO) from Pseudomonas sp. OX1 is a bacterial multicomponent monooxygenase (BMM) that is able to hydroxylate a wide array of aromatic compounds and has already proven to be a versatile biochemical tool to produce mono- and dihydroxylated derivatives of aromatic compounds. The molecular determinants of its regioselectivity and substrate specificity have been thoroughly investigated, and a computational strategy has been developed which allows designing mutants able to hydroxylate non-natural substrates of this enzyme to obtain high-added value compounds of commercial interest. In this work, we have investigated the use of recombinant ToMO, expressed in cells of Escherichia coli strain JM109, for the biotransformation of non-natural substrates of this enzyme such as 2-phenoxyethanol, phthalan and 2-indanol to produce six hydroxylated derivatives. The hydroxylated products obtained were identified, isolated and their antioxidant potential was assessed both in vitro, using the DPPH assay, and on the rat cardiomyoblast cell line H9c2. Incubation of H9c2 cells with the hydroxylated compounds obtained from ToMO-catalyzed biotransformation induced a differential protective effect towards a mild oxidative stress induced by the presence of sodium arsenite. The results obtained confirm once again the versatility of the ToMO system for oxyfunctionalization reactions of biotechnological importance. Moreover, the hydroxylated derivatives obtained possess an interesting antioxidant potential that encourages the use of the enzyme for further functionalization reactions and their possible use as scaffolds to design novel bioactive molecules.


Cerebro-costo-mandibular syndrome.
We describe two boys with the cerebro-costo-mandibular syndrome (CCMS). Both patients presented with Pierre Robin anomaly and respiratory insufficiency and died 12 hours and 10 months after birth. The first boy had muscular hypotonia, severe micrognathia, glossoptosis, short palate, preauricular tag, paraumbilical fibroma, and a small and narrow thorax. His chest roentgenographs showed marked hypoplasia of the first to tenth rib, multiple posterior rib-gaps in the only four ossified ribs. Tracheomalacia and stenosis of the left ureter was observed during autopsy. No structural cerebral anomalies were observed. Respiratory distress necessitated a tracheostomy in the second boy. He had severe micrognathia with glossoptosis and a cleft soft palate were noted. His chest roentgenograph showed a bell-shaped, small thorax with multiple dorsal rib-gap defects. CCMS is a rare disorder often associated with Pierre Robin anomaly. Chest roentgenographs show the typical posterior rib-gap defects, which are quite variable. CCMS usually occurs as an isolated event in a family. Of 41 reported families four reports describe horizontal and two describe vertical transmission of CCMS. This might imply genetic heterogeneity with autosomal recessive and autosomal dominant inheritance. Inter- and intrafamilial expression is variable. Careful family studies are necessary before genetic counseling is given.


Accurate reconstruction of horizontal parallax-only holograms by angular spectrum and efficient zero-padding.
Accurate reconstruction of digital holograms that are large in the x direction and small in the y direction, known as horizontal parallax only digital hologram (HPO-DH), must be carried out by non-paraxial propagation approaches such as the classical angular spectrum (AS) method. However, the required space-bandwidth product (SBP) for reconstruction of HPO-DHs requires billions of pixels, which is computationally intensive. Moreover, application of zero-padding for removing aliasing components would generate an unbearable computational burden. In this work, a novel AS technique that reconstructs non-paraxial HPO-DHs with low SBP is proposed. The proposed technique first employs the multi-Fourier transform plane propagation method, which avoids the increase of size in the vertical direction of the HPO-DH to be processed. The second ingredient for field calculation is coherent superposition of vertical tiles formed from the multi-Fourier transform calculations. The described methodology enables reconstruction of HPO-DHs with the AS method and reduced SBP. Efficient managing of the SBP allows implementing zero-padding strategies in the x direction. It is shown that the padding strategies can be implemented in the frequency, space, and space-frequency domains. Hence, suppression of aliased components and increase of the spatial resolution is possible at the same time. Finally, the accuracy and utility of the developed technique is proved by both numerical simulations and experiments.


RIFM fragrance ingredient safety assessment, propyl propionate, CAS Registry Number 106-36-5.
The existing information supports the use of this material as described in this safety assessment. Propyl propionate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, phototoxicity/photoallergenicity, skin sensitization, and environmental safety. Data show that propyl propionate is not genotoxic. Data on propyl propionate provide a calculated margin of exposure (MOE) >100 for the repeated dose toxicity, reproductive toxicity, and local respiratory toxicity endpoints. Data from read-across analog pentyl propionate (CAS # 624-54-4) show that there are no safety concerns for propyl propionate for skin sensitization under the current declared levels of use. The phototoxicity/photoallergenicity endpoints were evaluated based on ultraviolet (UV) spectra; propyl propionate is not expected to be phototoxic/photoallergenic. For the hazard assessment based on the screening data, propyl propionate is not persistent, bioaccumulative, and toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards. For the risk assessment, propyl propionate was not able to be risk screened as there were no reported volumes of use for either North America or Europe in the 2015 IFRA Survey.


Pharmacokinetics of PEGylated Gold Nanoparticles: In Vitro-In Vivo Correlation.
Data suitable for assembling a physiologically-based pharmacokinetic (PBPK) model for nanoparticles (NPs) remain relatively scarce. Therefore, there is a trend in extrapolating the results of in vitro and in silico studies to in vivo nanoparticle hazard and risk assessment. To evaluate the reliability of such approach, a pharmacokinetic study was performed using the same polyethylene glycol-coated gold nanoparticles (PEG-AuNPs) in vitro and in vivo. As in vitro models, human cell lines TH1, A549, Hep G2, and 16HBE were employed. The in vivo PEG-AuNP biodistribution was assessed in rats. The internalization and exclusion of PEG-AuNPs in vitro were modeled as first-order rate processes with the partition coefficient describing the equilibrium distribution. The pharmacokinetic parameters were obtained by fitting the model to the in vitro data and subsequently used for PBPK simulation in vivo. Notable differences were observed in the internalized amount of Au in individual cell lines compared to the corresponding tissues in vivo, with the highest found for renal TH1 cells and kidneys. The main reason for these discrepancies is the absence of natural barriers in the in vitro conditions. Therefore, caution should be exercised when extrapolating in vitro data to predict the in vivo NP burden and response to exposure.


Spatiotemporal amplitude and phase retrieval of Bessel-X pulses using a Hartmann-Shack sensor.
We propose a new experimental technique, which allows for a complete characterization of ultrashort optical pulses both in space and in time. Combining the well-known Frequency-Resolved-Optical-Gating technique for the retrieval of the temporal profile of the pulse with a measurement of the near-field made with an Hartmann-Shack sensor, we are able to retrieve the spatiotemporal amplitude and phase profile of a Bessel-X pulse. By following the pulse evolution along the propagation direction we highlight the superluminal propagation of the pulse peak.


Pantothenate kinase activation restores brain coenzyme A in a mouse model of pantothenate kinase associated neurodegeneration.
Pantothenate kinase associated neurodegeneration (PKAN) is characterized by a motor disorder with combinations of dystonia, parkinsonism and spasticity, leading to premature death. PKAN is caused by mutations in the PANK2 gene that result in loss or reduction of PANK2 protein function. PANK2 is one of three kinases that initiate and regulate coenzyme A biosynthesis from vitamin B5 and the ability of BBP-671, an allosteric activator of pantothenate kinases, to enter the brain and elevate coenzyme A was investigated. The metabolic stability, protein binding and membrane permeability of BBP-671 all suggest it has the physical properties required to cross the blood brain barrier. BBP-671 was detected in plasma, liver, cerebrospinal fluid and brain following oral administration in rodents demonstrating the ability of BBP-671 to penetrate the brain. The pharmacokinetic and pharmacodynamic properties of orally administered BBP-671 evaluated in cannulated rats showed that CoA concentrations were elevated in blood, liver and brain. BBP-671 elevation of whole blood acetyl-CoA served as a peripheral pharmacodynamic marker and provided a suitable method to assess target engagement. BBP-671 treatment elevated brain coenzyme A concentrations, and improved movement and body weight in a PKAN mouse model. Thus, BBP-671 crosses the blood brain barrier to correct the brain CoA deficiency in a PKAN mouse model resulting in improved locomotion and survival, providing a preclinical foundation for the development of BBP-671 as a potential treatment for PKAN. Significance Statement The blood brain barrier represents a major hurdle for drugs targeting brain metabolism. This work describes the pharmacokinetic/pharmacodynamic properties of BBP-671, a pantothenate kinase activator. BBP-671 crosses the blood brain barrier to correct the neuron-specific CoA deficiency and improve motor function in a mouse model of pantothenate kinase associated neurodegeneration. The central role of CoA and acetyl-CoA in intermediary metabolism suggests that pantothenate kinase activators may be useful in modifying neurological metabolic disorders.


Massive dispersal of Coxiella burnetii among cattle across the United States.
Q-fever is an underreported disease caused by the bacterium Coxiella burnetii, which is highly infectious and has the ability to disperse great distances. It is a completely clonal pathogen with low genetic diversity and requires whole-genome analysis to identify discriminating features among closely related isolates. C. burnetii, and in particular one genotype (ST20), is commonly found in cow's milk across the entire dairy industry of the USA. This single genotype dominance is suggestive of host-specific adaptation, rapid dispersal and persistence within cattle. We used a comparative genomic approach to identify SNPs for high-resolution and high-throughput genotyping assays to better describe the dispersal of ST20 across the USA. We genotyped 507 ST20 cow milk samples and discovered three subgenotypes, all of which were present across the entire country and over the complete time period studied. Only one of these sub-genotypes was observed in a single dairy herd. The temporal and geographic distribution of these sub-genotypes is consistent with a model of large-scale, rapid, frequent and continuous dissemination on a continental scale. The distribution of subgenotypes is not consistent with wind-based dispersal alone, and it is likely that animal husbandry and transportation practices, including pooling of milk from multiple herds, have also shaped the patterns. On the scale of an entire country, there appear to be few barriers to rapid, frequent and large-scale dissemination of the ST20 subgenotypes.


Investigating a crow die-off in January-February 2011 during the introduction of a new clade of highly pathogenic avian influenza virus H5N1 into Bangladesh.
We investigated unusual crow mortality in Bangladesh during January-February 2011 at two sites. Crows of two species, Corvus splendens and C. macrorhynchos, were found sick and dead during the outbreaks. In selected crow roosts, morbidity was ~1 % and mortality was ~4 % during the investigation. Highly pathogenic avian influenza virus H5N1 clade 220.127.116.11 was isolated from dead crows. All isolates were closely related to A/duck/India/02CA10/2011 (H5N1) with 99.8 % and A/crow/Bangladesh/11rs1984-15/2011 (H5N1) virus with 99 % nucleotide sequence identity in their HA genes. The phylogenetic cluster of Bangladesh viruses suggested a common ancestor with viruses found in poultry from India, Myanmar and Nepal. Histopathological changes and immunohistochemistry staining in brain, pancreas, liver, heart, kidney, bursa of Fabricius, rectum, and cloaca were consistent with influenza virus infection. Through our limited investigation in domesticated birds near the crow roosts, we did not identify any samples that tested positive for influenza virus A/H5N1. However, environmental samples collected from live-bird markets near an outbreak site during the month of the outbreaks tested very weakly positive for influenza virus A/H5N1 in clade 220.127.116.11-specific rRT-PCR. Continuation of surveillance in wild and domestic birds may identify evolution of new avian influenza virus and associated public-health risks.


Genetic disruption of aurora B uncovers an essential role for aurora C during early mammalian development.
Mitosis is controlled by multiple kinases that drive cell cycle progression and prevent chromosome mis-segregation. Aurora kinase B interacts with survivin, borealin and incenp to form the chromosomal passenger complex (CPC), which is involved in the regulation of microtubule-kinetochore attachments and cytokinesis. Whereas genetic ablation of survivin, borealin or incenp results in early lethality at the morula stage, we show here that aurora B is dispensable for CPC function during early cell divisions and aurora B-null embryos are normally implanted. This is due to a crucial function of aurora C during these early embryonic cycles. Expression of aurora C decreases during late blastocyst stages resulting in post-implantation defects in aurora B-null embryos. These defects correlate with abundant prometaphase figures and apoptotic cell death of the aurora B-deficient inner cell mass. Conditional deletion of aurora B in somatic cells that do not express aurora C results in chromosomal misalignment and lack of chromosome segregation. Re-expression of wild-type, but not kinase-dead, aurora C rescues this defect, suggesting functional overlap between these two kinases. Finally, aurora B-null cells partially arrest in the presence of nocodazole, suggesting that this kinase is not essential for the spindle assembly checkpoint.


FTO and MC4R polymorphisms, and selected pre-, peri- and postnatal factors as determinants of body mass index and fatness in children: a thorough analysis of the associations.
Overweight and obesity among children have become significant global health concerns. Previous studies have highlighted the potential role of genetic factors, particularly polymorphisms in the FTO and MC4R genes, as well as environmental factors in the development of childhood obesity. This study aimed to investigate the relationships between genetic, socioeconomic and perinatal factors, adverse childhood events (ACEs), and lifestyle, and their impact on overweight, obesity and body composition parameters in children. Additionally, we explored potential interactions between genetic factors and ACEs. Four hundred fifty-six children aged 6-12 years participated in our study. Information on the socioeconomic status, perinatal factors, ACEs and lifestyle of the children was collected with a questionnaire completed by their parents/guardians. We examined the children's body weight and conducted an electrical bioimpedance analysis. Overweight and obesity were diagnosed based on the International Obesity Task Force and McCarthy criteria. We genotyped two selected polymorphisms in the FTO and MC4R genes using the TaqMan SNP allelic discrimination method. Higher BMI (Body Mass Index) z scores were related to higher paternal BMI and lower maternal age at the child's birth. Higher FMI (Fat Mass Index) z scores were associated with higher paternal BMI, increased gestational weight, lower maternal education and the presence of the FTO risk allele. Higher FatM (fat mass in kg) z scores were linked to lower maternal education, lower maternal age at the child's birth, higher maternal body weight gain, paternal BMI and the presence of the FTO risk allele. Moreover, interaction effects were observed on BMI z scores between ACE and FTO AA, and on FMI z scores and FatM z scored between ACE and MC4R CC. The contribution of environmental factors is more strongly related to changes in body composition than genetic ones. Additionally, the presence of the risk allele combined with unfavourable environmental factors like ACEs leads to visible interaction effects, resulting in increased BMI z scores and FMI z scores in children.


Porphyrins fused with strongly electron-donating 1,3-dithiol-2-ylidene moieties: redox control by metal cation complexation and anion binding.
A new class of redox-active free base and metalloporphyrins fused with the 1,3-dithiol-2-ylidene subunits present in tetrathiafulvalene, termed MTTFP (M = H2, Cu, Ni, Zn), have been prepared and characterized. The strong electron-donating properties of MTTFP were probed by electrochemical measurement and demonstrated that oxidation potentials can be tuned by metalation of the free base form, H2TTFP. X-ray crystal structures of H2TTFP, ZnTTFP, and CuTTFP revealed that a severe saddle-shape distortion was observed with the dithiole rings bent out of the plane toward one another in the neutral form. In contrast, the structure of the two-electron oxidized species (CuTFFP(2+)) is planar, corresponding to a change from a nonaromatic to aromatic structure upon oxidation. A relatively large two-photon absorption (TPA) cross-section value of H2TTFP(2+) (1200 GM) was obtained for the free base compound, a value that is much higher than those typically seen for porphyrins (<100 GM). Augmented TPA values for the metal complexes were also seen. The strong electron-donating ability of ZnTTFP was further enhanced by binding of Cl(-) and Br(-) as revealed by thermal electron-transfer between ZnTTFP and Li(+)-encapsulated C60 (Li(+)@C60) in benzonitrile, which was "switched on" by the addition of either Cl(-) or Br(-) (as the tetrabutylammonium salts). The X-ray crystal structure of Cl(-)-bound ZnTTFP was determined and provided support for the strong binding between the Cl(-) anion and the Zn(2+) cation present in ZnTTFP.


EMS in the pueblos.
Imagine creating a movie by excerpting scenes from "Dances With Wolves," splicing it with footage from "Code 3" or "Emergency Response" and then flavoring the script with the mystery of a Tony Hillerman novel. A film producer would probably find it quite difficult to choreograph a finished product from such a compilation of material. To hundreds of Native American EMS providers, however, such a movie is played out every day in Indian country. And with this movie come some real-life problems, including trauma, which is the number-one cause of premature death among Native Americans. But a high trauma rate is just one of the challenges facing tribal EMS responders. There's also prolonged response and transport, the problems involved in maintaining the unique culture and standard of care, the challenges of tribal EMS administration and EMS education of Native American students, and the unsure future of Native American EMS. Beyond that, there's the fact that EMS is a s unique to each Indian reservation as are the cultures of the native peoples who reside on these lands. Yet while no two systems are alike, most tribal EMS providers face similar challenges.