Isolation and characterization of pyrene and benzo[a]pyrene-degrading Klebsiella pneumonia PL1 and its potential use in bioremediation.
Polycyclic aromatic hydrocarbons (PAHs), which are hard to degrade, are the main pollutants in the environment. Degradation of PAHs in the environment is becoming more necessary and urgent. In the current study, strain PL1 with degradation capability of pyrene (PYR) and benzo[a]pyrene (BaP) was isolated from soil and identified as Klebsiella pneumoniae by morphological and physiological characteristics as well as 16S rDNA sequence. With the presence of 20 mg L⁻¹ PYR and 10 mg L⁻¹ BaP in solution, the strain PL1 could degrade 63.4 % of PYR and 55.8 % of BaP in 10 days, respectively. The order of biodegradation of strain PL1 was pH 7.0 > pH 8.0 > pH 10.0 > pH 6.0 > pH 5.0. Strain PL1 degradation ability varied in different soil. The half-life of PYR in soil was respectively 16.9, 24.9, and 88.9 days in paddy soil, red soil, and fluvo-aquic soil by PL1 degradation; however, the half-lives of BaP were respectively 9.5, 9.5, and 34.0 days in paddy soil, red soil, and fluvo-aquic soil by PL1 degradation. The results demonstrate that the degradation capability on PYR and BaP by PL1 in paddy soil was relatively good, and K. pneumoniae PL1 was the new degradation bacterium of PYR and BaP. K. pneumoniae PL1 has potential application in PAH bioremediation.


Microneurosurgical training in the anatomical specimen: A structured plan for endoscopic and microsurgical skull base training during the residency.
The development of a high level of competence and technical proficiency is one of the main objectives of any neurosurgical training program. Due to many factors, this progressive skill development can be complex during the residency. Despite its high cost and infrastructure requirements, there is renewed interest regarding the role of anatomy labs. The study and dissection of the human cadaver has been the environment where many surgeons have developed the necessary skills for microneurosurgery. We propose a structured endoscopic and microsurgical training dissection program to enable residents to maximize the benefits of their training in the lab. During the months of September, October and November 2021, a stay was done at the Microneurosurgery and Skull Base Laboratory of the Miguel Hernández University of Alicante. A total of 2 specimens were used. The first specimen underwent a first endoscopic endonasal dissection phase. After completing the endonasal part, a set of incisions were made to perform the transcranial part. In the second specimen, the transcranial part was performed first, leaving the endonasal endoscopic work for the last phase. The results of the dissection program are presented. During the endonasal endoscopic phase, the transsphenoidal approach to the sella was simulated while focusing on the extended approaches in the sagittal plane. During the transcranial phase, right and left anterolateral approaches, a left anterior transcallosal interhemispheric approach, a left transcondylar posterolateral approach and a combined right lateral approach were performed. The structured dissection of the specimen allowed both endonasal endoscopic and transcranial microsurgical training in the same specimen. This design facilitated the realization of the core skull base approaches in the same specimen. According to our initial experience, we believe that developing common dissection programs is a powerful tool to maximize the results of our residents' laboratory training.


Renin and aldosterone are not associated with vulnerable plaque characteristics in patients with carotid artery disease.
The renin-angiotensin-aldosterone system is increasingly being recognized to play an important role in the development and clinical course of cardiovascular diseases. Renin-angiotensin-aldosterone system activation is associated with clinical outcome in various populations of cardiovascular patients, such as patients with coronary artery, peripheral artery, and cerebrovascular disease. In this study, we investigated the associations between plasma renin and aldosterone concentrations and atherosclerotic plaque characteristics and secondary vascular events in patients undergoing carotid endarterectomy. Baseline plasma renin and aldosterone concentrations from 506 subjects undergoing carotid endarterectomy (mean age, 67 ± 9 years; 65% male) were correlated with histopathologic characteristics and inflammatory protein concentrations of the excised atherosclerotic plaque. Ordinal logistic regression (for ordinal outcome parameters) or linear regression (for linear outcome) analysis did not show a statistically significant relationship between plasma renin or aldosterone concentrations and plaque fat, thrombus, calcifications, collagen, smooth muscle cells, or macrophage content. Neither could any association be found with intraplaque inflammatory mediators. During a median follow-up of 3 years, 102 (20%) patients experienced a major secondary vascular event (composite of stroke, myocardial infarction, leg amputation, vascular death, or coronary revascularization or peripheral intervention). In multivariable Cox regression analysis, including both renin and aldosterone, baseline renin concentrations were associated with the occurrence of secondary events. In patients with established atherosclerotic disease undergoing carotid endarterectomy, plasma renin and aldosterone concentrations were not associated with atherosclerotic plaque characteristics. Plasma renin concentration was positively associated with the occurrence of major secondary vascular events.


Failure of manganese to protect from Shiga toxin.
Shiga toxin (Stx), the main virulence factor of Shiga toxin producing Escherichia coli, is a major public health threat, causing hemorrhagic colitis and hemolytic uremic syndrome. Currently, there are no approved therapeutics for these infections; however manganese has been reported to provide protection from the Stx1 variant isolated from Shigella dysenteriae (Stx1-S) both in vitro and in vivo. We investigated the efficacy of manganese protection from Stx1-S and the more potent Stx2a isoform, using experimental systems well-established for studying Stx: in vitro responses of Vero monkey kidney cells, and in vivo toxicity to CD-1 outbred mice. Manganese treatment at the reported therapeutic concentration was toxic to Vero cells in culture and to CD-1 mice. At lower manganese concentrations that were better tolerated, we observed no protection from Stx1-S or Stx2a toxicity. The ability of manganese to prevent the effects of Stx may be particular to certain cell lines, mouse strains, or may only be manifested at high, potentially toxic manganese concentrations.


Does COVID-19 infection increase the risk of pressure injury in critically ill patients?: A narrative review.
Patients with severe COVID-19 may have endothelial dysfunction and a hypercoagulable state that can cause skin damage. In the presence of external pressure on the tissues, the local inflammatory process regulated by inflammatory cytokines can increase and prolong itself, contributing to the formation of pressure injury (PI). PI is defined as localized damage to the skin or underlying tissues. It usually occurs as a result of intense and/or prolonged pressure in combination with shear. The aim of the study is to perform a narrative review on the physiological evidence of increased risk in the development of PI in critically ill patients with COVID-19.In patients with severe COVID-19 a pattern of tissue damage consistent with complement-mediated microvascular injury was found in the lungs and skin of critically ill COVID-19 patients, suggesting sustained systemic activation of complement pathways. Theoretically, the same thrombogenic vascular changes related to COVID-19 that occur in the skin also occur in the underlying tissues, making patients less tolerant to the harmful effects of pressure and shear. Unlike the syndromes typical of acute respiratory illnesses and other pathologies that commonly lead to intensive care unit admission, COVID-19 and systemic viral spread show that local and systemic factors overlap. This fact may be justified by current epidemiological data showing that the prevalence of PI among intensive care unit patients with COVID-19 was 3 times higher than in those without COVID-19. This narrative review presents physiological evidence to suggesting an increased risk of developing PI in critically ill patients with COVID-19.


Evaluation of sirtuin role in neuroprotection of retinal ganglion cells in hypoxia.
PURPOSE. Hypoxia-induced apoptosis is responsible for reduced retinal ganglion cell (RGC) viability in a variety of chronic ocular disorders. Sirtuin 1 (SIRT1) plays an important role in preserving cell viability during hypoxia. We investigated the role of SIRT1 in sustaining RGC viability in an in vitro model of hypoxia. METHODS. Staurosphorine-differentiated RGCs (RGC-5) received varying hypoxic concentrations (100-500 μM) of cobalt chloride (CoCl2) for 24 hours. Hypoxia-induced cell viability was assessed by WST-1 assay. The role of SIRT1 in promoting viability was determined indirectly via sirtinol (SIRT1 inhibitor). Hypoxia-induced apoptosis was evaluated by measuring stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) and caspase 3 activity. Vascular endothelial growth factor (VEGF) was measured to ascertain the influence of SIRT1. RESULTS. CoCl2 concentrations greater than 100 μM resulted in significantly reduced RGC viability (P=0.01). CoCl2 treatment increased SIRT1 levels significantly (P<0.01): 100 (6.5-fold), 200 (6-fold), 300 (3.5-fold), and 400 μM (4.5-fold). Phosphorylated SAPK/JNK increased 36-fold (200 μM CoCl2 concentration), then plateaued at the 300- (25-fold) and 400-μM (27.8-fold) CoCl2 concentrations (P<0.01). CoCl2 and sirtinol treatment increased Caspase 3 activity (P<0.05). VEGF release was significantly higher than control at the 100-μM CoCl2 concentrations (P<0.01). Sirtinol reduced RGC viability, SIRT1 levels, and VEGF release (P<0.01) while having greater effect on SAPK/JNK phosphorylation. CONCLUSIONS. SIRT1 significantly influences RGC viability. Sirtinol's effect reflects the interaction SIRT1 has with apoptotic signaling proteins. This investigation demonstrated SIRT1 importance in forestalling the effects of hypoxia-induced apoptosis.


Daidzein and 17 beta-estradiol enhance nitric oxide synthase activity associated with an increase in calmodulin and a decrease in caveolin-1.
Isoflavones, such as daidzein, are proposed to possess vasculoprotective properties, perhaps through a mechanism similar to estrogen. Our experiments aimed to test the hypothesis that daidzein and 17 beta-estradiol enhance endothelium-dependent relaxation through an increase in NO synthesis due to an increase in activity or expression of endothelial nitric oxide synthase (eNOS). Male rats were treated with daidzein (0.2 mg/kg per day sc), 17 beta-estradiol (0.1 mg/kg per day sc), or vehicle for 7 days and reactivity of isolated aortic rings was then determined. ACh-induced relaxation was significantly enhanced in aortic rings from rats treated with daidzein or 17 beta-estradiol but the relaxant responses to the endothelium-independent dilators sodium nitroprusside or isoprenaline were not different. Nitrite production and the level of cGMP were significantly greater in aortae from daidzein and 17 beta-estradiol compared with vehicle-treated rats. Daidzein and 17 beta-estradiol did not alter eNOS protein in endothelium-intact aortae but reduced expression of caveolin-1 and increased expression of calmodulin, changes that would account for an increase in eNOS activity. There were no differences between groups in the expression of calmodulin and caveolin-1 in arteries when the endothelium was removed. Daidzein or 17 beta-estradiol treatment selectively enhances endothelium-dependent relaxation in male rats through an increase in eNOS activity. The increase in eNOS activity is associated with a decreased expression of caveolin-1 and an increased expression of calmodulin in endothelial cells.


Survival Models on Unobserved Heterogeneity and their Applications in Analyzing Large-scale Survey Data.
In survival analysis, researchers often encounter multivariate survival time data, in which failure times are correlated even in the presence of model covariates. It is argued that because observations are clustered by unobserved heterogeneity, the application of standard survival models can result in biased parameter estimates and erroneous model-based predictions. In this article, the author describes and compares four methods handling unobserved heterogeneity in survival analysis: the Andersen-Gill approach, the robust sandwich variance estimator, the hazard model with individual frailty, and the retransformation method. An empirical analysis provides strong evidence that in the presence of strong unobserved heterogeneity, the application of a standard survival model can yield equally robust parameter estimates and the likelihood ratio statistic as does a corresponding model adding an additional parameter for random effects. When predicting the survival function, however, a standard model on multivariate survival time data can result in serious prediction bias. The retransformation method is effective to derive an adjustment factor for correctly predicting the survival function.


Long-term pain relief produced by intrathecal morphine infusion in 53 patients.
The present report details the characteristics of the analgesic effects of morphine administered chronically by infusion pumps implanted in 53 patients suffering from terminal metastatic disease. The median postimplant survival time in these patients was 4 months. Patients (mean age 58 years) were characterized according to the duration of pain before pump implantation (mean 16 months), prior consumption of systemic opioids (mean one to six daily analgesic equivalents of morphine), and their response to a trial intrathecal dose of morphine (1 to 2 mg). The median infusion dose at 2 weeks was 3.8 mg/day. The analgesic index, calculated as (quality of pain relief x duration of pain relief in hours)/morphine dose in mg, that was observed after the trial dose of morphine was determined for each patient. A close correlation was observed between the acute (2-week) infusion dose necessary to produce pain relief and the analgesic index such that the infusion dose = -8.0 x log (analgesic index) + 17.1. By 16 weeks, the mean spinal morphine dose for the group had increased by a factor of about 2.5; however, significant variation in the dose incrementation was documented. The maximum increase was observed in patients with a low analgesic index, and this rapid incrementation was usually correlated with an unsatisfactory overall outcome. Evidence that long-term infusion continues to yield analgesia was evidenced in six cases where there was an unanticipated loss of drug infusion and a corresponding increase in parenteral narcotic consumption. These data indicate the long-term efficacy and safety of spinal opioid infusion in patients with terminal cancer, and emphasize the advantage of assessing the sensitivity of the patient to spinal opioids by a standardized trial injection prior to pump placement as a prognostic indication of outcome.


Prosocial behaviors during school activities among child survivors after the 2011 earthquake and Tsunami in Japan: a retrospective observational study.
The 2011 Japan massive tsunami traumatized many children. The aim of this study was to assess changes in strengths and difficulties experienced in home and school by among surviving children after the 2011 tsunami, in comparison with published normal Japanese data. In November 2012 (20 months after the disaster) and September 2013 (30 months after the disaster), the Strengths and Difficulties Questionnaire (SDQ), a questionnaire on children's strengths and difficulties in home and school activities, were distributed to 12,193 and 11,819 children, respectively. An effective response of children 20 months and 30 month after the disaster was obtained in 10,597 children (86.9%), and 10,812 children (91.4%), respectively. The SDQ scores evaluated by parents and teachers were compared with published normal Japanese SDQ scores. The SDQ scores (emotional problems, conduct problems, hyperactivity/inattention, peer relationship problems, and total difficulty score) evaluated by parents of children in the 4th to 9th grade who were evaluated after 30 and 20 months were significantly high compared with the published normal data of children without traumatic experiences (all P<0.001). The SDQ scores (prosocial behavior) evaluated by teachers of children in the 4th to 9th grade who were evaluated after 30 and 20 months were significantly low compared with the published normal data of children without traumatic experiences (all P<0.001). This study showed that the experience of the disaster affected those children with prosocial behaviors towards teachers and friends at school. However, no significant changes (in their prosocial attitude) had been seen at home, where they continued to keep their respect and caring feelings for parents. These results indicate that for accurate diagnosis, clinicians should not only evaluate these children's daily activities at home but also try to objectively assess their daily activities at school.


Cervical microbiome is altered in cervical intraepithelial neoplasia after loop electrosurgical excision procedure in china.
Although human papillomavirus (HPV) infection is a major cause leading to the development of cervical intraepithelial neoplasia (CIN), the relationship between genital microbiome and HPV persistence/clearance is not well established. Loop electrosurgical excision procedure (LEEP) is one of standard treatments of CIN 2/3 globally, yet little is known about how the LEEP influence genital microbiota. We conducted a prospective study of 26 patients with CIN2/3 who underwent analysis of cervical microbiome before and after 3 months of LEEP treatment. Cervical swabs were collected, and microbiomes were analyzed by 16S ribosomal RNA gene sequencing. A decrease of cervical microbial diversity was observed after 3 months of LEEP treatment. Notably, a significant shift from community type of a Prevotella-containing and lack of a consistent dominant species to lactobacillus iners dominated microbiome correlated with LEEP. Particularly, Leptotrichia and clostridium were further decreased after LEEP treatment (P = 0.049 and P = 0.002, respectively). Our results suggest that the cervical microbiome is altered after LEEP treatment in patients with CIN2/3. Further studies with larger sample sizes are needed to validate these findings.


Mechanistic models enable the rational use of in vitro drug-target binding kinetics for better drug effects in patients.
Drug-target binding kinetics are major determinants of the time course of drug action for several drugs, as clearly described for the irreversible binders omeprazole and aspirin. This supports the increasing interest to incorporate newly developed high-throughput assays for drug-target binding kinetics in drug discovery. A meaningful application of in vitro drug-target binding kinetics in drug discovery requires insight into the relation between in vivo drug effect and in vitro measured drug-target binding kinetics. In this review, the authors discuss both the relation between in vitro and in vivo measured binding kinetics and the relation between in vivo binding kinetics, target occupancy and effect profiles. More scientific evidence is required for the rational selection and development of drug-candidates on the basis of in vitro estimates of drug-target binding kinetics. To elucidate the value of in vitro binding kinetics measurements, it is necessary to obtain information on system-specific properties which influence the kinetics of target occupancy and drug effect. Mathematical integration of this information enables the identification of drug-specific properties which lead to optimal target occupancy and drug effect in patients.


Association of caregiver health beliefs and parenting stress with medication adherence in preschoolers with asthma.
This study examined the associations of parenting stress and caregiver negative health beliefs with medication adherence in a sample of preschool-aged children with asthma. Participants included 43 caregivers and their child with asthma, aged 2-5 years, who were prescribed a preventative or controller medication. Assessment included parent report of parenting stress associated with parental characteristics and difficult child behavior, health-related beliefs, and adherence to inhaled corticosteroids (ICS). Pharmacy refill records were reviewed to measure medication adherence objectively. Statistical analyses included paired samples t-test and Pearson's correlation coefficients. Parent-report and objective measures of adherence to ICS were significantly different, with parents reporting significantly higher rates of medication adherence (p < 0.001). Also, increased caregiver negative health beliefs, but not parenting stress, were significantly and negatively associated with an objective measure of ICS adherence (p < 0.01). Because the findings for parenting stress are not consistent with that of the literature on school-aged children with asthma, our results may suggest that preschool children are a unique developmental group. A review of endorsed health beliefs also highlights strengths and concerns for caregivers of these very young children. Overall, our study findings underscore the importance of caregivers' health-related beliefs in promoting adherence to ICS in young children with asthma.


The vestibular symptomatology of Machado-Joseph Disease.
Machado Joseph Disease (MJD) is an autosomal dominant neurodegenerative disease. In previous studies, we described significant bilateral horizontal Vestibulo-Ocular Reflex (VOR) deficit within this population without any reference to the presence of vestibular symptomatology. To evaluate whether, beyond cerebellar ataxia complaints, MJD patients have typical vestibular symptomatology corresponding to the accepted diagnostic criteria of Bilateral Vestibulopathy (BVP) according to the definition of the International Barany Society of Neuro-Otology. Twenty-one MJD, 12 clinically stable chronic Unilateral Vestibulopathy (UVP), 15 clinically stable chronic BVP, and 22 healthy Controls underwent the video Head Impulse Test (vHIT) evaluating VOR gain and filled out the following questionnaires related to vestibular symptomatology: The Dizziness Handicap Inventory (DHI), the Activities-specific Balance Confidence Scale (ABC), the Vertigo Visual Scale (VVS) and the Beck Anxiety Inventory (BAI). The MJD group demonstrated significant bilateral vestibular impairment with horizontal gain less than 0.6 in 71% of patients (0.54±0.17). Similar to UVP and BVP, MJD patients reported a significantly higher level of symptoms than Controls in the DHI, ABC, VVS, and BAI questionnaires. MJD demonstrated significant VOR impairment and clinical symptoms typical of BVP. We suggest that in a future version of the International Classification of Vestibular Disorders (ICVD), MJD should be categorized under a separate section of central vestibulopathy with the heading of bilateral vestibulopathy. The present findings are of importance regarding the clinical diagnosis process and possible treatment based on vestibular rehabilitation.


Artificial microRNA-mediated knockdown of pyruvate formate lyase (PFL1) provides evidence for an active 3-hydroxybutyrate production pathway in the green alga Chlamydomonas reinhardtii.
Artificial microRNA technology was investigated as a means of down regulating metabolic pathways in the green alga Chlamydomonas reinhardtii, targeting pyruvate formate lyase (PFL1), which catalyzes the conversion of pyruvate to acetyl-CoA and formate during anoxic conditions. Two transformants with an 80-90% reduction in target protein and mRNA levels were identified. Nuclear magnetic resonance spectroscopy confirmed a substantial decrease in the production of formate in the knockdown lines during dark anoxic conditions and a re-routing of metabolism leading to enhanced production of ethanol and lactate. Under microaerobic conditions in the light, induced by sulphur-deprivation, knock-down of PFL1 resulted in reduced formate and ethanol production, increased net consumption of acetate and the excretion of lactate but no increase in the production of hydrogen. In addition the production of 3-hydroxybutyrate was identified in knock-down line cultures during the transition between microaerobic and anoxic conditions. Overall our results indicate that microRNA knock-down is a useful tool to manipulate anaerobic metabolism in C. reinhardtii.


Are social workers required to engage in participatory practices? An analysis of job descriptions.
The participatory approach is depicted as inherent to the social work profession in social workers' theoretical literature, codes of ethics and practical discourse. The current study examined whether, and to what degree, social workers in Israel are indeed formally required to engage in participatory practices in the job descriptions of diverse social care services. A quantitative content analysis of 78 formal social workers' job descriptions revealed this requirement to be very limited. Most participatory actions required of social workers represented low-level service-user participation, pertained only to the initial and concluding stages of intervention, and referred to professional domains in which service-user participation is already required by law. A greater emphasis on participatory practice was found in job descriptions in the health field and those relating to community social workers. Various interpretations and implications of these findings are discussed.


[Target volume for adjuvant radiotherapy after prostatectomy].
Although radical prostatectomy is an effective treatment for clinically localized prostate cancer, it fails in up to 20 to 40% of the cases. Local failure represents one of the possible patterns of relapse and is announced by detectable serum PSA levels. Patients at high risk for local relapse have extraprostatic disease, positive surgical margins or seminal vesicles infiltration at pathology. Three recently published phase III randomized clinical trials have clearly shown that, for these patients, immediate adjuvant irradiation reduces the risk of progression. For patients undergoing postoperative irradiation the standardisation of the target volume definition and delineation is required because no general consensus exists on "prostate bed" definition. The Genito-Urinary Working Party of the Radiation Oncology Group of the European Organization for the Research and treatment of cancer (ROG-EORTC) has developed a set of Guidelines to assist radiotherapists in the contouring of target volumes for postoperative irradiation: a consensus has been reached on a set of recommendations that are proposed to the radiation oncologist community. Emphasis has been put on the optimal cooperation between the surgeon, the pathologist and the radiotherapist in the frame of a multidisciplinary approach. Data on the presence and on the localization of extraprostaic extension and on positive surgical margins must be used. Placement of metallic clips in the tumor bed is of great help in localizing fixed anatomical sites as the anastomosis. The goal of such a document is to reduce inter-observer variability in target delineation in the framework of future clinical trials.


[UPLC Fingerprints of Euphorbiae Ebracteolatae Radix and Its Vinegar-Processed Slices].
To compare the qualities of Euphorbiae Ebracteolatae Radix from different original plants and its vinegar-processed slices. UPLC fingerprints of Euphoria ebracteolata, Euphoria fischeriana and vinegar-processed slices of Euphoria ebracteolata were established on a Waters ODS column (100 mm x 2.1 mm, 1.8 μm), using jolkinolide B as the reference substance. A mixture of acetonitrile and water was used as the mobile phase in a gradient elution mode, with the flow rate of 0.4 mL/min, the column temperature at 30 degrees C and the UV detector at 210 nm. The UPLC analysis time was no more than 20 min. The qualities of Euphoria ebracteolata and its vinegar-processed slices were stable with similarities over 0.90, respectively. However, the fingerprints of the slices and their original crude drugs exhibited obvious difference on both number and the relative peak area of the common peaks. Meanwhile the similarities of the fingerprints of Euphoria fischeriana and Euphoria ebracteolata were less than 0.72, indicating significant difference of these two plants. The developed method is available for quality control of Euphorbiae Ebracteolatae Radix and mechanism exploration of its vinegar-processed slices.


Association of chronic kidney disease and coronary artery disease in 1,010 consecutive patients undergoing coronary angiography.
Patients with chronic kidney disease (CKD) are more likely to have complications due to cardiovascular disease (CVD). This study was performed to investigate the prevalence of chronic kidney disease (CKD) and the relation of CKD and number of stenosed coronary vessels in patients who had undergone coronary angiography with suspected coronary artery disease (CAD). The data of 1,010 consecutive patients who underwent coronary angiography for suspected CAD in Zhongda Hospital were analyzed. Estimated glomerular filtration rate (eGFR) was calculated with the abbreviated Modified Diet in Renal Disease (MDRD) Study equation. CKD was defined as presence of eGFR <60 ml/min per 1.73 m(2) and/or proteinuria. Luminal narrowing with at least 1 lesion =50% in the main branches of the coronary artery was considered as CAD. The number of stenotic arteries was recorded (1- to 3-vessel disease [VD]). A significant stenosis in the left main trunk was scored as 2-VD. Patients with CAD had a significantly higher prevalence of CKD compared with patients without CAD (18.8% vs. 5.4%, p<0.001). CKD was significantly associated with CAD, 2-VD and 3-VD, versus without CAD (0-VD) (odds ratio [OR] = 2.163; 95% confidence interval [95% CI], 1.296-3.611; OR=2.478; 95% CI, 1.288-4.766; OR=2.504; 95% CI, 1.271-4.933; respectively) after adjustment for covariates. There was a higher prevalence of CKD in patients with CAD diagnosed by coronary angiography, and it increased with the number of stenosed coronary vessels. CKD is a critical risk factor for CAD, especially 3-VD.


Challenges in Obtaining Estimates of the Risk of Tuberculosis Infection During Overseas Deployment.
Estimates of the risk of tuberculosis (TB) infection resulting from overseas deployment among U.S. military service members have varied widely, and have been plagued by methodological problems. The purpose of this study was to estimate the incidence of TB infection in the U.S. military resulting from deployment. Three populations were examined: 1) a unit of 2,228 soldiers redeploying from Iraq in 2008, 2) a cohort of 1,978 soldiers followed up over 5 years after basic training at Fort Jackson in 2009, and 3) 6,062 participants in the 2011-2012 National Health and Nutrition Examination Survey (NHANES). The risk of TB infection in the deployed population was low-0.6% (95% confidence interval [CI]: 0.1-2.3%)-and was similar to the non-deployed population. The prevalence of latent TB infection (LTBI) in the U.S. population was not significantly different among deployed and non-deployed veterans and those with no military service. The limitations of these retrospective studies highlight the challenge in obtaining valid estimates of risk using retrospective data and the need for a more definitive study. Similar to civilian long-term travelers, risks for TB infection during deployment are focal in nature, and testing should be targeted to only those at increased risk.


[Identification of chemical constituents in Zhachong Shisanwei Pills by UPLC-Q-TOF-MS/MS].
Zhachong Shisanwei Pills, composed of 13 Chinese medicinal materials, are used for treating the diseases such as hemiplegia, pain of muscles and bones, rheumatism, and joint pain. The chemical composition and pharmacodynamics of Zhachong Shisanwei Pills have not been reported. Ultra-performance liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to quickly identify the chemical components of Zhachong Shisanwei Pills, which was performed with Shim-pack GIST C_(18) column(4.6 mm×150 mm, 5 μm). The gradient elution was conducted with methanol-0.05% acetic acid as the mobile phase. Electrospray ionization mass spectrometry(ESI-MS) was carried out in both positive and negative ion modes. The compounds were identidied based on accurate relative molecular weight, fragment ion species, and the MS data of reference substances and in literature. In conclusion, a total of 98 compounds were identified, including 19 organic acids, 36 flavonoids, 13 volatile oils, 8 tannins, 5 2-(2-phenylethyl)chromones, 5 amino acids, 3 sesquiterpenoids, 3 alkaloids, and 2 other compounds. This study characte-rized the chemical components of Zhachong Shisanwei Pills rapidly for the first time, laying a foundation for further research on the pharmacodynamic material basis and quality evaluation.


Development of a multi-pathogen oligonucleotide microarray for detection of Bacillus anthracis.
An oligonucleotide microarray system has been specifically designed to detect and differentiate Bacillus anthracis from other bacterial species present in clinical samples. The pilot-scale microarray initially incorporated probes to detect six common species of bacteria, which were fully evaluated. The microarray comprised long oligonucleotides (50--70-mer) designed to hybridise with the variable regions of the 16S rRNA genes. Probes which hybridised to virulence genes were also incorporated; for B. anthracis, these initially included the pag, lef, cap and vrrA (for partial genotyping) genes. Hybridisation conditions were initially optimised to be run using 5 x SSC, 0.1% SDS, 50 degrees C for 16 h. The detection limits of the microarray were determined under these conditions by titration of chromosomal DNA and unlabelled amplicons followed by hybridisation to determine the levels of sensitivity that could be obtained with the microarray. Two different amplification methodologies were also compared-specific-primer based PCR and random PCR (with the labelling stage incorporated). Higher sensitivity was obtained using specific PCR primers, however, since one of the desired outcomes of a microarray-based detection system was the high discrimination that it offered, random amplification and labelling was used as the amplification method of choice. The length of hybridisation was investigated using a time-course, and 1--2h was found to give optimal and higher signals than 16 h incubation. These results indicate that microarray technology can be employed in a diagnostic environment and moreover, results may be obtained in a similar time-scale to a standard PCR reaction, but with the advantage that no a priori knowledge of the infectious agent is required for detection.


Posttraumatic growth in non-Hispanic White and Hispanic women with cervical cancer.
This study examined posttraumatic growth (PTG) in Non-Hispanic White (NHW; n = 132) and Hispanic (HISP; n = 51) women who had been diagnosed with cervical cancer. Participants completed measures of PTG, spirituality, optimism, stage of cancer, and demographics variables. The results showed that women with cervical cancer reported PTG but at lower levels than in studies of women with breast cancer. Greater spirituality and more advanced cancer stage predicted more PTG, but optimism did not predict PTG. HISP women reported higher levels of PTG than NHW women, and greater spirituality in the HISP women partially accounted for the difference.


ChEBI in 2016: Improved services and an expanding collection of metabolites.
ChEBI is a database and ontology containing information about chemical entities of biological interest. It currently includes over 46,000 entries, each of which is classified within the ontology and assigned multiple annotations including (where relevant) a chemical structure, database cross-references, synonyms and literature citations. All content is freely available and can be accessed online at http://www.ebi.ac.uk/chebi. In this update paper, we describe recent improvements and additions to the ChEBI offering. We have substantially extended our collection of endogenous metabolites for several organisms including human, mouse, Escherichia coli and yeast. Our front-end has also been reworked and updated, improving the user experience, removing our dependency on Java applets in favour of embedded JavaScript components and moving from a monthly release update to a 'live' website. Programmatic access has been improved by the introduction of a library, libChEBI, in Java, Python and Matlab. Furthermore, we have added two new tools, namely an analysis tool, BiNChE, and a query tool for the ontology, OntoQuery.


Characterization of Fv-1 gene-product-mediated resistance transfer.
Fv-1 gene-specific resistance to ecotropic murine retroviruses can be transferred to permissive cells with RNA prepared from restrictive cells. The active RNA has a sedimentation coefficient of 20-22S at low ionic strength and multiple regions of activity (5-6S, 10-12S, 24-27S) at high ionic strength. The maximum level of resistance transfer cannot be enhanced with noninhibitory carrier RNA or facilitators of RNA uptake, and the maximum inhibition (40-60%) is independent of the MOI. The low efficiency of resistance transfer is, therefore, probably the result of a fraction of cells resistant to RNA uptake.


Differential effects of dibutyryl cyclic adenosine monophosphate and retinoic acid on the growth, differentiation, and cyclic adenosine monophosphate-binding protein of murine neuroblastoma cells.
Dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP) and beta-all-trans retinoic acid (RA) have been shown separately, and in some cases in combination, to modulate the growth, differentiation, and cAMP-dependent protein kinase (PK-A) activity of various tumor cells. The effects of Bt2cAMP and RA on a cholinergic clone (S20) of C1300 mouse neuroblastoma cells were explored in the present study. Treatment of these cells with 1 mM Bt2cAMP for 3 or more days resulted in 93% inhibition of cell proliferation in monolayer cultures and in 98% inhibition of colony formation in semisolid medium (0.5% agarose). In contrast, treatment of the cells with 1 or 10 microM RA had no inhibitory effects on cell proliferation in monolayer cultures but enhanced colony formation in agarose by up to 130%. The growth of cells treated with a combination of Bt2cAMP and RA was inhibited, although less so than with Bt2cAMP alone. Cells treated with Bt2cAMP alone or Bt2cAMP and RA extended long, neurite-like, cellular processes indicative of differentiation, whereas only a few untreated or RA-treated cells produced such extensions. The amount of [3H]cAMP-binding protein increased gradually up to 2-fold during a 3-day treatment with Bt2cAMP; in contrast it decreased by nearly 2-fold during RA treatment. These changes occurred in the level of the type I regulatory subunit (RI) of PK-A as determined by photoaffinity labeling with 8-azidoadenosine cyclic 3':5'-[32P]monophosphate. The increase in RI following Bt2cAMP treatment was corroborated by DEAE-cellulose chromatography. This analysis also demonstrated that type I PK-A is the predominant kinase in the untreated S20 cells and that RI exists as a free subunit in Bt2cAMP-treated cells. The activity of PK-A decreased by about 20% following treatment with either Bt2cAMP or RA and by 45% following treatment with a combination of both agents. These results suggest that the distinct effects of Bt2cAMP and RA on the anchorage-independent growth of S20 cells may be related to their opposite effects on the level of RI.


Investigating the effectiveness of Socially Assistive Robot on Depression and Cognitive Functions of Community Dwelling Older Adults with Cognitive Impairments.
We evaluated a socially assistive robot (SAR) named Hyodol during a six-week intervention. This study enrolled 69 older adults with cognitive decline. To screen the eligibility, we have used the following three criteria, namely Korean-Mini-Mental Status Exam score ≤ 26). Clinical Dementia Rating 0.5-2), and Diagnostics and Statistical Manual V. Participants were divided into three groups based on their cognitive function (i.e. very mild cognitive impairment (vMCI), mild cognitive impairment (MCI), and moderate cognitive impairment (MOCI). The groups were instructed to use Hyodol at home for a six-week period. Baseline and post-intervention surveys were performed after six weeks to examine the changes in perceived health, depression, and cognitive function. The vMCI group showed a reduction in the depression score after the intervention (t = -2.447, p = 0.040), in comparison to their peers in the control group. Further, the MCI group showed an improvement in the cognitive function score after the intervention (t = 2.690, p = 0.021). No significant improvement was found among MOCI participants who used the SARs. The significance of this study was to examine whether participants with different levels of cognitive functioning would diverge after a period of intervention using the Hyodol SARs. Moreover, it presented preliminary data for services and policies for home care treatment targeted to cognitive decline in older adults.


Resveratrol, a natural chemopreventive agent against degenerative diseases.
Resveratrol (3,5,4'-trihydroxystilbene) is a naturally occurring compound shown to modulate the risk of cardiovascular degenerative diseases (atherosclerosis) and inhibit chemical carcinogenesis in rodents. Various studies have demonstrated the effect of this phytoalexin on biological mechanisms involved in cardioprotection. These include modulation of lipid turnover, inhibition of eicosanoid production, prevention of the low-density lipoprotein oxidation and inhibition of platelet aggregation. Carcinogenesis in animal models can be divided at least into three stages: initiation, promotion and progression. Initiation occurs as result of interaction of a reactive form of carcinogen with DNA. Chemical carcinogens like polycyclic aromatic hydrocarbons are metabolized to reactive species by cytochrome P450 dependent enzymes activated through aryl hydrocarbon (Ah) receptor. The inhibition of tumor initiation by resveratrol most probably occurs through preventing the activation of Ah receptor. Resveratrol affects also several factors involved in tumor promotion and progression. Since tumor promoting agents alter the expression of genes whose products are associated with inflammation, chemoprevention of cardiovascular diseases and cancer may share the same common mechanisms. This includes principally modulation of the expression of growth factors and cytokines. Recently, chemopreventive properties of resveratrol have been associated with the inhibition of NF-kappaB. This transcription factor is strongly linked to inflammatory and immune responses, regulation of cell proliferation and apoptosis, thus it is important for tumor development and many other diseases including atherosclerosis. Although the mechanisms by which resveratrol interferes with the activation of NF-KB are not clear, it seems that inhibition of its degradation which is necessary for its cellular activation is the principal target. Based on the quantity and diversity of data available on the biological activity of resveratrol, it has to be considered a very promising chemoprotector and chemotherapeutic. Urgent investigations on its bioavailability and effects on in vivo systems, especially in humans, are necessary.


Essential concept of transplant immunology for clinical practice.
Our understanding of transplant immunology has advanced from gross allograft rejection to cellular response and to current molecular level. More sensitive assays have been developed to characterize patient sensitization and to detect pre-existing donor-specific antibodies (DSA) in pre-transplant crossmatch. After a transplant, pre-existing or de novo DSA are increasingly monitored to guide clinical management. Therefore, it is important for clinicians to understand the basic concepts and key components of transplant immunology as well as be familiarized with the modern immunological techniques used in kidney transplantation.


A Meta-Analysis of the Global Prevalence Rates of Staphylococcus aureus and Methicillin-Resistant S. aureus Contamination of Different Raw Meat Products.
Previous research has indicated that raw meats are frequently contaminated with Staphylococcus aureus, but data regarding the pooled prevalence rates of S. aureus and methicillin-resistant S. aureus (MRSA) contamination in different types of raw meat products (beef, chicken, and pork) and across different periods, regions, and purchase locations remain inconsistent. We systematically searched the PubMed, EMBASE, Ovid, Web of Science, and HighWire databases to identify studies published up to June 2016. The STROBE guidelines were used to assess the quality of the 39 studies included in this meta-analysis. We observed no significant differences in the pooled prevalence rates of S. aureus and MRSA contamination identified in various raw meat products, with overall pooled prevalence rates of 29.2% (95% confidence interval [CI], 22.8 to 35.9%) and 3.2% (95% CI, 1.8 to 4.9%) identified for the two contaminants, respectively. In the subgroup analyses, the prevalence of S. aureus contamination in chicken products was highest in Asian studies and significantly decreased over time worldwide. In European studies, the prevalence rates of S. aureus contamination in chicken and pork products were lower than those reported on other continents. The pooled prevalence rates of S. aureus contamination in chicken and pork products and MRSA contamination in beef and pork products were significantly higher in samples collected from retail sources than in samples collected from slaughterhouses and processing plants. These results highlight the need for good hygiene during transportation to and manipulation at retail outlets to reduce the risk of transmission of S. aureus and MRSA from meat products to humans.


DNAfan: a software tool for automated extraction and analysis of user-defined sequence regions.
DNAfan (DNA Feature ANalyzer) is a tool combining sequence-filtering and pattern searching. DNAfan automatically extracts user-defined sets of sequence fragments from large sequence sets. Fragments are defined by annotated gene feature keys and co- or non-occurring patterns within the feature or close to it. A gene feature parser and a pattern-based filter tool localizes and extracts the specific subset of sequences. The selected sequence data can subsequently be retrieved for analyses or further processed with DNAfan to find the occurrence of specific patterns or structural motifs. DNAfan is a powerful tool for pattern analysis. Its filter features restricts the pattern search to a well-defined set of sequences, allowing drastic reduction in false positive hits. http://bighost.ba.itb.cnr.it:8080/Framework.


Immunohistochemical markers of tumor prognosis in breast cancer in Egypt.
Although the relationship among different biologic markers of breast cancer has been shown to be important in predicting cancer behavior, expression of these markers can be an attribute of the population under study. Breast cancer is the most common malignancy among Egyptian women. We have studied a number of prognostic tumor markers in infiltrating ductal carcinoma in a group of Egyptian women and have correlated our results with traditional histologic parameters of behavior such as tumor nuclear grade and lymph node status. Seventy-five cases of infiltrating ductal breast cancer were evaluated from pathology archives. Formalin-fixed paraffin-embedded sections were immunohistochemically stained for PCNA, p53, c-erB-2, metallothionein, cathepsin-D, and GST-pi using specific antibodies and a standard avidin-biotin method. Most high-grade tumors were associated with higher PCNA expression and p53 abnormality. There was a significant difference between node-negative and node-positive tumors with regard to their metallothionein content; other markers, however, did not differ significantly between node-negative and node-positive tumors. PCNA expression, metallothionein expression, and p53 mutation appear to be markers of aggressive tumor behavior in Egyptian women with breast cancer.


Discovery of a novel hsp65 genotype within Mycobacterium massiliense associated with the rough colony morphology.
So far, genetic diversity among strains within Mycobacterium massiliense has rarely been studied. To investigate the genetic diversity among M. massiliense, we conducted phylogenetic analysis based on hsp65 (603-bp) and rpoB (711-bp) sequences from 65 M. massiliense Korean isolates. We found that hsp65 sequence analysis could clearly differentiate them into two distinct genotypes, Type I and Type II, which were isolated from 35 (53.8%) and 30 patients (46.2%), respectively. The rpoB sequence analysis revealed a total of four genotypes (R-I to R-IV) within M. massiliense strains, three of which (R-I, R-II and R-III) correlated with hsp65 Type I, and other (R-IV), which correlated with Type II. Interestingly, genotyping by the hsp65 method agreed well with colony morphology. Despite some exceptions, Type I and II correlated with smooth and rough colonies, respectively. Also, both types were completely different from one another in terms of MALDI-TOF mass spectrometry profiles of whole lipid. In addition, we developed PCR-restriction analysis (PRA) based on the Hinf I digestion of 644-bp hsp65 PCR amplicons, which enables the two genotypes within M. massiliense to be easily and reliably separated. In conclusion, two distinct hsp65 genotypes exist within M. massiliense strains, which differ from one another in terms of both morphology and lipid profile. Furthermore, our data indicates that Type II is a novel M. massiliense genotype being herein presented for the first time. The disparity in clinical traits between these two hsp65 genotypes needs to be exploited in the future study.


Botanicals and Phosphonate Show Potential to Replace Copper for Control of Potato Late Blight.
Potato late blight (PLB) caused by Phytophthora infestans (Pi) is the most harmful disease in potato production worldwide. In organic farming, copper is used despite its persistence in soil and toxicity to soil organisms. To replace copper, suspensions of powders from three promising botanicals, including bark of buckthorn (Frangula alnus, FA), roots of medicinal rhubarb (Rheum palmatum) and galls of the nutgall tree (Galla chinensis), were tested in multi-year field experiments. The current study shows for the first time that botanicals could replace copper under field conditions and best PLB reduction on leaves was achieved with FA, reaching a level close to that of 2 to 3 kg copper per hectare and year. Better results than with copper were achieved with Phosfik® (Ph), a phosphonate-based product. For both FA and Ph, the mode of action is based on induced resistance, for Ph also on direct fungicidal effects. A disadvantage of Ph is the accumulation of residues in potato tubers. Nevertheless, two to three applications with 2 to 3 L/ha of Ph would be feasible to not exceed a minimal risk level (MLR) of 20 mg/kg of phosphorous acid as proposed by the European Food Safety Authority. Due to an excellent environmental profile and a complex mode of action counteracting Pi resistance, phosphonate-based products would be most suitable for sustainable PLB management in integrated pest management (IPM) programmes.


Species specificity in the metabolism of nabilone. Relationship between toxicity and metabolic routes.
The disposition of 14C-nabilone has been examined in dogs and monkeys; it is rapidly absorbed and extensively metabolized in both species. A metabolic pathway initiated by the stereoselective enzymic reduction of the 9-keto moiety of nabilone was of major importance in the biotransformation of nabilone in the dog but was only a minor pathway in the monkey. The resulting long half-lived carbinol metabolites accounted for 77% of the circulating 14C in dog but for only 19% in monkey, 48 h after drug administration. Accumulation of carbinol metabolites in brain tissue was observed in dogs, with 24 h brain concentrations being five to six times higher than the plasma concentrations. No accumulation of carbinol metabolites, nabilone or of 14C occurred in the brain of monkeys. Accumulation of the carbinol metabolites in brain tissue has been implicated as the causative factor for the CNS toxicity observed in dogs treated chronically with nabilone. Comparison of nabilone pharmacokinetics in dog and humans showed the dog to be unique in producing high levels of carbinol metabolites, so that monkey appeared to be a more appropriate model than dog for pre-clinical toxicological and safety studies. Studies with liver 15,000 g supernatants indicated that the enzymic reduction of nabilone to its carbinol metabolite was a viable metabolic pathway in rat, dog and monkey, so that a distinct species difference exists between dog and monkey in the subsequent metabolism and/or elimination of these carbinol metabolites.


Non-Hermitian Chiral Heat Transport.
Exceptional point (EP) has been captivated as a concept of interpreting eigenvalue degeneracy and eigenstate exchange in non-Hermitian physics. The chirality in the vicinity of EP is intrinsically preserved and usually immune to external bias or perturbation, resulting in the robustness of asymmetric backscattering and directional emission in classical wave fields. Despite recent progress in non-Hermitian thermal diffusion, all state-of-the-art approaches fail to exhibit chiral states or directional robustness in heat transport. Here we report the first discovery of chiral heat transport, which is manifested only in the vicinity of EP but suppressed at the EP of a thermal system. The chiral heat transport demonstrates significant robustness against drastically varying advections and thermal perturbations imposed. Our results reveal the chirality in heat transport process and provide a novel strategy for manipulating mass, charge, and diffusive light.


Intraoperative Monitoring of Parathormone in the Surgical Treatment of Renal Hyperparathyroidism.
Intraoperative monitoring of parathyroid hormone can confirm the complete excision of hyperfunctional parathyroid tissue, as the plasma half-life of PTH is approximately 5 minutes. The purpose of this study was to analyse the values of parathormon (PTH) and the intraoperative impact in patients with secondary hyperparathyroidism of renal cause (sHPT). A series of 86 patients who were hospitalised in our clinic between February 2015 to December 2018, were included in the study rom. All patients underwent surgery with PTH monitoring. PTH was determined preoperatively, intraoperatively 15 minutes after parathyroidectomy and postoperatively. Out of a total of 86 patients, 6 patients had non-functional renal transplant. 81 patients were operated on per primam and 5 patients were operated for disease recurrence. There were 77 total parathyroidectomies and 4 subtotal parathyroidectomies. One patient had 5 parathyroid glands. There were 4 patients with recurrent hyper-plastic tissue excision. Blood samples were collected intraoperatively through the puncture of the jugular vein. The PTH value was determined by the Elecsys PTH STATÃÂî test. The mean value of preoperative PTH was 1658 pg / mL and decreased to 46.5 pg / mL at the end of the operation. Subsequently, the level of PTH harvested at 3-6 months increased slightly to 59.8 pg / mL. 80 (93%) of patients had elevated preoperative calcium values. Recurrent hyperparathyroidism was found in 1 of the 4 patients who underwent subtotal parathyroidectomy. IPTH value is influenced by the intraoperative manipulation of the parathyroid glands, the individual variability of PTH half-life and the physiological state of the patient. The decrease of PTH measured intraoperatively at 15 minutes after harvest with at least 90% of the preoperative value indicates the success of a total parathyroidectomy, with normalisation of calcium and PTH.


[Colorectal cancer--epidemiologic aspects].
Since 1971 the incidence of large bowel cancer in Austria has been higher than that of any other neoplastic disease. In 1983 it affected 3778 men and women, representing 14.8% of all malignancies. Concrete suggestions for an improvement of data collecting for the National Cancer Registry and the installation of a National Polyp Study are made. In the eastern parts of Austria the risk for the development of colorectal carcinoma is twice as high as in the western parts of the country. Environmental risk factors are discussed.


Value of clinical and echocardiographic features in predicting outcome in the fetus, infant, and child with tetralogy of Fallot with absent pulmonary valve complex.
We describe clinical and echocardiographic features of tetralogy of Fallot with absent pulmonary valve complex (TOF/APVC) and hypothesized that outcome might be related to pulmonary artery enlargement or severity of illness. We examined the clinical records of all 23 patients evaluated at our institution before death or surgical correction of TOF/APVC between 1990 and 2000. Echocardiograms for 16 patients (including 5 fetuses) were also reviewed, and measurements of the semilunar valves and pulmonary arteries were obtained and compared with patient's aortic annulus size and with established normal subjects. Actuarial survival was 15 of 23 patients (68%) at 4 years. Four fetuses were hydropic and none survived; 7 patients were ventilator dependent at operation and only 3 survived. No difference was noted in pulmonary artery diameters in survivors versus nonsurvivors. Pulmonary valve annulus size was larger in nonsurvivors (103 +/- 25% vs 71 +/- 24% of normal, p = 0.03); however, when fetal examinations were excluded, this difference did not persist. Thus, only hydrops and ventilator dependence at diagnosis predicted mortality. There was no correlation between postnatal measurements of pulmonary arteries and outcome. Larger pulmonary annulus size in hydropic fetuses and poor survival among patients diagnosed in utero suggests that the pathophysiology in TOF/APVC is not due entirely to the aneurysmal dilation of the pulmonary arteries but may be related to right-sided cardiac dysfunction.


Methotrexate iontophoresis versus coal tar ointment in palmoplantar psoriasis: A pilot study.
Palmoplantar psoriasis is often disabling and refractory to conventional therapy. Systemic drugs are indicated in its severe form, but side effects are a concern with their use. Methotrexate is one such systemic drug which is effective and cheap. To reduce systemic toxicity, methotrexate has been tried topically but results have been inconsistent due to poor drug penetration into the skin by passive diffusion. Iontophoresis may enhance its absorption and efficacy. To evaluate the efficacy and safety of topical methotrexate iontophoresis in comparison with coal tar ointment in the treatment of palmoplantar psoriasis. Thirty-one patients with palmar and/or plantar psoriasis were selected for the study and 28 patients completed it. The side having more severe involvement was treated while the other palm/sole served as a control. Iontophoresis using methotrexate solution was carried out on the study palm/sole with the injectable preparation of methotrexate (50 mg/2 ml) once a week for the first 4 weeks and subsequently every two weeks, for a total of six sittings. The control palm/sole was treated with coal tar ointment on other days. Erythema, scaling, induration and fissuring scores were noted in both groups before and after treatment. Both study and control groups showed decreases in scores but the reduction was more in the study group, the difference being statistically significant. Drawbacks of our study include the small sample size and the lack of follow-up. The study and control arms were not exactly matched and the study was not blinded. Methotrexate iontophoresis was safe and more effective than coal tar ointmentin palmoplantarpsoriasis.


Actinomadura rayongensis sp. nov., isolated from peat swamp forest soil.
A novel actinomycete strain RY35-68(T), isolated from a peat swamp forest soil sample in Rayong Province, Thailand, was characterized using a polyphasic approach. The strain belonged to the genus Actinomadura based on morphological and chemotaxonomic characteristics. Cell-wall analysis revealed the presence of meso-diaminopimelic acid and N-acetylmuramic acid in the peptidoglycan layer. The diagnostic sugar in whole-cell hydrolysates was identified as madurose. The predominant menaquinones were MK-9(H6), MK-9(H8) and MK-9(H4). The major cellular fatty acids were C16 : 0 and iso-C16 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. The genomic DNA G+C content was 73.7 mol%. On the basis of 16S rRNA gene sequence similarity analysis, strain RY35-68(T) was closely related to the species Actinomadura atramentaria JCM 6250(T) (97.5 %). The value of DNA-DNA relatedness between strain RY35-68(T) and A. atramentaria JCM 6250(T) was 37.6-42.6 %. On the basis of its phenotypic characteristics and these results mentioned, this strain could be distinguished from the closely related type strain and represents a novel species of the genus Actinomadura, for which the name Actinomadura rayongensis sp. nov. (type strain RY35-68(T) = JCM 19830(T) = TISTR 2211(T) = PCU 332(T)) is proposed.


The Mycobacterium avium complex - an underestimated threat to humans and animals.
The Mycobacterium avium complex (MAC) is a group of acid-resistant bacteria within the Mycobacteriaceae. Their cell walls have a specific structure impervious to many disinfectants. Mycobacteria are widespread in the environment and can also be found in food. This aim of the article is to review the current state of knowledge about the sources of infection, symptoms and treatment of MAC diseases in humans and animals, and summarizes the available methods for identifying the bacteria. It pays a special attention to the zoonotic potential of MAC bacteria and possible routes of transmission between humans and animals, including possible food-borne routes. MAC bacterial infections occur both in immunocompetent people and those with functional predispositions and compromised immunity, particularly during HIV infection or immunosuppressive treatment. The incidence of MAC infections in humans is growing, with the most common form of infection being pulmonary disease (MTC-PD); however, there are conflicting reports on the role of Mycobacterium avium paratuberculosis (MAP) in the development of Crohn's disease. MAC bacteria can also attack livestock, household pets, and wild animals. Unfortunately, treatment is lengthy and often fails due to microbiological relapse; there is also increasing evidence of MAC bacteria are developing multi-drug resistance. Although new antibiotics are being created to inhibit the growth and division of Mycobacterium avium, there is clearly a need for further research into the virulence factors associated with MAC bacteria. Further studies should also examine the role of MAP in the etiopathogenesis of Crohn's disease.


Rates of Reactions as a Mathematical Consequence of the Permanence of Atoms and the Role of Independent Reactions in the Description of Reaction Kinetics.
Linear algebra treatment of the permanence of atoms (mass conservation) naturally leads to the transformation of formation or destruction rates of components of a reaction mixture into rates of reaction steps, which are sufficient to describe the transformations mathematically. These steps form a scheme of independent reactions that can provide a rational basis for elucidating the reaction mechanism (network) while reducing both the component and parametric dimensionality of the description of kinetics. Several particular reaction examples are used to explain the method and show that rates of additional, dependent reactions cannot be unambiguously related to measured component rates. They also illustrate how the rates of dependent reactions can be correctly expressed in terms of the rates of independent reactions. The method starts only with a knowledge of the components of a reaction mixture. It is argued that the design of consistent reaction networks or mechanisms should take into account not only chemistry but also mathematics.


Interstitial Cajal-like cells (ICLC) as steroid hormone sensors in human myometrium: immunocytochemical approach.
Expression of estrogen (ER) and progesterone (PR) receptors was investigated in cultured human normal myometrial cells (non-pregnant uterus, fertile period). The ER and PR expression was studied by immunohistochemistry and immunofluorescence on either myocytes or interstitial Cajal-like cells (ICLC). Only those cells double immunostained for c-kit and steroid receptors were considered as ICLC. ER and/or PR immunoreactivity was localized in ICLC, primarily concentrated at the nucleus level, but it was also observed in the cell body (cytoplasm) and processes. Stronger immunopositive reaction in the ICLC nucleus for PR than for ER was noted. Under our experimental conditions, a clear positive repeatable reaction for steroid receptors could not be detected in myocytes. In conclusion, these data suggest that ICLC could be true hormonal 'sensors', possibly participating in the regulation of human myometrial contractions (via gap junctions with myocytes and/or by paracrine signaling).


Health-related quality of life at median 12 months after aneurysmal subarachnoid hemorrhage, measured with EuroQoL-5D.
A measurement of quality of life (QoL) should cover the important aspects of daily life and be easy to perform. Ease of performance is especially important for patients with spontaneous subarachnoid haemorrhage (SAH), since fatigue and cognitive disabilities are known sequeles. EuroQoL (EQ-5D) is a preference-based instrument measuring QoL, based on self-reported health status in five dimensions: Mobility, Self-Care, Usual Activities, Pain/Discomfort and Anxiety/Depression. In the present study EuroQoL was used in patients with aneurysmal SAH (aSAH) in comparison with a Swedish reference population. We also determined the extent to which demographic characteristics and clinical parameters predicted outcome. Seven hundred fifty-five patients with aSAH were studied after a median 12 months. The proportion of patients in the best QoL category for each dimension was compared with the corresponding proportion in an age matched reference population. Disease severity was measured using the World Federation of Neurosurgical Societies' SAH grading system and the Fisher scale. The extent to which demographic and clinical factors predicted outcome was evaluated using linear regression. Aneurysmal SAH patients generally had a worse QoL compared with the reference population, in all five dimensions of EQ-5D. In the patient population, disease severity predicted worse outcome in all five dimensions. Female gender and surgery as treatment method (in the case of anterior aneurysms) predicted worse outcome in Usual Activities and Anxiety/Depression. The nature of the sequeles after SAH depends on severity of disease, gender and treatment method. These factors should be more emphasised in planning rehabilitation.


Urogenital disorders associated with oestrogen deficiency: the role of promestriene as topical oestrogen therapy.
Urogenital disorders associated with oestrogen deficiency affect many women throughout menopausal transition. Symptoms such as vaginal dryness, burning, pruritus, dyspareunia, urinary tract urgency/frequency and incontinence have a significant impact on the individual's quality of life. For younger and healthy menopausal women, systemic oestrogen replacement may improve both vasomotor and urogenital symptoms and will be the treatment of choice. However, a proportion of women on systemic therapy still experience symptoms associated with urogenital atrophy, and patients with oestrogen-dependent cancers may be at risk from systemic oestrogen replacement. For women with mainly urogenital symptoms, local oestrogen is a logical choice and it is often more effective than systemic hormone replacement therapy. Generally speaking, there are no contraindications to local therapy. In terms of which topical preparation to use, a wide range of products are available. Promestriene is an analogue of oestradiol which is minimally absorbed and it has been shown to be effective in reversing atrophic changes caused by oestrogen deficiency in women undergoing natural or surgically induced menopause. Given the absence of systemic activity, promestriene may be a good choice in women requiring purely locally oestrogen, and those who have survived, or who are at risk of breast cancer and who have severe vulvo-vaginal symptoms.


Characterization of anaerobic heterotrophic bacteria isolated from freshwater lake sediments.
Strict anaerobic culture techniques were used to quantitatively and qualitatively evaluate the anaerobic heterotrophic bacteria present at the sediment-water interface of hyperutrophic Wintergreen Lake (Augusta, Mich.). Anaerobic plate counts remained constant from March through December, 1973, ranging from 2.4 X 10(6) to 5.7 X 10(6) organisms/g (dry weight) of sediment. The isolatable bacteria represented a small percentage of the total microbial community, which was shown by direct microscopic counts to be 2.0 X 10'' organisms/g (dry weight) of sediment during June and July. Bacteria of the genus Clostridium dominated the isolates obtained, accounting for 71.8% of the 960 isolates examined. A single species, Clostridium bifermentens, comprised 47.7% of the total. Additional bacterial groups and the percentage in which they were isolated included: Streptococcus sp. (10.8%), unidentified curved rods (9.5%y, gram-positive nonsporing rods (5.6%), and motile gram-negative rods (1.9%). Temperature growth studies demonstrated the ability of all the isolates to grow at in situ sediment temperatures. Gas-liqid radiochromatography was used to determine the soluble metabolic end products from [U-14C]glucose and a U-14C-labeled amino acid mixture by representative sedimentary clostridial isolates and by natural sediment microbial communities. At in situ temperatures the natural sediment microflora produced soluble fermentative end products characteristic of those elaborated by the clostridial isolates tested. These results are considered strong presumptive evidence that clostridia are actively metabolizing in the sediments of Wintergreen Lake.


The multi-drug efflux system AcrABZ-TolC is essential for infection of Salmonella Typhimurium by the flagellum-dependent bacteriophage Chi.
Bacteriophages are the most abundant biological entities in the biosphere. Due to their host specificity and ability to kill bacteria rapidly, bacteriophages have many potential healthcare applications, including therapy against antibiotic-resistant bacteria. Infection by flagellotropic bacteriophages requires a properly rotating bacterial flagellar filament. The flagella-dependent phage χ (Chi) infects serovars of the pathogenic enterobacterium Salmonella enterica However, cell surface receptors and proteins involved in other stages of χ infection have not been discovered to date. We screened a multi-gene deletion library of S. enterica serovar Typhimurium by spotting mutants on soft agar plates seeded with bacteriophage χ and monitoring their ability to grow and form a swim ring, a characteristic of bacteriophage-resistant motile mutants. Those multi-gene deletion regions identified to be important for χ infectivity were further investigated by characterizing the phenotypes of corresponding single-gene deletion mutants. This way, we identified motile mutants with varying degrees of resistance to χ. Deletions in individual genes encoding the AcrABZ-TolC multi-drug efflux system drastically reduced infection by bacteriophage χ. Furthermore, an acrABtolC triple deletion strain was fully resistant to χ. Infection was severely reduced but not entirely blocked by the deletion of the gene tig encoding the molecular chaperone trigger factor. Finally, deletion in genes encoding enzymes involved in the synthesis of the antioxidants glutathione (GSH) and uric acid resulted in reduced infectivity. Our findings begin to elucidate poorly understood processes involved in later stages of flagellotropic bacteriophage infection and informs research aimed at the use of bacteriophages to combat antibiotic-resistant bacterial infections.IMPORTANCEAntimicrobial resistance is a large concern in the healthcare field. With more multi-drug resistant bacterial pathogens emerging, other techniques for eliminating bacterial infections are being explored. Among these is phage therapy, where combinations of specific phages are used to treat infections. Generally, phages utilize cell appendages and surface receptors for the initial attachment to their host. Phages that are flagellotropic are of particular interest because flagella are often important in bacterial virulence, making resistance to attachment of these phages harder to achieve without reducing virulence. This study discovered the importance of a multi-drug efflux pump for the infection of Salmonella enterica by a flagellotropic phage. In theory, if a bacterial pathogen develops phage resistance by altering expression of the efflux pump then the pathogen would simultaneously become more susceptible to the antibiotic substrates of the pump. Thus, co-administering antibiotics and flagellotropic phage may be a particularly potent antibacterial therapy.


Assessment of Prevalence of Hypertension and its Risk Factors among Adult Tribal Population Residing in Lohandiguda Block of Bastar District of Chhattisgarh.
Hypertension is known as the silent killer. It comprehends the top rank in non-infectious disease amongst adults; accountable for the deaths every year across the world. It is essential to consider the individual impact of risk factors and their impact on hypertension. This study thus elicited the socio-demographic characteristics, the prevalence of hypertension and associated risk factors, and its impact on adults with hypertension. To estimate the hypertension prevalence and its associated risk factors among adult tribal populations aged 25-60 years residing in Lohandiguda block of Bastar district of Chhattisgarh. A community-based cross-sectional analytical study was used and the setting was done at the field practice area under the three primary health centers of Lohandiguda block, Bastar district of Chhattisgarh. It was carried out among 330 adult tribes residing for ≥1 year in the present locality. Data was collected by door-to-door visits through pre-designed, pretested, semi-structured questionnaire via face-to-face interview method and anthropometric measurement was done by using standard guidelines. The sampling method was multistage sampling. IBM SPSS STATISTICS-20.0 (IBM Corp., Armonk, NY, USA) software. The overall prevalence of pre-hypertension and hypertension among tribal subjects was 34.9% and 47.3%, respectively. Of total hypertensive 27.3% were having stage-1 hypertension, 13.9% were having stage-2 hypertension and 6.0% were already diagnosed cases. Risk factors found in multivariate analysis are occupation (unemployed 0.012), frequency of smokeless tobacco used per day (0.,017) and central obesity (0.000). As hypertension is a multi-factorial disease the study found strong predictors like occupation, frequency of smokeless tobacco per day and having central obesity with significant difference.


Extracellular Collagen Promotes Interleukin-1β-Induced Urokinase-Type Plasminogen Activator Production by Human Corneal Fibroblasts.
Keratocytes maintain homeostasis of the corneal stroma through synthesis, secretion, and degradation of collagen fibrils of the extracellular matrix. Given that these cells are essentially embedded in a collagen matrix, keratocyte-collagen interactions may play a key role in regulation of the expression or activation of enzymes responsible for matrix degradation including urokinase-type plasminogen activator (uPA), plasmin, and matrix metalloproteinases (MMPs). We examined the effect of extracellular collagen on the production of uPA by corneal fibroblasts (activated keratocytes) stimulated with the proinflammatory cytokine interleukin-1β (IL-1β). Human corneal fibroblasts were cultured either on plastic or in a three-dimensional gel of type I collagen. Plasminogen activators were detected by fibrin zymography, whereas the IL-1 receptor (IL-1R) and MMPs were detected by immunoblot analysis. Collagen degradation by corneal fibroblasts was assessed by measurement of hydroxyproline in acid hydrolysates of culture supernatants. Collagen and IL-1β synergistically increased the synthesis and secretion of uPA in corneal fibroblasts. Collagen also upregulated IL-1R expression in the cells in a concentration-dependent manner. The conversion of extracellular plasminogen to plasmin, as well as the plasminogen-dependent activation of MMP-1 and MMP-3 and degradation of collagen apparent in three-dimensional cultures of corneal fibroblasts exposed to IL-1β, were all abolished by a selective uPA inhibitor. Collagen and IL-1β cooperate to upregulate uPA production by corneal fibroblasts. Furthermore, IL-1β-induced collagen degradation by these cells appears to be strictly dependent on uPA expression and mediated by a uPA-plasmin-MMP pathway.


The Effects of Modified Constraint-Induced Movement Therapy in Acute Subcortical Cerebral Infarction.
Background: Constraint-induced movement therapy (CIMT) promotes upper extremity recovery post stroke, however, it is difficult to implement clinically due to its high resource demand and safety of the restraint. Therefore, we propose that modified CIMT (mCIMT) be used to treat individuals with acute subcortical infarction. Objective: To evaluate the therapeutic effects of mCIMT in patients with acute subcortical infarction, and investigate the possible mechanisms underlying the effect. Methods: The role of mCIMT was investigated in 26 individuals experiencing subcortical infarction in the preceding 14 days. Patients were randomly assigned to either mCIMT or standard therapy. mCIMT group was treated daily for 3 h over 10 consecutive working days, using a mitt on the unaffected arm for up to 30% of waking hours. The control group was treated with an equal dose of occupational therapy and physical therapy. During the 3-month follow-up, the motor functions of the affected limb were assessed by the Wolf Motor Function Test (WMFT) and Motor Activity Log (MAL). Altered cortical excitability was assessed via transcranial magnetic stimulation (TMS). Results: Treatment significantly improved the movement in the mCIMT group compared with the control group. The mean WMF score was significantly higher in the mCIMT group compared with the control group. Further, the appearance of motor-evoked potentials (MEPs) were significantly higher in the mCIMT group compared with the baseline data. A significant change in ipsilesional silent period (SP) occurred in the mCIMT group compared with the control group. However, we found no difference between two groups in motor function or electrophysiological parameters after 3 months of follow-up. Conclusions: mCIMT resulted in significant functional changes in timed movement immediately following treatment in patients with acute subcortical infarction. Further, early mCIMT improved ipsilesional cortical excitability. However, no long-term effects were seen.


Multisite clinical evaluation of a rapid test for Entamoeba histolytica in stool.
Rapid point-of-care detection of enteric protozoa in diarrheal stool is desirable in clinical and research settings to efficiently determine the etiology of diarrhea. We analyzed the ability of the third-generation E. histolytica Quik Chek assay developed by Techlab to detect amebic antigens in fecal samples collected from independent study populations in South Africa and Bangladesh. We compared the performance of this recently released rapid test to that of the commercially available ProSpecT Entamoeba histolytica microplate assay from Remel and the E. histolytica II enzyme-linked immunosorbent assay (ELISA) from Techlab, using real-time and nested-PCR for Entamoeba species to resolve any discrepant results. After discrepant resolution, The E. histolytica Quik Chek assay exhibited sensitivity and specificity compared to the E. histolytica II ELISA of 98.0% (95% confidence interval [CI], 92.9% to 99.8%) and 100% (95% CI, 99.0% to 100%), respectively. Compared to the ProSpecT microplate assay, the E. histolytica Quik Chek (Quik Chek) assay exhibited 97.0% sensitivity (95% CI, 91.5% to 99.4%) and 100% specificity (95% CI, 99.0% to 100%). Our results indicate that the Quik Chek is a robust assay for the specific detection of E. histolytica trophozoites in unfixed frozen clinical stool samples.


Histopathological findings in appendectomy specimens: a retrospective clinicopathological analysis.
The prevalence and clinical features of the various obstructive lesions in appendectomy specimens studied pathologically with special emphasis on unusualetiological findings were evaluated. The clinic-pathological data of 251 patients who underwent appendectomies for presumed acute appendicitis from January; 2008 to December; 2010; were reviewed retrospectively. Among all appendectomies performed, eleven (4.38%) specimens revealed incidental abnormal pathological diagnoses: Four cases (1.59%) of Enterobiusvermicularis, Ascaris lumbricoides two cases (0.79 %), schistosomiasis eggs in three cases (1.19%), and parasitic amebiasis in two cases (0.79%). None suffered tumor in any appendectomies specimens. Mean age was lower, perforated and gangrenous appendicitis was absent and negative appendectomy rate was higher in patients with unusual appendix pathology.


Method to generate highly stable D-amino acid analogs of bioactive helical peptides using a mirror image of the entire PDB.
Biologics are a rapidly growing class of therapeutics with many advantages over traditional small molecule drugs. A major obstacle to their development is that proteins and peptides are easily destroyed by proteases and, thus, typically have prohibitively short half-lives in human gut, plasma, and cells. One of the most effective ways to prevent degradation is to engineer analogs from dextrorotary (D)-amino acids, with up to 105-fold improvements in potency reported. We here propose a general peptide-engineering platform that overcomes limitations of previous methods. By creating a mirror image of every structure in the Protein Data Bank (PDB), we generate a database of ∼2.8 million D-peptides. To obtain a D-analog of a given peptide, we search the (D)-PDB for similar configurations of its critical-"hotspot"-residues. As a proof of concept, we apply our method to two peptides that are Food and Drug Administration approved as therapeutics for diabetes and osteoporosis, respectively. We obtain D-analogs that activate the GLP1 and PTH1 receptors with the same efficacy as their natural counterparts and show greatly increased half-life.


Flu, Floods, and Fire: Ethical Public Health Preparedness.
Even as public health ethics was developing as a field, major incidents such as 9/11 and the SARS epidemic propelled discourse around public health emergency preparedness and response. Policy and practice shifted to a multidisciplinary approach, recognizing the broad range of potential threats to public health, including biological, physical, radiological, and chemical threats. This propelled the development of surveillance systems to detect incidents, laboratory capacities to rapidly test for potential threats, and therapeutic and social countermeasures to prepare for and respond to a range of hazards. In bringing public health ethics and emergency preparedness together, Emergency Ethics: Public Health Preparedness and Response adds depth and complexity to both fields. As global threats continue to emerge, the book, edited by Bruce Jennings, John D. Arras, Drue H. Barrett, and Barbara A. Ellis, will offer a vital compass.


Increased plasma and pituitary prolactin concentrations in adult male rats with selective elevation of FSH levels may be explained by reduced testosterone and increased estradiol production.
The roles of testosterone and estradiol in regulating prolactin concentrations were studied in acutely castrated adult male rats receiving subcutaneous Silastic implants of the sex steroids. Testosterone was administered in increasing doses, from subphysiologic to intact levels, both alone and in combination with a small, single dose of estradiol. The study was designed to assess whether a change in the relative rates of sex steroid production could account for an increase in PRL release in the absence of other testicular factors. At very low levels of plasma testosterone, FSH and LH levels were indistinguishable from castrate controls. As plasma testosterone concentration increased, both plasma FSH and LH levels were suppressed progressively to intact levels. When a subphysiologic dose of testosterone was coadministered with a small dose of estradiol, the combined effects produced a midcastrate level of FSH but maintained a normal level of LH similar to the selective increase in FSH concentration observed in men with germinal aplasia. Although PRL levels were indistinguishable in intact and castrate controls, testosterone replacement by capsule increased prolactin in a dose-related manner so that, at the physiologic level of testosterone, prolactin was elevated two-fold (P less than 0.01), similar to the level achieved with estradiol replacement alone. Pituitary prolactin levels also increased with increasing doses of testosterone but values remained within the range measured in intact controls. When estradiol was coadministered with testosterone, the combination produced different effects depending on the testosterone dose.(ABSTRACT TRUNCATED AT 250 WORDS)


Targeted protection of hepatocytes from galactosamine toxicity in vivo.
We present an in vivo model for specific protection of normal hepatocytes from damage by the highly specific hepatotoxin galactosamine. The idea is based on the fact that normal, unlike malignant, hepatocytes possess unique cell-surface receptors that can bind and internalize galactose terminal (asialo)glycoproteins by receptor-mediated endocytosis. A targetable carrier-antagonist conjugate was formed by coupling asialofetuin to the galactosamine antagonist uridine monophosphate. Intravenous injection of the antagonist conjugate resulted in specific uptake by the liver. Rats treated with carrier-antagonist conjugate together with a toxic dose of galactosamine developed significantly less hepatotoxicity than did controls. We conclude that a galactosamine antagonist can be targeted to liver, resulting in specific protection of hepatocytes from galactosamine toxicity in vivo. Because hepatoma cells lack asialoglycoprotein receptor activity, this "targeted rescue" may be of value in the differential protection of normal cells in the treatment of hepatocellular carcinoma.


Genetic and clinical characteristics of multiple endocrine neoplasia type 1.
Multiple endocrine neoplasia type 1 (MEN 1) is an inherited disorder of autosomal-dominant type encompassing tumors of the parathyroid glands, anterior pituitary and endocrine pancreas. The genetic defect responsible for MEN 1 maps to the long arm of chromosome 11 (11q13). Constitutional heterozygosity for the MEN 1 region is lost in proliferating pancreatic endocrine and parathyroid tissue, which suggests that the MEN 1 gene belongs to the group of tumor suppressor genes (antioncogenes). Genetic screening by linkage analysis using RFLP markers allows selection of gene carriers with high degree of accuracy and makes it possible to concentrate the laborious biochemical screening efforts to family members at risk of developing the MEN 1 syndrome. Biochemical screening of the MEN 1 lesions in young asymptomatic individuals decreases the age at diagnosis with many years (decades) and therapeutic intervention can be instituted early. Although it still remains to be established whether early intervention will have an impact on mortality, our long-term experience with 20 MEN 1 kindreds suggests a reduced morbidity as a result of thorough biochemical screening.


Inhibitory effect of single and repeated doses of nilotinib on the pharmacokinetics of CYP3A substrate midazolam.
Effects of single and repeated doses of nilotinib on the pharmacokinetics of midazolam, a cytochrome P450 3A (CYP3A) substrate, were assessed in 2 separate studies. In the single-dose nilotinib study, 18 healthy subjects were randomized to 6 treatment sequences to receive single dose of nilotinib 600 mg, midazolam 4 mg, and coadministration of both in a crossover manner. In the repeated-dose nilotinib study, 19 chronic myeloid leukemia patients took a single dose of midazolam 2 mg on days 1 and 13, and nilotinib 400 mg twice daily from days 2-13. In the single-dose study, the geometric mean ratio of the area under the plasma concentration time curve extrapolated to infinity (AUC(inf)) of midazolam plus nilotinib vs. midazolam was 1.3 (90%CI, 1.2-1.5) and the maximum observed serum concentration (C(max)) was 1.2 (90%CI, 1.0-1.4). In the repeated-dose study, the values for AUC(inf) and C(max) were 2.6 (90%CI, 2.1-3.3) and 2.0 (90%CI, 1.7-2.4), respectively. These results indicate that single-dose and repeated-dose administration of nilotinib results in weak and moderate inhibition of CYP3A, respectively. Therefore, appropriate monitoring and dose adjustment may be needed for drugs that are mainly metabolized by CYP3A, and have narrow therapeutic index, when coadministered with nilotinib.


Environmental and occupational pesticide exposure and human sperm parameters: a systematic review.
Of continuing concern are the associations between environmental or occupational exposures to pesticides and semen quality parameters. Prior research has indicated that there may be associations between exposure to pesticides of a variety of classes and decreased sperm health. The intent of this review was to summarize the most recent evidence related to pesticide exposures and commonly used semen quality parameters, including concentration, motility and morphology. The recent literature was searched for studies published between January 2007 and August 2012 that focused on environmental or occupational pesticide exposures. Included in the review are 17 studies, 15 of which reported significant associations between exposure to pesticides and semen quality indicators. Two studies also investigated the roles genetic polymorphisms may play in the strength or directions of these associations. Specific pesticides targeted for study included dichlorodiphenyltrichloroethane (DDT), hexachlorocyclohexane (HCH), and abamectin. Pyrethroids and organophosphates were analyzed as classes of pesticides rather than as individual compounds, primarily due to the limitations of exposure assessment techniques. Overall, a majority of the studies reported significant associations between pesticide exposure and sperm parameters. A decrease in sperm concentration was the most commonly reported finding among all of the pesticide classes investigated. Decreased motility was also associated with exposures to each of the pesticide classes, although these findings were less frequent across studies. An association between pesticide exposure and sperm morphology was less clear, with only two studies reporting an association. The evidence presented in this review continues to support the hypothesis that exposures to pesticides at environmentally or occupationally relevant levels may be associated with decreased sperm health. Future work in this area should focus on associations between specific pesticides or metabolic products and sperm quality parameters. Analysis of effects of varying genetic characteristics, especially in genes related to pesticide metabolism, also needs further attention.


Mechanisms of hexameric helicases.
Ring-shaped hexameric helicases are essential motor proteins that separate duplex nucleic acid strands for DNA replication, recombination, and transcriptional regulation. Two evolutionarily distinct lineages of these enzymes, predicated on RecA and AAA+ ATPase folds, have been identified and characterized to date. Hexameric helicases couple NTP hydrolysis with conformational changes that move nucleic acid substrates through a central pore in the enzyme. How hexameric helicases productively engage client DNA or RNA segments and use successive rounds of NTPase activity to power translocation and unwinding have been longstanding questions in the field. Recent structural and biophysical findings are beginning to reveal commonalities in NTP hydrolysis and substrate translocation by diverse hexameric helicase families. Here, we review these molecular mechanisms and highlight aspects of their function that are yet to be understood.


Metal-free methylation of a pyridine N-oxide C-H bond by using peroxides.
Metal-free methylation of a pyridine N-oxide C-H bond was developed using peroxide as a methyl reagent under neat conditions. Pyridine N-oxide derivatives with various groups (e.g., Cl, NO2, and OCH3) were all suitable substrates, and the desired products were obtained in moderate to excellent yields under standard conditions. Moreover, the methylation can be performed with a good yield on the gram-scale experiment. Tentative mechanistic studies show that the methylation is a classical radical process.


Fetuin-null mice are protected against obesity and insulin resistance associated with aging.
alpha2-HS glycoprotein (AHSG), also known as fetuin-A, inhibits insulin receptor autophosphorylation and tyrosine kinase activity in vitro and in vivo. Earlier we have shown that fetuin-null (KO) mice demonstrate improved insulin sensitivity and resistance to diet-induced obesity. Since aging is associated with insulin resistance and impaired glucose handling, we tested the hypothesis that fetuin-null (KO) mice are resilient to changes in insulin sensitivity associated with aging. Aged (80-week-old) fetuin-null mice were leaner and demonstrated significantly lower body weights compared to age- and sex-matched wild-type (WT) littermates. Leanness in aged fetuin KO mice was accompanied by a significant increase in dark-onset energy expenditure, without marked alteration of respiratory quotient. In comparison to WT mice, fetuin KO mice demonstrated a lower fasting insulin resistance index, and significantly lower blood glucose and insulin levels, following a 4h fast. Interestingly, despite significantly decreased insulin levels during a glucose tolerance test, aged fetuin-null mice demonstrated a similar glucose excursion as WT mice, indicative of improved insulin sensitivity. Analysis of aldehyde-fuchsin stained pancreas from aged fetuin KO mice indicated no difference in islet beta-cell size or number. An insulin tolerance test confirmed the increased insulin sensitivity of aged fetuin KO mice. Further, compared to WT mice, aged fetuin-null mice demonstrated increased skeletal muscle and liver IR autophosphorylation and TK activity. Taken together, this study suggests that the absence of fetuin may contribute to the improvement of insulin sensitivity associated with aging.


Substrate specificity of cellobiose dehydrogenase from Phanerochaete chrysosporium.
Substrate structural mapping suggests that the catalytic site of cellobiose dehydrogenase from Phanerochaete chrysosporium forms a narrow cave with two hexose binding subsites. Kinetic data also show that beta-di or oligosaccharides are favored electron donors with respect to both KM and kcat. Surprisingly, thiocellobiose showed an even higher kcat than cellobiose, although the KM value was somewhat higher. The CDH was purified using an updated protocol.


Long-Term Efficacy of Methylphenidate Immediate-Release for the Treatment of Childhood ADHD.
To evaluate the long-term effects of methylphenidate imediate-release (MPH-IR), and to confirm the efficacy established in previous meta-analyses of short-term studies. Published and unpublished studies in which participants were treated with MPH-IR for 12 weeks or more were searched. Pooled effect sizes from these studies were computed with the DerSimonian and Laird random-effect model. Meta-regression analysis was conducted to estimate covariates associated with treatment effects. Seven studies were included. Pooled parents ratings for inattention and hyperactivity/impulsivity resulted in standardized mean difference (SMD) = 0.96 (95% confidence interval [CI] = [0.60, 1.32]) and SMD = 1.12 (95% CI = [0.85, 1.39]), respectively; pooled teachers ratings showed SMD = 0.98 (95% CI = [0.09, 1.86]) for inattention and SMD = 1.25 (95% CI = [0.7, 1.81]) for hyperactivity/impulsivity. No evidence of association of any covariates with treatment effect was detected in the meta-regression. MPH-IR is efficacious for childhood ADHD for periods longer than 12 weeks.