Sociocultural and behavioral influences on weight gain during pregnancy.
Studies have consistently identified a positive association between prenatal weight gain and birth weight. Much less, however, is known about factors that may influence women to gain weight within currently recommended ranges. The importance of this issue is suggested by recent reports indicating that only 30-40% of women actually gain weight within these ranges. This paper examines demographic, sociocultural, and behavioral factors that are associated with, and may influence risk of, low prenatal weight gain among adult women with low and normal body mass indexes. Available data suggest that these factors include ethnicity, socioeconomic status, age, education, pregnancy intendedness or wantedness, prenatal advice, and psychosocial characteristics such as attitude toward weight gain, social support, depression, stress, anxiety, and self-efficacy. Potential theoretical models for these associations include biological, behavioral, and mixed pathways. The design of targeted intervention studies will depend on further identification and characterization of sociocultural and behavioral risk factors that, along with reproductive and nutritional characteristics, may predict which women are most likely to have inadequate prenatal weight gain.


Empirical comparisons of multiple Mendelian randomization approaches in the presence of assortative mating.
Mendelian randomization (MR) is widely used to unravel causal relationships in epidemiological studies. Whereas multiple MR methods have been developed to control for bias due to horizontal pleiotropy, their performance in the presence of other sources of bias, like non-random mating, has been mostly evaluated using simulated data. Empirical comparisons of MR estimators in such scenarios have yet to be conducted. Pleiotropy and non-random mating have been shown to account equally for the genetic correlation between height and educational attainment. Previous studies probing the causal nature of this association have produced conflicting results. We estimated the causal effect of height on educational attainment in various MR models, including the MR-Egger and the MR-Direction of Causation (MR-DoC) models that correct for, or explicitly model, horizontal pleiotropy. We reproduced the weak but positive association between height and education in the Netherlands Twin Register sample (P= 3.9 × 10-6). All MR analyses suggested that height has a robust, albeit small, causal effect on education. We showed via simulations that potential assortment for height and education had no effect on the causal parameter in the MR-DoC model. With the pleiotropic effect freely estimated, MR-DoC yielded a null finding. Non-random mating may have a bearing on the results of MR studies based on unrelated individuals. Family data enable tests of causal relationships to be conducted more rigorously, and are recommended to triangulate results of MR studies assessing pairs of traits leading to non-random mate selection.


Chronic statin therapy is associated with enhanced cutaneous vascular responsiveness to sympathetic outflow during passive heat stress.
Impairments in both central sympathetic and peripheral microvascular function contribute to blunted reflex cutaneous vasodilatation during heat stress in healthy older adults. Hypercholesterolaemia is associated with decrements in neurovascular function; however, little is known about the impact of hypercholesterolaemia on the integrated responses to heat stress. Further, whether chronic statin therapy alters skin sympathetic outflow or its relation to cutaneous vascular conductance during heat stress is unknown. We demonstrate that reflex cutaneous vasodilatation is impaired in older hypercholesterolaemic adults but not in formerly hypercholesterolaemic adults currently treated with a statin compared to age-matched controls. Additionally, chronic statin treatment-induced improvements in reflex vasodilatation are mediated, in part, by increases in end-organ responsiveness to efferent sympathetic outflow during whole-body heating. These data add to the growing body of literature substantiating the beneficial pleiotropic neurovascular effects of chronic statin treatment and provide further support for the use of statins to confer additional cardioprotective benefits in older adults. Attenuated reflex cutaneous vasodilatation in healthy human ageing is mediated by alterations in both central (sympathetic outflow) and peripheral (microvascular endothelial) function. Hypercholesterolaemia is associated with further impairments in neurovascular function. HMG-CoA reductase inhibitors (statins) improve cutaneous endothelium-dependent dilatation; however, whether statin therapy alters skin sympathetic nervous system activity (SSNA) or its relation to cutaneous vascular conductance (CVC) during passive heat stress is unknown. We hypothesized that (1) hypercholesterolaemic older adults would demonstrate blunted increases in both SSNA and CVC during passive heating and (2) chronic statin treatment would improve the response range and sensitivity of the SSNA:CVC relation. Reflex vasodilatation in response to a 1.0°C rise in oral temperature (Tor ; water perfused suit) was induced in 13 healthy normocholesterolaemic adults (62 ± 2 years; LDL = 113 ± 7 mg/dl), 10 hypercholesterolaemic adults (60 ± 1 years; LDL = 183 ± 2 mg/dl), and 10 previously hypercholesterolaemic adults (64 ± 1 years; LDL = 102 ± 2 mg/dl) treated with lipophilic statin (10-40 mg daily). SSNA (peroneal microneurography) and red cell flux (laser-Doppler flowmetry) in the innervated dermatome (dorsum of foot) were continuously measured. Reflex vasodilatation was blunted in hypercholesterolaemic adults, but not in statin-treated adults, compared to normocholesterolaemic adults (at ∆Tor = 1.0°C: normal = 36 ± 1%CVCmax , high = 32 ± 1%CVCmax , statin = 38 ± 1%CVCmax ; P < 0.01). ∆SSNA was not different (at ∆Tor = 1.0°C: normal: ∆ = 393 ± 96%, high: ∆ = 311 ± 120%, statin: ∆ = 256 ± 90%; P = 0.11). The slope of the SSNA:CVC relation was blunted in hypercholesterolaemic adults (0.02 ± 0.03%CVCmax /%baseline ) compared to both normocholesterolaemic (0.09 ± 0.02%CVCmax /%baseline ; P = 0.024) and statin-treated (0.12 ± 0.05%CVCmax /%baseline ; P = 0.03) adults. Chronic statin treatment improves reflex cutaneous vasodilatation in formerly hypercholesterolaemic older adults by increasing end-organ responsiveness to sympathetic outflow during passive heat stress.


Effects of close-up dietary energy level and supplementing rumen-protected lysine on energy metabolites and milk production in transition cows.
The objective of this study was to investigate the effects of dietary energy levels and rumen-protected lysine supplementation on serum free fatty acid levels, β-hydroxybutyrate levels, dry matter (DM) intake, and milk production and composition. Treatments were arranged in a 2 × 2 factorial design with 2 dietary energy levels [high net energy for lactation (NEL) = 1.53 Mcal/kg of DM vs. low NEL = 1.37 Mcal/kg of DM; HE vs. LE) fed either with rumen-protected lysine (bypass lysine; 40 g/cow per day) or without rumen-protected lysine (control). Sixty-eight third-lactation Holstein dairy cows entering their fourth lactation were randomly allocated to 4 treatments groups: HE with bypass lysine, HE without bypass lysine, LE with bypass lysine, and LE without bypass lysine. Groups were balanced based upon their expected calving date, previous milk yields, and body condition score. All cows were fed the same diet (NEL = 1.34 Mcal/kg of DM) during the dry period prior to the trial. Rumen-protected lysine was top-dressed on a total mixed ration to deliver 9.68 g/d of metabolizable lysine to pre- and postpartum cows. After calving, all cows received the same TMR (1.69 Mcal/kg of DM). Blood samples were collected at -21, -14, -7, 0, 3, 7, 14, and 21 d relative to calving, and free fatty acids and β-hydroxybutyrate concentrations were measured. Amount of feed offered and orts were collected and measured for individual cows 4 d/wk. Milk samples were collected once per week following calving, and milk composition was analyzed. Feeding high NEL to close-up cows decreased the concentrations of free fatty acid and β-hydroxybutyrate in prepartum cows but not in postpartum cows. Addition of rumen-protected lysine increased postpartum DM intake, and decreased serum free fatty acid and β-hydroxybutyrate concentrations. Neither energy nor rumen-protected lysine supplementation nor their interaction affected milk yield or fat or lactose yields. However, cows in the group receiving HE with bypass lysine tended to produce more milk compared with other groups and had a lower blood β-hydroxybutyrate concentration postpartum. These results indicate that feeding a high-energy diet together with rumen-protected lysine improved DM intake and lowered serum free fatty acid and β-hydroxybutyrate concentrations in transition cows.


Hole injection from P1 dye hot-excited states in p-type dye-sensitized films: a fluorescence study.
We present a study of the excited state relaxation dynamics of the photosensitizer P1 used in p-type dye-sensitized solar cells. Comparative femtosecond fluorescence upconversion measurements in solution and in films show that the dye undergoes a picosecond electronic relaxation from the bright Franck-Condon (FC) state to a low-emitting charge-transfer (CT) state in polar environment. The fluorescence is moderately quenched in solution and on the mesoporous Al2O3 isolator but dramatically more on NiO semiconductor. We assign this sub-picosecond process to the hole injection thus confirming that the electron transfer is from the FC state directly into the NiO valence band.


Genome-wide study of the defective sucrose fermenter strain of Vibrio cholerae from the Latin American cholera epidemic.
The 7th cholera pandemic reached Latin America in 1991, spreading from Peru to virtually all Latin American countries. During the late epidemic period, a strain that failed to ferment sucrose dominated cholera outbreaks in the Northern Brazilian Amazon region. In order to understand the genomic characteristics and the determinants of this altered sucrose fermenting phenotype, the genome of the strain IEC224 was sequenced. This paper reports a broad genomic study of this strain, showing its correlation with the major epidemic lineage. The potentially mobile genomic regions are shown to possess GC content deviation, and harbor the main V. cholera virulence genes. A novel bioinformatic approach was applied in order to identify the putative functions of hypothetical proteins, and was compared with the automatic annotation by RAST. The genome of a large bacteriophage was found to be integrated to the IEC224's alanine aminopeptidase gene. The presence of this phage is shown to be a common characteristic of the El Tor strains from the Latin American epidemic, as well as its putative ancestor from Angola. The defective sucrose fermenting phenotype is shown to be due to a single nucleotide insertion in the V. cholerae sucrose-specific transportation gene. This frame-shift mutation truncated a membrane protein, altering its structural pore-like conformation. Further, the identification of a common bacteriophage reinforces both the monophyletic and African-Origin hypotheses for the main causative agent of the 1991 Latin America cholera epidemics.


Walking for transportation in large Latin American cities: walking-only trips and total walking events and their sociodemographic correlates.
Walking for transportation is a common and accessible means of achieving recommended physical activity levels, while providing important social and environmental co-benefits. Even though walking in rapidly growing urban areas has become especially challenging given the increasing dependence on motorised transportation, walking remains a major mode of transportation in Latin American cities. In this paper we aimed to quantify self-reported walking for transportation in Mexico City, Bogota, Santiago de Chile, Sao Paulo, and Buenos Aires, by identifying both walking trips that are conducted entirely on foot and walking events involved in trips mainly conducted on other means of transportation (e.g. private vehicle, public transit) among individuals ≥5-years old. We show how walking-only trips account for approximately 30% trips in the analysed cities, and we evidence how the pedestrian dimension of mobility is largely underestimated if walking that is incidental to other transportation modes is not accounted for: when considering all walking events, we observed an increase between 73% and 217% in daily walking time. As a result, we estimated that between 19% and 25% of residents in these cities meet the WHO physical activity guidelines solely from walking for transportation. The results of the study also suggest that the promotion of public transportation in large Latin American cities can especially help certain population groups achieve the daily recommended levels of physical activity, while among low-income groups accessibility and safety seem to be the key challenges to be addressed.


Single-Stage Autologous Chondrocyte-Based Treatment for the Repair of Knee Cartilage Lesions: Two-Year Follow-up of a Prospective Single-Arm Multicenter Study.
There is an unmet need for a single-stage cartilage repair treatment that is cost-effective and chondrocyte-based. To evaluate the safety and preliminary efficacy of autologous freshly isolated primary chondrocytes and bone marrow mononucleated cells (MNCs) seeded into a PolyActive scaffold in patients with symptomatic cartilage lesions of the knee. Case series; Level of evidence, 4. A total of 40 patients with symptomatic knee cartilage lesions were treated with freshly isolated autologous chondrocytes combined with bone marrow MNCs delivered in a biodegradable load-bearing scaffold. The treatment requires only 1 surgical intervention and is potentially a cost-effective alternative to autologous chondrocyte implantation. The primary chondrocytes and bone marrow MNCs were isolated, washed, counted, mixed, and seeded into a load-bearing scaffold in the operating room. Patients were followed up at 3, 6, 12, 18, and 24 months. Primary endpoints were treatment-related adverse events up to 3 months, adverse implant effects between 3 and 24 months, and the implant success rate at 3 months as measured by lesion filling. Successful lesion filling (≥67% on magnetic resonance imaging) was found in 40 patients at 3 months and in 32 of the 32 patients analyzed at 24 months. Significant improvement over baseline was found for visual analog scale for pain from 3 months onward; Knee injury and Osteoarthritis Outcome Score (KOOS)-Pain and KOOS-Activities of Daily Living from 6 months onward; for KOOS-Symptoms and Stiffness, KOOS-Quality of Life and International Knee Documentation Committee from 12 months onward; and for KOOS-Sport and Recreation from 18 months onward. Hyaline-like repair tissue was found in 22 of 31 patients available for biopsy. Arthralgia and joint effusion were the most common adverse events. Scaffold delamination and adhesions led to removal of the implant in 2 patients. The treatment of knee cartilage lesions with autologous primary chondrocytes and bone marrow MNCs, both isolated and seeded into a load-bearing PolyActive scaffold within a single surgical intervention, is safe and clinically effective. Good lesion fill and sustained clinically important and statistically significant improvement in all patient-reported outcome scores were found throughout the 24-month study. Hyaline-like cartilage was observed on biopsy specimen in at least 22 of the 40 patients. NCT01041885 (ClinicalTrials.gov identifier).


Nummular headache: a series with symptomatic and primary cases.
Nummular headache (NH) is a primary headache adopting the form of local pain in a circumscribed area of < 7 cm in diameter in the tuber parietale, albeit it may also be located in other areas of the head. Although it is chronic, it is commonly associated with exacerbations and short periods of remission. Here we report four cases. Two of them could not be considered primary: in one the pain was related to an underlying, pointed and benign lesion disclosed only by magnetic resonance imaging (case 1); the second one had persistent NH days after trans-sphenoidal surgery for a pituitary adenoma, similar to a postcraniotomy headache (case 2). The two final patients suffered from typical forms of primary NH, one associated with migraine without aura, the other with chronic tension-type headache. The response to pain-related treatments and to preventive drugs was poor in the symptomatic as well as in the primary cases. The mechanisms are not clear, and peripheral (case 1) and also central pathways (case 2) could be involved. In the end, secondary forms of NH might coexist with classical primary NH. Particular attention should be paid to tiny skull lesions and to key events preceding the pain.


Studies on the Preparation, Characterization, and Solubility of 2-HP-β-Cyclodextrin-Meclizine HCl Inclusion Complexes.
Meclizine HCl is a poorly water-soluble drug having a very slow-onset of action. The effect of 2-hydroxypropyl-β-cyclodextrins and β-cyclodextrins on its aqueous solubility and dissolution rate was investigated. The phase solubility profile indicated that the solubility of Meclizine HCl was significantly increased in the presence of both 2-hydroxypropyl-β-cyclodextrin and β- cyclodextrin; an extend of increase being more for 2-hydroxypropyl-β-cyclodextrin. It was classified as AL-type, indicating the 1:1 stoichiometric inclusion complexes. The complexes formed were quite stable. The solid complexes prepared by physical mixtures, kneading methods, and co-precipitation methods were characterized using differential scanning calorimetry and FTIR. An in vitro study showed that the solubility and dissolution rate of Meclizine HCl were significantly improved by complexation with 2-hydroxypropyl-β-cyclodextrin. Tablet formulation using 1:1 kneading complex of Meclizine HCl and 2-hydroxypropyl-β-cyclodextrin with drug equivalent to 25 mg was prepared by a direct compression method. A dissolution study of prepared tablets was performed in 0.5% SLS in water (pH 7.0). Almost 96% drug was released from the formulation at the end of 30min. A comparison study of prepared tablets was done with marketed a Meclizine HCl 25 mg conventional tablet. From the results of dissolution study, it was found that the prepared formulation was showing better release, which was statistically significant P < 0.01 than a marketed tablet (paired t-test). Only 54% drug release was observed from the marketed tablet at the end of 30 min. Hence this study concludes that the solubility enhancement of Meclizine HCl could be successfully achieved using the inclusion complexation technique.


Professional identity formation of medical teachers in a non-Western setting.
Understanding and supporting professional identity formation (PIF) among medical teachers has become increasingly important in faculty development programs. In this study, we explored medical teachers' PIF in Indonesia. We conducted a qualitative descriptive study using focus group discussions (FGDs) at four medical schools in Indonesia. Basic science and clinical teachers from four different schools were selected to participate. Data were transcribed, coded, and analysed to develop themes and subthemes. Seventeen FGDs were completed, involving 60 basic science and 59 clinical teachers. Four major themes regarding the formation of medical teachers' professional identity emerged: an internal dialogue between intrinsic values and external influences, empowerment through early socialization, experiential workplace learning, and envisioning the future. The PIF process was similar for basic science and clinical teachers. Our findings suggested that PIF among medical teachers in a non-Western setting is a continuous and dynamic process that is shaped by key socialization factors (e.g. role models, workplace learning, peer support), with significant influences from religious beliefs, family values, and societal recognition. Faculty development programs should consider the dynamic and continuous nature of PIF among medical teachers and encourage clinicians and basic scientists to explore their values and beliefs, realize their goals, and envision their future.


Single particle inductively coupled plasma-mass spectrometry: a performance evaluation and method comparison in the determination of nanoparticle size.
Sizing engineered nanoparticles in simple, laboratory systems is now a robust field of science; however, application of available techniques to more complex, natural systems is hindered by numerous challenges including low nanoparticle number concentrations, polydispersity from aggregation and/or dissolution, and interference from other incidental particulates. A new emerging technique, single particle inductively coupled plasma-mass spectrometry (spICPMS), has the potential to address many of these analytical challenges when sizing inorganic nanoparticles in environmental matrices. However, to date, there is little beyond the initial feasibility studies that investigates the performance characteristics and validation of spICPMS as a nanoparticle sizing technique. This study compares sizing of four silver nanoparticle dispersions (nominal diameters of 40, 60, 80, and 100 nm) by spICPMS to four established sizing techniques: dynamic light scattering, differential centrifugal sedimentation, nanoparticle tracking analysis, and TEM. Results show that spICPMS is able to size silver nanoparticles, across different sizes and particle number concentrations, with accuracy similar to the other commercially available techniques. Furthermore, a novel approach to evaluating particle coincidence is presented. In addition, spICPMS size measurements were successfully performed on nanoparticles suspended in algal growth media at low concentrations. Overall, while further development of the technique is needed, spICPMS yields important advantages over other techniques when sizing nanoparticles in environmentally relevant media.


Atrial fibrillation cryoablation is an effective day case treatment: the UK PolarX vs. Arctic Front Advance experience.
Pulmonary vein isolation (PVI) is the cornerstone of catheter ablation for atrial fibrillation (AF). There are limited data on the PolarX Cryoballoon. The study aimed to establish the safety, efficacy, and feasibility of same day discharge for Cryoballoon PVI. Multi-centre study across 12 centres. Procedural metrics, safety profile, and procedural efficacy of the PolarX Cryoballoon with the Arctic Front Advance (AFA) Cryoballoon were compared in a cohort large enough to provide definitive comparative data. A total of 1688 patients underwent PVI with cryoablation (50% PolarX and 50% AFA). Successful PVI was achieved with 1677 (99.3%) patients with 97.2% (n = 1641) performed as day case procedures with a complication rate of <1%. Safety, procedural metrics, and efficacy of the PolarX Cryoballoon were comparable with the AFA cohort. The PolarX Cryoballoon demonstrated a nadir temperature of -54.6 ± 7.6°C, temperature at 30 s of -38.6 ± 7.2°C, time to -40°C of 34.1 ± 13.7 s, and time to isolation of 49.8 ± 33.2 s. Independent predictors for achieving PVI included time to reach -40°C [odds ratio (OR) 1.34; P < 0.001] and nadir temperature (OR 1.24; P < 0.001) with an optimal cut-off of ≤34 s [area under the curve (AUC) 0.73; P < 0.001] and nadir temperature of ≤-54.0°C (AUC 0.71; P < 0.001), respectively. This large-scale UK multi-centre study has shown that Cryoballoon PVI is a safe, effective day case procedure. PVI using the PolarX Cryoballoon was similarly safe and effective as the AFA Cryoballoon. The cryoablation metrics achieved with the PolarX Cryoballoon were different to that reported with the AFA Cryoballoon. Modified cryoablation targets are required when utilizing the PolarX Cryoballoon.


The Importance of Team Sex Composition in Team-Training Research Employing Complex Psychomotor Tasks.
The relationship between team sex composition and team performance on a complex psychomotor task was examined because these types of tasks are commonly used in the lab-based teams literature. Despite well-documented sex-based differences on complex psychomotor tasks, the preponderance of studies-mainly lab based-that use these tasks makes no mention of the sex composition of teams across or within experimental conditions. A sample of 123 four-person teams with varying team sex composition learned and performed a complex psychomotor task, Steal Beasts Pro PE. Each team completed a 5-hr protocol whereby they conducted several performance missions. The results indicated significant large mean differences such that teams with larger proportions of males had higher performance scores. These findings demonstrate the potential effect of team sex composition on the validity of studies that use complex psychomotor tasks to explore and investigate team performance-related phenomena when (a) team sex composition is not a focal variable of interest and (b) it is not accounted for or controlled. Given the proclivity of complex psychomotor action-based tasks used in lab-based team studies, it is important to understand and control for the impact of team sex composition on team performance. When team sex composition is not controlled for, either methodologically or statistically, it may affect the validity of the results in teams studies using these types of tasks.


Nek2-targeted ASO or siRNA pretreatment enhances anticancer drug sensitivity in triple‑negative breast cancer cells.
Although the anticancer drugs paclitaxel and doxorubicin are commonly used to treat many solid tumors, their effectiveness is highly variable due to tumor cell resistance. Therefore, it is important to find mechanisms that can be targeted to increase the sensitivity of cancer cells to current chemotherapeutic agents. NIMA‑related kinase 2 (Nek2), a serine/threonine kinase is emerging as an important oncogene because of its regulatory role in mitosis. Thus, regulation of the Nek2 expression levels may prove important as a target for cancer treatment. The purpose of our study was to determine whether drug sensitivity was increased in the triple negative breast cancer cell lines MDA-MB-231 and MDA-MB-468 by using small interfering RNA (siRNA) and antisense oligonucleotides (ASOs) against Nek2. To this end, MDA-MB-231 and MDA-MB-468 breast cancer cells transfected with Nek2 siRNA or ASO were exposed to various concentrations of paclitaxel and doxorubicin. Cell viability, cell cycle distribution and apoptosis were evaluated. We observed that drug susceptibility in these transfected cells was dramatically increased compared with either agent alone. FACS results showed that apoptosis was induced in siRNA- and ASO‑transfected cells as expected due to the regulatory function of Nek2 in centrosome duplication. Interestingly, the cell cyle was not arrested in transfected cells. We found that siRNA and ASO against Nek2 worked synergistically with paclitaxel and doxorubicin by promoting cell apoptosis. Our results suggest that these drugs in combination with Nek2 siRNA or ASO treatment may improve the sensitivity of cancer cells during chemotherapy treatments.


Loss of FHIT expression in breast cancer is correlated with poor prognostic markers.
The fragile histidine triad (FHIT) gene is a putative tumor suppressor gene that is thought to be involved in the carcinogenesis of breast cancer. Loss of FHIT expression has been observed in up to 72% of breast cancers and has been associated with increased p53, a high proliferation index, and increased tumor size and grade. However, loss of FHIT expression has not been investigated in association with apoptosis and cyclooxygenase-2 (COX-2) expression in breast cancer. Furthermore, expression of FHIT in primary breast tumors and their metastatic axillary lymph nodes has also not been previously described. The purpose of this study was to evaluate the expression of FHIT, COX-2, bcl-2, and p53 in primary breast tumor tissue; correlate their expression with known clinical and pathologic markers; and in cases when tissue was available, evaluate the expression of FHIT and COX-2 in the corresponding metastatic axillary lymph node in the same patient. Primary breast tumor specimens from 80 patients were examined for the presence of FHIT, COX-2, bcl-2, and p53 expression by immunohistochemistry using standard methods. When tissue was available, the expression of FHIT and COX-2 was also evaluated in the corresponding metastatic axillary lymph node specimen. FHIT expression in primary breast tumors was 56%. There was a significant correlation between FHIT expression in primary breast tumor and bcl-2 expression (P = 0.017). We also observed a significant inverse correlation between FHIT expression in primary breast tumor tissue and p53 expression (P = 0.023) in lymph node-negative cases. A significant inverse correlation between FHIT expression in the primary tumor and Ki-67 (P = 0.009) was also observed in lymph node-negative cases. FHIT expression in primary tumors correlated with FHIT expression in the metastatic lymph node (52.5%; P = 0.001). FHIT expression in primary tumors did not correlate with COX-2 expression. Our results suggest that loss of FHIT expression in breast cancer is associated with poor prognostic features. Furthermore, loss of FHIT expression is also seen in metastatic axillary lymph node. The prognostic and predictive value of these findings needs to be further evaluated in larger trials with longer follow-up.


Of liver, whisky and plants: a requiem for colchicine in alcoholic cirrhosis?
Colchicine decreases liver fibrosis in experimental and human disease, but a meta-analysis recently concluded that colchicine should not be used for liver fibrosis or cirrhosis irrespective of the aetiology. In this issue, Cortez-Pinto et al. confirm such negative conclusions in their series of 55 outpatients with biopsy-proven alcoholic cirrhosis followed for a median of 3.5 years. Although well tolerated, colchicine did not affect either the annual incidence rate of complications or liver function tests. Current treatment of alcoholic cirrhosis includes correction of nutritional deficiencies, exogenous administration of antioxidants (notably S-adenosylmethionine and polyenylphosphatidylcholine), and liver transplantation. In the future, preventive/therapeutic strategies will include campaigns to decrease alcohol abuse aimed at subjects genetically prone to develop alcoholic liver injury, prevention of liver fibrosis via inhibition of the Na+/H+ exchange, stimulation of apoptosis of stellate cells, antagonism of cytokines involved in liver injury, degradation of extracellular matrix, and reversal of ethanol-induced inflammatory and fibrotic changes via increased nitric oxide levels. On the grounds that it renders the hepatocyte more vulnerable to necrosis, steatosis has a key role in the pathogenesis of alcoholic and non-alcoholic liver disease. Conditions associated with insulin resistance have been recognized as risk factors for chronic liver disease and hepatocellular carcinoma in the alcoholic. This suggests that, through steatosis, insulin resistance could be a co-factor of alcoholic liver disease. Were such a hypothesis confirmed, it would unify our view of the pathogenesis of alcoholic and non-alcoholic liver disease, with all its inherent therapeutic implications.


Debrisoquine hydroxylase (CYP2D6) and prostate cancer.
The p450 hepatic microsomal enzyme system metabolizes exogenous drugs and carcinogens. Debrisoquine hydroxylase (CYP2D6), one member of the p450 hemoproteins, has polymorphic expression leading to poor metabolism of debrisoquine and similar compounds in approximately 7% of Caucasians. The genetic locus for this enzyme has been characterized, and the mutations responsible for the slowed metabolism have been identified. Epidemiological studies of the CYP2D6 phenotype suggest an association between the normal or rapid metabolism phenotype and increased risk of lung and bladder cancer. Preliminary data have also suggested an association with prostate cancer (CaP). We used a PCR-based assay to investigate possible associations between the CYP2D6 B allele, the most common genetic mutation responsible for the poor metabolism phenotype, and CaP. Using genomic DNA isolated from peripheral blood, we genetically typed 571 men with CaP and 767 matched controls, all participants in the Physician's Health Study. Relative to men homozygous for the wild-type allele, heterozygotes for the B allele have an odds ratio of 1.19 (95% confidence interval, 0.94-1.51) for CaP, and men homozygous for the B allele have an odds ratio of 1.37 (95% confidence interval, 0.86-2.20). When analyzed as a trend over zero, one, or two copies of the B allele, there emerges a possible association between the B allele and an increased risk of CaP of borderline statistical significance (P = 0.07).


Hierarchical shape description via the multiresolution symmetric axis transform.
A method is proposed that produces a shape description in the form of a hierarchy by scale of simple symmetric axis sequences. An axis segment that is a child of another has smaller scale and is seen as a branch of its parent. The scale value and parent-child relationship are induced by following the symmetric axis under successive reduction of resolution. The result, in two or three dimensions, is a figure¿rather than boundary¿oriented shape description that has natural segments and is insensitive to noise in the object description. The general approach of hierarchy production by following a feature through successive resolution reduction will be presented, as will methods of resolution reduction and computer implementation. Also, the relation of this figure-based shape description to those based on boundary curvature will be briefly discussed.


What are the levels of evidence on which we base decisions for surgical management of lower extremity bone tumors?
Benign and malignant lower extremity primary bone tumors are among the least common conditions treated by orthopaedic surgeons. The literature supporting their surgical management has historically been in the form of observational studies rather than prospective controlled studies. Observational studies are prone to confounding bias, sampling bias, and recall bias. (1) What are the overall levels of evidence of articles published on the surgical management of lower extremity bone tumors? (2) What is the overall quality of reporting of studies in this field based on the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist? (3) What are the most common pitfalls in reporting that authors might improve on? All studies describing the surgical management of lower extremity primary bone tumors from 2002 to 2012 were systematically reviewed. Two authors independently appraised levels of evidence. Quality of reporting was assessed with the STROBE checklist. Pitfalls in reporting were quantified by determining the 10 most underreported elements of research study design in the group of studies analyzed, again using the STROBE checklist as the reference standard. Of 1387 studies identified, 607 met eligibility criteria. There were no Level I studies, two Level II studies, 47 Level III studies, 308 Level IV studies, and 250 Level V studies. The mean percentage of STROBE points reported satisfactorily in each article as graded by the two reviewers was 53% (95% confidence interval, 42%-63%). The most common pitfalls in reporting were failures to justify sample size (2.2% reported), examine sensitivity (2.2%), account for missing data (9.8%), and discuss sources of bias (14%). Followup (66%), precision of outcomes (64%), eligibility criteria (55%), and methodological limitations (53%) were variably reported. Observational studies are the dominant evidence for the surgical management of primary lower extremity bone tumors. Numerous deficiencies in reporting limit their clinical use. Authors may use these results to inform future work and improve reporting in observational studies, and treating surgeons should be aware of these limitations when choosing among the various options with their patients.


PARP Inhibitor Inhibits the Vasculogenic Mimicry through a NF-κB-PTX3 Axis Signaling in Breast Cancer Cells.
Poly (ADP-ribose) polymerase inhibitors (PARPi) are targeted therapies that inhibit PARP proteins which are involved in a variety of cell functions. PARPi may act as modulators of angiogenesis; however, the relationship between PARPi and the vasculogenic mimicry (VM) in breast cancer remains unclear. To determine whether PARPi regulate the vascular channel formation, we assessed whether the treatment with olaparib, talazoparib and veliparib inhibits the vascular channel formation by breast cancer cell lines. Here, we found that PARPi act as potent inhibitors of the VM formation in triple negative breast cancer cells, independently of the BRCA status. Mechanistically, we find that PARPi trigger and inhibit the NF-κB signaling, leading to the inhibition of the VM. We further show that PARPi decrease the expression of the angiogenic factor PTX3. Moreover, PTX3 rescued the PARPi-inhibited VM inhibition. In conclusion, our results indicate that PARPi, by targeting the VM, may provide a new therapeutic approach for triple negative breast cancer.


Vitamin K intake and the risk of fractures: A meta-analysis.
The association between dietary vitamin K intake and the risk of fractures is controversial. Therefore we perform a meta-analysis of cohort or nested case-control studies to investigate the relationship between dietary vitamin K intake and the risk of fractures. A comprehensive search of PubMed and EMBASE (to July 11, 2016) was performed to identify cohort or nested case-control studies providing quantitative estimates between dietary vitamin K intake and the risk of fractures. Summary relative risk (RRs) with corresponding 95% confidence intervals (CIs) were pooled by using a random-effects model. Four cohort studies and one nested case-control study, with a total of 1114 fractures cases and 80,982 participants, were included in our meta-analysis. Vitamin K intake in all included studies refers exclusively to the intake of phylloquinone (vitamin K1), which is the predominant form of vitamin K in foods. We observed a statistically significant inverse association between dietary vitamin K intake and risk of fractures (highest vs. the lowest intake, RR = 0.78, 95% CI: 0.56-0.99; I = 59.2%, P for heterogeneity = .04). Dose-response analysis indicated that the pooled RR of fracture for an increase of 50 μg dietary vitamin K intake per day was 0.97 (95% CI: 0.95-0.99) without heterogeneity among studies (I = 25.9%, P for heterogeneity = .25). When stratified by follow-up duration, the RR of fracture for dietary vitamin K intake was 0.76 (95% CI: 0.58-0.93) in studies with more than 10 years of follow-up. Our study suggests that higher dietary vitamin K intake may moderately decrease the risk of fractures.


Ki-67 immunostaining in node-negative stage I/II breast carcinoma. Significant correlation with prognosis.
Prognostic predictors for node-negative breast carcinoma have not been clearly established. Immunostaining with Ki-67 antibody was performed on frozen sections of histologically proved node-negative breast carcinomas from 42 patients to examine its prognostic value and its association with other clinicopathologic and biochemical parameters, i.e., patient age and tumor size, histologic type, nuclear grade, mitotic rate, presence of vascular or lymphatic invasion, DNA ploidy, percentage of cells in S-phase, estrogen content, and c-erbB-2 amplification. Thirty-seven of the 42 tumors showed immunoreactivity with Ki-67 antibody in 1% to 55% of the tumor cells. A strongly significant correlation was observed between Ki-67 staining percentage and, respectively, nuclear grade, age, and mitotic rate. Nuclear grade 1 (the most anaplastic) tumors showed a significantly higher median percentage of cells stained (median, 14; range, 3 to 40) compared with nuclear grade 3 tumors (median, 0.5; range, 0 to 8). Thirteen patients developed recurrence; six of them died of disease. On univariate analysis, both 5-year disease-free and overall survivals were strongly associated with percentage of cells stained with Ki-67 antibody. Our results suggest that Ki-67 immunostaining correlates well with nuclear grade and clinical outcome in node-negative breast carcinoma. Because of small sample size analyzed in this study we were unable to do multivariate analysis. Therefore, further studies with larger number of cases are needed to determine whether tumor proliferative activity determined by Ki-67 immunostaining is an independent prognostic parameter or it merely reflects histopathologic features such as nuclear grade or mitotic activity.


The electrophysiology of the olfactory-hippocampal circuit in the isolated and perfused adult mammalian brain in vitro.
The viability and general electrophysiological properties of the limbic system in the adult mammalian brain isolated and maintained in vitro by arterial perfusion are described. The isolated brain preparation combines the advantages of intact synaptic connectivity and accessibility of different areas of the encephalic mass with those of the in vitro approach, i.e., stability and control of the ionic environment. Extracellular field potential as well as intracellular recordings were performed at different levels in the limbic system of isolated adult guinea pig brains. The results demonstrate that in the piriform, entorhinal, and hippocampal cortices, the intrinsic electrical properties of individual cells as well as the spontaneous and evoked electrical activity in the neuronal ensembles they comprise, were virtually identical to those observed in vivo. The properties of the limbic system loop were determined.


New aqueous inflow inhibitors.
Several new aqueous inflow inhibitors have been introduced over the past few years. These new agents include two alpha agonists, two topical carbonic anhydrase inhibitors, and a novel combination agent. All of these agent compare favorably with timolol in terms of aqueous suppression, though each has its own relative advantages and disadvantages.


Is Brucella excreted in cattle faeces? - Evidence from Punjab, India.
Brucellosis is a neglected zoonosis that affects animals and people in much of the underdeveloped world. The disease is endemic in cattle in Punjab, India and controlling it is a public health challenge. Dairy farmers and farm labour commonly handle cattle faeces with bare hands and personal protective equipments are not used. No studies have been conducted about the shedding of Brucella species in faeces of sero positive cattle in the state. This study aimed to isolate and identify the Brucella species from faeces of sero positive cattle in Punjab, India. Faecal samples were collected from 350 Brucella sero positive cattle in Ludhiana district of Punjab, India. Isolation was performed using a pre-enriched Brucella selective broth medium as well as Brucella selective medium agar plates containing horse serum and Brucella selective supplements. Isolates were identified using Gram staining technique and rapid slide agglutination test, and then confirmed by using bcsp31 and 16s rRNA genus specific PCR. Isolates were further identified up to species level by using Bruce-Ladder multiplex PCR. Fourteen Brucella species were isolated, all of which showed coccobacilli on gram staining, positive rapid slide agglutination test and amplification of bcsp31 and 16s rRNA genes. Of the 14 isolates, 11 were identified as Brucella abortus and 3 were identified as Brucella melitensis. The study demonstrates that animal faeces could pose a potential risk for animal and human health and faeces of seropositive cattle must be handled with care.


Influence of nicotine on healing of vesical sutures in rabbits.
To study the effects of nicotine on inflammatory cells, deposition of collagen and its interference on the strength of tissue in vesical sutures in rabbits. Twenty New Zealand rabbits were used, randomized into two groups: group N, consisting of 10 animals, to which nicotine was administered in the dose of 2mg/kg of weight as a subcutaneous injection, diluted in 1 ml of saline solution at 0.9% in a daily administration during the 28 days prior to the surgery; and group C, consisting of 10 animals, to which saline solution at 0.9% was administered in the same conditions and time intervals of the nicotine group. All the animals underwent cystotomy and suture of the bladder wall 28 days after the administration of nicotine or saline solution. The measurements were performed on the fourth and seventh day in each group after cystectomy and euthanasia of the animals. A fragment of bladder was removed and sent for a tensile strength test to evaluate the tissue strength and another fragment underwent a histological analysis of inflammatory process and deposition of collagen. There was a decrease of neutrophils on the fourth postoperative day (p=0.079) and an increase of plasmocytes on the seventh postoperative day (p=0.053) in the animals that were given nicotine, without statistical difference in relation to the control group. In the analysis of the proliferation of fibroblasts, deposition of collagen and tensile strength test, there was no statistical difference in relation to the control group. The administration of nicotine in rabbits did not influence the healing process of vesical suture in relation to the inflammatory cells, deposit of collagen and tissue strength of the suture.


BicPAM: Pattern-based biclustering for biomedical data analysis.
Biclustering, the discovery of sets of objects with a coherent pattern across a subset of conditions, is a critical task to study a wide-set of biomedical problems, where molecular units or patients are meaningfully related with a set of properties. The challenging combinatorial nature of this task led to the development of approaches with restrictions on the allowed type, number and quality of biclusters. Contrasting, recent biclustering approaches relying on pattern mining methods can exhaustively discover flexible structures of robust biclusters. However, these approaches are only prepared to discover constant biclusters and their underlying contributions remain dispersed. The proposed BicPAM biclustering approach integrates existing principles made available by state-of-the-art pattern-based approaches with two new contributions. First, BicPAM is the first efficient attempt to exhaustively mine non-constant types of biclusters, including additive and multiplicative coherencies in the presence or absence of symmetries. Second, BicPAM provides strategies to effectively compose different biclustering structures and to handle arbitrary levels of noise inherent to data and with discretization procedures. Results show BicPAM's superiority against its peers and its ability to retrieve unique types of biclusters of interest, to efficiently deliver exhaustive solutions and to successfully recover planted biclusters in datasets with varying levels of missing values and noise. Its application over gene expression data leads to unique solutions with heightened biological relevance. BicPAM approaches integrate existing disperse efforts towards pattern-based biclustering and provides the first critical strategies to efficiently discover exhaustive solutions of biclusters with shifting, scaling and symmetric assumptions with varying quality and underlying structures. Additionally, BicPAM dynamically adapts its behavior to mine data with different levels of missing values and noise.


Incidence and associated risk factors of antituberculosis drug-induced hepatotoxicity among hospitalised patients in Wuhan, China.
To analyse the incidence and risk factors of hepatotoxicity induced by antituberculosis (anti-TB) drugs in Renmin Hospital of Wuhan University, and to provide evidence for clinical prevention and treatment of anti-TB drug damage. A retrospective analysis of patients who received first-line anti-TB drugs from January 2016 to December 2018 in Renmin Hospital of Wuhan University was conducted. Univariate analysis and binary logistic regression analysis with the forward stepwise method were used to assess the risk factors associated with hepatotoxicity induced by anti-TB drugs. Of the 1603 patients treated with anti-TB drugs, only 1115 patients met the inclusion criteria and 42 subjects developed anti-TB drug-induced hepatotoxicity (ATDH). Significant differences (p<0.05) were seen in age (p=0.042), hypertension (p=0.021), treatment duration (p=0.000) and therapeutic regimen (p=0.001) between the non-ATDH and ATDH groups. Regression analysis further indicated that treatment duration (OR 1.053, 95% CI 1.031 to 1.076, p=0.000) and therapeutic regimens such as isoniazid (H), rifampicin (R), pyrazinamide (Z) and streptomycin (S) (HRZS) (OR 5.751, 95% CI 2.318 to 14.267, p=0.000), HRZ (OR 3.546, 95% CI 1.449 to 8.676, p=0.006) and RZ (OR 12.243, 95% CI 1.181 to 126.862, p=0.036) were risk factors for ATDH. The incidence of ATDH in this study was 3.77%, which was lower than that of other hospital-based studies. Treatment duration and therapeutic regimen might be potential risk factors for ATDH during anti-TB therapy in hospital. Among these therapeutic combination regimens, HRZS, HRZ and RZ could significantly increase the occurrence of ATDH when used as anti-TB therapy, while isoniazid, rifampicin and ethambutol (HRE) might be safer.


In vitro and in vivo relaxation of urinary bladder smooth muscle by the selective myosin II inhibitor, blebbistatin.
OBJECTIVE To investigate the in vitro and in vivo effects of blebbistatin (a small cell-permeable molecule with high affinity and selectivity toward the myosin II contractile molecule) on bladder smooth muscle (SM) contractility, as antimuscarinic therapy is only 65-75% effective in treating overactive bladder (OAB) and is associated with considerable side-effects, with a < 25% continuation rate at 1 year. MATERIALS AND METHODS Bladder and aortic strips from adult male rats, and human bladder strips obtained from open prostatectomy, were used for organ-bath studies of blebbistatin. Awake cystometry was also used in rats in both the presence and absence of intravesically delivered blebbistatin. RESULTS Blebbistatin dose-dependently and completely relaxed both KCl- and carbachol-induced rat detrusor and endothelin-1-induced human bladder contraction. Pre-incubation with 10 µm blebbistatin attenuated carbachol responsiveness by ≈ 65% while blocking electrical field stimulation-induced bladder contraction reaching 50% inhibition at 32 Hz. The basal tone and amplitude of spontaneous contraction were also significantly diminished. Urodynamic variables were obviously altered by intravesical infusion with blebbistatin. CONCLUSION Our novel data show that blebbistatin strongly relaxes both rat and human bladder contraction induced by various physiological stimuli. Coupled with our in vivo data showing that nanomole doses of blebbistatin significantly alter urodynamic variables to produce a less active bladder, our results suggest the possibility of intravesically administered blebbistatin binding at myosin II being developed as a therapeutic treatment for OAB via a novel targeting of the SM contractile apparatus.


Mitogen-initiated synthesis and secretion of T cell growth factor(s) by a T-lymphoma cell line.
Of 12 T-lymphoma cell lines investigated, one line, EL-4 azgr (Thy-1+, Lyt-1+, Lyt-2-3-,Ia-, T-200+, sIg-, and FcR-) and, to a lesser extent, the parental cell line, EL-4, produced T cell growth factor(s) (TCGF) when stimulated by the T-cell mitogen concanavalin A (Con A). Induced production of TCGF-E was detected by 6 h and maximal at 18-24 h. Purified TCGF-E from this source had an approximately 30,000 mol wt and the biological activity of TCGF produced by whole spleen cells, including: augmentation of T cell-mitogen responses, cytotoxic T lymphocyte (CTL) proliferation support dependence, augmented generation of CTL, lack of strain specificity, and failure to stimulate resting T cells. TCGF-E is neither synthesized or secreted by this lymphoma cell line unless stimulated by Con A. X-irradiation up to 7,000 rad failed to inhibit synthesis and secretion. These observations have a practical application in providing a relatively homogeneous clonal cell product for T cell culture support and for structural and functional studies of the TCGF molecule(s). They suggest also a model for examining mechanisms of triggering production and secretion of a regulatory molecule that controls T cell functions.


Credentialing and job practice in environmental health: an empirical study.
To investigate the validity of the credentialing examination for entry-level practitioners in environmental health, 15 work measures, simulating or assessing important components of job practice, were developed. These work measures, along with the written examination, were administered to a sample of 128 entry-level practitioners drawn from 10 test sites throughout the country. Composite work measures were constructed in which individual work measures were weighted according to their judged importance. The total composite work measure, based on all 15 individual measures, correlated .53 with the total written examination. Correlations between composite work measures and the written test scores were generally in the .3 to .5 range and significant at the .01 level. Seventy-three percent of the sample were consistently classified on both the written examination and the total work measure composite. That is, 73 percent of the sample passed both the examination and the work measures or failed both the examination and the work measures. In comparison to similar studies, the magnitude of the coefficients reported here is acceptable to high. Results show that the examination is a valid, but not perfect, predictor of on-the-job skill in environmental health, and it should help to screen the public from the results of incompetent practice.


Kinetics of the sodium pump in red cells of different temperature sensitivity.
Ouabain-sensitive K influx into ground squirrel and guinea pig red cells was measured at 5 and 37 degrees C as a function of external K and internal Na. In both species the external K affinity increases on cooling, being three- and fivefold higher in guinea pig and ground squirrel, respectively, at 5 than at 37 degrees C. Internal Na affinity also increased on cooling, by about the same extent. The effect of internal Na on ouabain-sensitive K influx in guinea pig cells fits a cubic Michaelis-Menten-type equation, but in ground squirrel cells this was true only at high [Na]i. There was still significant ouabain-sensitive K influx at low [Na]i. Ouabain-binding experiments indicated around 800 sites/cell for guinea pig and Columbian ground squirrel erythrocytes, and 280 sites/cell for thirteen-lined ground squirrel cells. There was no significant difference in ouabain bound per cell at 37 and 5 degrees C. Calculated turnover numbers for Columbian and thirteen-lined ground squirrel and guinea pig red cell sodium pumps at 37 degrees C were about equal, being 77-100 and 100-129 s-1, respectively. At 5 degrees C red cells from ground squirrels performed significantly better, the turnover numbers being 1.0-2.3 s-1 compared with 0.42-0.47 s-1 for erythrocytes of guinea pig. The results do not accord with a hypothesis that cold-sensitive Na pumps are blocked in one predominant form.


KRAS Mutation Detection in Non-small Cell Lung Cancer Using a Peptide Nucleic Acid-Mediated Polymerase Chain Reaction Clamping Method and Comparative Validation with Next-Generation Sequencing.
KRAS is one of commonly mutated genetic "drivers" in non-small cell lung cancers (NSCLCs). Recent studies indicate that patients with KRAS-mutated tumors do not benefit from adjuvant chemotherapy, so there is now a focus on targeting KRAS-mutated NSCLCs. A feasible mutation detection method is required in order to accurately test for KRAS status. We compared direct Sanger sequencing and the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method in 134 NSCLCs and explored associations with clinicopathological factors. Next-generation sequencing (NGS) was used to validate the results of discordant cases. To increase the resolution of low-level somatic mutant molecules, PNA-mediated PCR clamping was used for mutant enrichment prior to NGS. Twenty-one (15.7%) cases were found to have the KRAS mutations using direct sequencing, with two additional cases by the PNA-mediated PCR clamping method. The frequencies of KRAS mutant alleles were 2% and 4%, respectively, using conventional NGS, increasing up to 90% and 89%, using mutant-enriched NGS. The KRAS mutation occurs more frequently in the tumors of smokers (p=.012) and in stage IV tumors (p=.032). Direct sequencing can accurately detect mutations, but, it is not always possible to obtain a tumor sample with sufficient volume. The PNA-mediated PCR clamping can rapidly provide results with sufficient sensitivity.


Urinary retention without organic cause in children.
Acute urinary retention without organic explanation is well known in adults but has only rarely been described in children. Six cases (four boys and two girls) of non-organic acute urinary retention are reported. The diagnosis was based on negative findings on extensive urological examination. So-called psychogenic urinary retention in children is associated with constipation and infrequent voiding.


Sarcocystis camelicanis increases interleukin (IL)-6 expression in one-humped camels (Camelus dromedarius) from Riyadh and Al Qassim, Saudi Arabia.
Sarcocystis spp. are intracellular protozoan parasites with an intermediate-definitive host life cycle based on a prey-predator relationship. Sarcocystis infection is common among different vertebrates including humans. The pathogenicity of Sarcocystis spp. is of varied significance including a possible lethal effect for the host. The goal of the present study was to investigate the inflammatory activity of Sarcocystis spp. in different organs of naturally infected camels. The tongue, esophagus, heart, diaphragm, and skeletal muscles were collected from 50 camels, and the tissues assessed for the presence of Sarcocystis spp. by macroscopic examination, light microscopy, and transmission electron microscopy (TEM). Moreover, expression of the interleukin (IL)-6 was analyzed using reverse transcriptase quantitative polymerase chain reaction (qPCR). Microscopic Sarcocystis spp. cysts were found in camels. TEM identified the cysts as Sarcocystis camelicanis (S. camelicanis). Sarcocystis infection increased inflammation by stimulation of IL-6 expression in different organs of the camels, particularly in those from the Al-Qassim region.


Trends in antidepressant prescribing for new episodes of depression and implications for health system quality measures.
The nationally reported Healthcare Effectiveness Data and Information Set (HEDIS) antidepressant medication management measure assesses whether patients with new episodes of depression receive antidepressant coverage for 84 of the first 114 days of treatment. Although initial prescriptions for a 90-day supply satisfy measure requirements, they may circumvent its purpose of ensuring adequate medication management. To assess the extent to which 90-day initial prescriptions have contributed to health system performance on the HEDIS antidepressant measure from fiscal years 2001 to 2008. Retrospective cohort analysis of Veterans Health Administration administrative data. Patients with a new diagnosis of depression and a new antidepressant prescription (N=383,634). HEDIS antidepressant measures, days supply of initial antidepressant prescriptions, antidepressant refills, and clinical encounters. Health system performance on the HEDIS acute phase antidepressant measure increased from 63.1% in 2001 to 71.0% in 2008. Receipt of an initial 90-day antidepressant supply increased from 10.5% to 29.1% during this same period; when these are excluded, HEDIS performance was 58.8% in 2001 and 59.4% in 2008. Receiving an initial 90-day prescription was associated with prior antidepressant treatment, fewer clinical encounters, and similar rates of antidepressant refills compared with patients prescribed less than 90-day supplies. Although increases in initial 90-day supplies contribute to improved performance on the HEDIS measure, actual adherence during the acute treatment phase may not be changed by this practice. Quality measures based on pharmacy fills may need modification in the setting of large initial prescriptions.


Long-term safety and efficacy study of adapalene 0.3% gel.
The efficacy and safety of adapalene 0.1% gel in the treatment of acne vulgaris has been demonstrated in multiple controlled clinical trials. A higher concentration formulation, adapalene 0.3% gel, has been developed to provide a broader range of treatment options for acne management. Phase 3 clinical studies have demonstrated the superior efficacy of adapalene 0.3% gel compared to adapalene 0.1% gel and its vehicle at the end of a 12-week treatment period. The goal of this study was to evaluate the long-term safety of adapalene 0.3% gel in subjects treated once daily for 52 weeks, with a secondary objective to evaluate long-term efficacy. Subjects 12 years of age or older (N=551) with acne vulgaris participated in a multicenter, open-label study of the long-term (up to 52 weeks) efficacy and safety of once-daily applications of adapalene 0.3% gel. Of those enrolled, 167 subjects completed 12 months of treatment. Expected signs and symptoms of local cutaneous irritation (erythema, dryness, scaling, and stinging/burning) were mostly mild or moderate, with mean tolerability scores below 1 (mild) at all time points for the parameters assessed. Treatment-related, dermatologic adverse events were experienced by 21% of subjects and dry skin, skin discomfort, and scaling were reported by 10.5%, 8.3% and 3.3% of subjects, respectively. Most of the adverse events reported occurred in the first quarter of treatment. Adverse events were mostly mild to moderate in severity. Subjects treated with adapalene 0.3% gel for 52 weeks achieved a >75% median reduction in total, inflammatory, and noninflammatory lesions in this open-label study by the end of the treatment period. Adapalene 0.3% gel was safe and effective in the long-term (up to 1 year) treatment of subjects with acne vulgaris.


[Testing for cannabis and synthetic cannabinoids in human specimens].
Among the 60 or so various cannabinoids which are present in cannabis sativa indica, terpenophenols are mainly found, including delta-9-trans tetrahydrocannabinol (Δ9-THC) which is the major psychoactive ingredient. Over the last decade, due to the emergence of e-commerce and the continuous investigations by pharmaceutical groups to identify new active molecules, synthetic cannabinoids have been proposed. These compounds, under the generic name "spices" have chemical structures very different from that of Δ9-THC, but share the same CB1 and CB2 receptors. They are mimics of Δ9-THC with much powerful pharmacological effects, including thus more deterrent side-effects, and enhanced windows of detection. All these drugs are controlled. This means that they have to be tested in human biological specimens to document abuse. The aim of this mini-review is to present what has been described in the scientific literature according to the available specimens (blood, urine, saliva, sweat, hair and exhaled breath), focussing on the current advantages and limitations of each test.


The Effects of Cognitive Control and Time on Frontal Beta Oscillations.
Frontal beta oscillations are associated with top-down control mechanisms but also change over time during a task. It is unclear whether change over time represents another control function or a neural instantiation of vigilance decrements over time, the time-on-task effect. We investigated how frontal beta oscillations are modulated by cognitive control and time. We used frontal chronic electrocorticography in monkeys performing a trial-and-error task, comprising search and repetition phases. Specific beta oscillations in the delay period of each trial were modulated by task phase and adaptation to feedback. Beta oscillations in this same period showed a significant within-session change. These separate modulations of beta oscillations did not interact. Crucially, and in contrast to previous investigations, we examined modulations of beta around spontaneous pauses in work. After pauses, the beta power modulation was reset and the cognitive control effect was maintained. Cognitive performance was also maintained whereas behavioral signs of fatigue continued to increase. We propose that these beta oscillations reflect multiple factors contributing to the regulation of cognitive control. Due to the effect of pauses, the time-sensitive factor cannot be a neural correlate of time-on-task but may reflect attentional effort.


Bimanual laryngoscopy: a videographic study of external laryngeal manipulation by novice intubators.
We perform a videographic analysis of external laryngeal manipulation (ELM) by novice intubators using a direct laryngoscopic imaging system and a validated means of assessing laryngeal view (percentage of glottic opening [POGO] score). Nine first-year emergency medicine residents performed a total of 484 laryngoscopies during a 1-year study period, of which 271 were videotaped. Of this convenience sample, cases were included in the study if the initial laryngeal view had a POGO score of less than 50% and the quality of video imaging permitted POGO scoring before and after application of ELM. Seventy-two cases met the study criteria. Initial POGO scores equaled 0% in 30 cases, less than 20% in 20 cases, and 20% to 50% in 22 cases. ELM improved the POGO scores in all cases (95% confidence interval [CI] 96% to 100%) by an average of 57% (SD 23.7; 95% CI 40% to 75%). Among those with an initial POGO score equal to 0%, the mean improvement was 61% (SD 36.3; 95% CI 48% to 74%). For those with an initial POGO score of less than 20%, the mean improvement was 63% (SD 19.5; 95% CI 54% to 71%), and for those with an initial POGO score of 20% to 50%, the mean improvement was 47% (SD 16.0; 95% CI 40% to 55%). ELM is a simple and very effective technique for improving laryngeal view during laryngoscopy by novice intubators.


[The present and future of home care for patients with gastrointestinal malignant diseases].
Home care for patients with gastrointestinal malignancies includes nutritional support, chemotherapy and hospice care. Every step should be started after acquiring sufficient information on the disease. Home infusion therapy (HIT) is most important for nutritional care because the patient usually cannot eat or eats very little. It should be started fast within another symptoms controllable. With some problems in access procedure, total calories, formula, catheter infection, it is important to treat individually not to over dose. As for home chemotherapy, new molecular target-based therapies that prolong tumor dormancy are expected to be developed. Regarding home hospice, the control of pain, nausea, general fatigue, and other symptoms is the key to continuous home hospice care. The medical linkage between hospital and clinic, clinic and clinic and skillful visiting nurses is indispensable and the establishment and quality of medical systems, networks play an important role in the home care of patients of gastrointestinal malignancies.


H2O2 is an important mediator of UVB-induced EGF-receptor phosphorylation in cultured keratinocytes.
Exposure of human keratinocytes to physiologic doses of ultraviolet B (UVB) radiation induces phosphorylation of the epidermal growth factor receptor (EGFR). We demonstrate that H2O2 generated by UVB mediates EGFR phosphorylation. Using dihydrorhodamine 123 as a specific fluorescent dye probe, we show that UVB irradiation (50-800 J per m2) of keratinocytes leads within minutes to concentration-dependent intracellular production of H2O2. A corresponding concentration-dependent increase in the release of extracellular H2O2 was measured by using Amplex, a derivative of dihydrophenoxazine. The levels of intracellular H2O2 that are induced by UVB irradiation and that stimulate EGFR phosphorylation correlate strongly with the response induced by exogenously added H2O2. UVB or H2O2 demonstrated concentration- and time-dependent stimulation of EGFR phosphorylation that was initially observed within 1-5 min and exhibited a proportionate delay for UVB-induced production of H2O2. EGFR phosphorylation induced by UVB or H2O2 declined significantly toward baseline levels by 4 h and could be restimulated after H2O2 but not after UVB exposure. Phosphorylation of EGFR was inhibited by the structurally unrelated antioxidants butylated hydroxyanisole, N-acetyl-L-cysteine, and pyrrolidine dithiocarbamate, or by the H2O2-degrading enzyme catalase. These data indicate that generation of H2O2 by UVB radiation of human keratinocytes participates in the rapid, ligand-independent phosphorylation of EGFR and implicate H2O2 as a biologic mediator in EGFR activation and regulation of the downstream signaling cascade. UVB-induced H2O2 has the potential to initiate or modulate early EGFR-mediated signaling events that could play an important role in the cellular response to oxidative stress.


Eugenia uniflora Dentifrice for Treating Gingivitis in Children: Antibacterial Assay and Randomized Clinical Trial.
School-age children are frequently at high risk for the onset of biofilm-dependent conditions, including dental caries and periodontal diseases. The objective of this study was to evaluate the clinical efficacy of a dentifrice containing Eugenia uniflora Linn. (Surinam cherry) extract versus a triclosan-based comparator in treating gingivitis in children aged 10-12 years. The in vitro antibacterial potential of the dentifrice was tested against oral pathogens (Streptococcus mutans, Streptococcus oralis and Lactobacillus casei). Then a phase-II clinical trial was conducted with 50 subjects aged 10-12 years, with clinical signs of gingivitis. The subjects were randomly assigned to the experimental group (n=25) and control group (n=25), in which participants used the experimental dentifrice and a triclosan-based fluoridated dentifrice (Colgate Total 12(r)), respectively. Clinical examinations assessed the presence of gingivitis (primary outcome) and biofilm accumulation (secondary outcome) using the Gingival-Bleeding Index (GBI) and Simplified Oral Hygiene Index (OHI-S), respectively, at baseline and after seven days of tooth brushing 3x/day. The data were analyzed using paired and unpaired t-test (GBI) and Wilcoxon and Mann-Whitney (OHI-S), with p≤0.05. The experimental dentifrice showed efficient antibacterial activity in vitro. In the clinical trial, a significant reduction in gingival bleeding was observed in both experimental and control groups (p<0.0001), with no statistical difference between them (p=0.178), although a small size effect was observed. Biofilm accumulation was only reduced in the control group (p=0.0039). In conclusion, E. uniflora dentifrice showed anti-gingivitis properties in children aged 10-12 years. Thus, it may be a potentially efficient and safe product to be used alternatively in preventive dental practice.


Process evaluation of the New South Wales Walk Safely to School Day.
Concerns about the contribution of lower physical activity by children on levels of childhood obesity have driven the search for interventions that encourage lifestyle physical activity by school-aged children. This paper describes the evaluation of the New South Wales (NSW) Walk Safely To School Day (WSTSD), an event repeated annually on the first Friday in April from 2001 to 2004. Paid media advertising before the event promoted WSTSD. All primary schools in NSW were invited to participate. The evaluation included monitoring of schools registering to participate over four years, and in 2002 evaluation feedback from schools as well as a population-based telephone survey of randomly selected parents of primary school-aged children. By 2004, 53% of all primary schools in NSW had participated at least once in WSTSD and 15% had participated for at least three years. Participating schools linked WSTSD to road safety curricula, and 59% of schools also talked about WSTSD at school assemblies, but few (7%) organised walking-related activities. Parents reported that one-quarter (24%) of NSW primary school-aged students participated in WSTSD activities, including an additional 31% more children walking to school on WSTSD than would normally walk on a Friday. Walk Safely To School Day resulted in a moderate short-term change in walking behaviour. High levels of in-principle support from parents indicate that greater participation in walking to school is possible, but will require stronger interventions to overcome the barriers identified by parents and to maintain changes.


[An electrographic analysis of the dynamic change in the functional state of the rat brain after destruction of the locus coeruleus].
Bilateral destruction of the Locus coeruleus in rats was carried out by means of 6-hydroxydopamine injections. Sham-operated animals served as control ones. Electrical activity was recorded in the region of the Locus coeruleus, the dorsal hippocampus, and in the caudatus-putamen complex. Beginning from the second week after surgery in experimental rats was observed a generalization of the theta-rhythm (7-9 Hz) in electrical activity of the Locus coeruleus, hippocampus and the caudatus-putamen complex during orienting-exploratory activity. At the same time such a rhythm was enhanced in animals with motor and orienting-exploratory agitation. In comparison to the sham-operated animals there were no changes in expression in the electrical activity of the subcortical structures of the rhythm of 5-6 Hz, accompanying immobilization state as well as that of 9-11 Hz, accompanying teeth chattering. The dynamics of changes of subcortical electrical activity of rats after the Locus coeruleus destruction may serve an index of the functional state of the animals under the conditions of noradrenergic deficiency. Changes of the brain electrical activity and behaviour of the animals revealed are considered to be a consequence of a relative enhancement of the central serotoninergic regulation of the subcortical structures being caused by a weakening of noradrenergic influences.


Sleep deprivation induces corneal epithelial progenitor cell over-expansion through disruption of redox homeostasis in the tear film.
Sleep deficiency, a common public health problem, causes ocular discomfort and affects ocular surface health. However, the underlying mechanism remains unclear. Herein, we identified that short-term sleep deprivation (SD) resulted in hyperproliferation of corneal epithelial progenitor cells (CEPCs) in mice. The expression levels of p63 and Keratin 14, the biomarkers of CEPCs, were upregulated in the corneal epithelium after short-term SD. In addition, SD led to elevated levels of reactive oxygen species (ROS), and subsequent decrease in antioxidant capacity, in the tear film. Exogenous hydrogen peroxide (H2O2) could directly stimulate the proliferation of CEPCs in vivo and in vitro. Topical treatment of antioxidant L-glutathione preserved the over-proliferation of CEPCs and attenuated corneal epithelial defects in SD mice. Moreover, the activation of the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway is essential to ROS-stimulated cell proliferation in CEPCs. However, long-term SD ultimately led to early manifestation of limbal stem cell deficiency.


Social-cognitive and school factors in lifetime smoking among adolescents.
Smoking is a serious health threat and identifying risk factors for smoking is thus of great importance. The aim of the study was to examine the effects of social-cognitive factors and school factors on lifetime smoking status among adolescents. The study was based on cross-sectional data on 2,913 Danish adolescents in grade 7 attending 118 randomly selected public schools. Social-cognitive factors were examined with five measures: self-efficacy to resist pressure to smoke, social influence (norms), social influence (behavior), social influence (pressure), and attitude. We used multilevel analyses to estimate the associations between social-cognitive factors and lifetime smoking status as well as the group-level effects of school, school class, and gender group in the school class. Each social-cognitive factor was significantly associated with lifetime smoking status, even when several potential confounders and the effects of school, school class, and gender group were taken into account. Of the three group-level school factors, gender group in the school class had the strongest effect on smoking status. We conclude that self-efficacy to resist pressure to smoke, attitude, and the three types of social influence are significantly associated with lifetime smoking status, even when the effects of group-level school factors are taken into account. The strong effect of gender group in school class on lifetime smoking status indicates that prevention actions should address the social context of adolescents.


BOCS: DNA k-mer content and scoring for rapid genetic biomarker identification at low coverage.
A single, inexpensive diagnostic test capable of rapidly identifying a wide range of genetic biomarkers would prove invaluable in precision medicine. Previous work has demonstrated the potential for high-throughput, label-free detection of A-G-C-T content in DNA k-mers, providing an alternative to single-letter sequencing while also having inherent lossy data compression and massively parallel data acquisition. Here, we apply a new bioinformatics algorithm - block optical content scoring (BOCS) - capable of using the high-throughput content k-mers for rapid, broad-spectrum identification of genetic biomarkers. BOCS uses content-based sequence alignment for probabilistic mapping of k-mer contents to gene sequences within a biomarker database, resulting in a probability ranking of genes on a content score. Simulations of the BOCS algorithm reveal high accuracy for identification of single antibiotic resistance genes, even in the presence of significant sequencing errors (100% accuracy for no sequencing errors, and >90% accuracy for sequencing errors at 20%), and at well below full coverage of the genes. Simulations for detecting multiple resistance genes within a methicillin-resistant Staphylococcus aureus (MRSA) strain showed 100% accuracy at an average gene coverage of merely 0.515, when the k-mer lengths were variable and with 4% sequencing error within the k-mer blocks. Extension of BOCS to cancer and other genetic diseases met or exceeded the results for resistance genes. Combined with a high-throughput content-based sequencing technique, the BOCS algorithm potentiates a test capable of rapid diagnosis and profiling of genetic biomarkers ranging from antibiotic resistance to cancer and other genetic diseases.


Mycotic Aneurysm of the Middle Cerebral Artery Leading to Subarachnoid Hemorrhage, as the Initial Presentation of Bartonella henselae Endocarditis.
Bartonella species was first reported as a cause of endocarditis in 1993, currently it is thought to account for 3-4 percent of all diagnosed cases. Initial symptoms of Bartonella endocarditis are non-specific like weight loss, fever and fatigue. There are very few reported cases of Bartonella endocarditis causing mycotic aneurysm. We present a case of a 60-year-old male who presented with subarachnoid hemorrhage secondary to mycotic aneurysm. Due to high suspicion of endocarditis leading to mycotic aneurysm he underwent transesophageal echocardiography which showed mitral valve vegetations. His blood cultures were negative, he was eventually diagnosed with Bartonella henselae by elevated IgG titers greater than 1:800. Due to repeated mycotic aneurysms on antibiotics, he underwent surgical mitral valve replacement along with the full course of antibiotics and has been asymptomatic since.


Inhibition by local anaesthetics of adenine nucleotide translocation in rat liver mitochondria.
1. The mechanism of adenine nucleotide translocation in mitochondria isolated from rat liver was further examined by using the local anaesthetics procaine, butacaine, nupercaine and tetracaine as perturbators of lipid-protein interactions. Each of these compounds inhibited translocation of ADP and of ATP; butacaine was the most effective with 50% inhibition occurring at 30mum for 200mum-ATP and at 10mum for 200mum-ADP. The degree of inhibition by butacaine of both adenine nucleotides was dependent on the concentration of adenine nucleotide present; with low concentrations of adenine nucleotide, low concentrations of butacaine-stimulated translocation, but at high concentrations (greater than 50mum) low concentrations of butacaine inhibited translocation. Butacaine increased the affinity of the translocase for ATP to a value which approached that of ADP. 2. Higher concentrations of nupercaine and of tetracaine were required to inhibit translocation of both nucleotides; 50% inhibition of ATP translocation occurred at concentrations of 0.5mm and 0.8mm of these compounds respectively. The pattern of inhibition of ADP translocation by nupercaine and tetracaine was more complex than that of ATP; at very low concentrations (less than 250mum) inhibition ensued, followed by a return to almost original rates at 1mm. At higher concentrations inhibition of ADP translocation resulted. 3. That portion of ATP translocation stimulated by Ca(2+) was preferentially inhibited by each of the local anaesthetics tested. In contrast, inhibition by the anaesthetics of ADP translocation was prevented by low concentrations of Ca(2+). 4. The data provide further support for our hypothesis that lipid-protein interactions are important determinants in the activity of the adenine nucleotide translocase in mitochondria.


Thin film biosensor for rapid visual detection of nucleic acid targets.
We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets. In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector oligonucleotide allows initiation of formation of an organic thin film on the biosensor. Thin film formation is completed by enzymatic activity of peroxidase conjugated to an anti-biotin antibody. Peroxidase catalyzes deposition of an insoluble product onto the silicon surface, generating a uniform thin film. The increased thickness on the surface alters the perceived color of the biosensor through changes in the interference patterns of reflected light from the surface, causing a color change from gold to purple. The biosensor results may be evaluated by direct visual inspection or quantified by ellipsometry. Results are obtained in 25 min with a detection limit of 5 pmol/L (150 amol/sample). Selectivity of the biosensor is demonstrated by discrimination of single nucleotide mismatches. Multitarget arrays are also analyzed with the thin film biosensor, and the system is capable of detecting targets from human serum and urine. The biosensor surface is inexpensive to produce, and the assay format is simple and rapid. The thin film biosensor is adaptable to a wide variety of nucleic acid detection applications, including rapid diagnostic testing for infectious disease panels, antibiotic resistance panels, or allelic discrimination of specific genetic markers.


Structural Optimization of Antimycobacterial Azaaurones Towards Improved Solubility and Metabolic Stability.
While N-acetyl azaaurones have already been disclosed for their potential against tuberculosis (TB), their low metabolic stability remains an unaddressed liability. We now report a study designed to improve the metabolic stability and solubility of the azaaurone scaffold and to identify the structural requirements for antimycobacterial activity. Replacing the N-acetyl moiety for a N-carbamoyl group led to analogues with sub- and nanomolar potencies against M. tuberculosis H37Rv, as well as equipotent against drug-susceptible and drug-resistant M. tuberculosis isolates. The new N-carbamoyl azaaurones exhibited improved microsomal stability, compared to their N-acetylated counterparts, with several compounds displaying moderate to high kinetic solubility. The frequency of spontaneous resistance to azaaurones was observed to be in the range of 10-8 , a value that is comparable to current TB drugs in the market. Overall, these results reveal that azaaurones are amenable to structural modifications to improve metabolic and solubility liabilities, and highlight their potential as antimycobacterial agents.


Formation of bimetallic Ag-Au nanowires by metallization of artificial DNA duplexes.
Uniform bimetallic nanowires, tunable in size, have been grown on artificial DNA templates via a two-step metallization process. Alkyne-modified cytosines were incorporated into 900-base-pair polymerase-chain-reaction fragments. The alkyne modifications serve as addressable metal-binding sites after conversion to a sugar triazole derivative via click chemistry. Reaction of the Tollens reagent with these sugar-coated DNA duplexes generates Ag0 metallization centers around the sugar modification sites of the DNA. After a subsequent enhancement step using gold, nanowires < or = 10 nm in diameter with a homogeneous surface profile were obtained. Furthermore, the advantage of this two-step procedure lies in the high selectivity of the process, due to the exact spatial control of modified DNA base incorporation and hence the confinement of metallization centers at addressable sites. Besides experiments on a membrane as a proof for the selectivity of the method, atomic force microscopy (AFM) studies of the wires produced on Si-SiO2 surfaces are discussed. Furthermore, we demonstrate time-dependent metallization experiments, monitored by AFM.


Object Boundary Detection in Natural Images May Depend on "Incitatory" Cell-Cell Interactions.
Detecting object boundaries is crucial for recognition, but how the process unfolds in visual cortex remains unknown. To study the problem faced by a hypothetical boundary cell, and to predict how cortical circuitry could produce a boundary cell from a population of conventional "simple cells," we labeled 30,000 natural image patches and used Bayes' rule to help determine how a simple cell should influence a nearby boundary cell depending on its relative offset in receptive field position and orientation. We identified the following three basic types of cell-cell interactions: rising and falling interactions with a range of slopes and saturation rates, and nonmonotonic (bump-shaped) interactions with varying modes and amplitudes. Using simple models, we show that a ubiquitous cortical circuit motif consisting of direct excitation and indirect inhibition-a compound effect we call "incitation"-can produce the entire spectrum of simple cell-boundary cell interactions found in our dataset. Moreover, we show that the synaptic weights that parameterize an incitation circuit can be learned by a single-layer "delta" rule. We conclude that incitatory interconnections are a generally useful computing mechanism that the cortex may exploit to help solve difficult natural classification problems.SIGNIFICANCE STATEMENT Simple cells in primary visual cortex (V1) respond to oriented edges and have long been supposed to detect object boundaries, yet the prevailing model of a simple cell-a divisively normalized linear filter-is a surprisingly poor natural boundary detector. To understand why, we analyzed image statistics on and off object boundaries, allowing us to characterize the neural-style computations needed to perform well at this difficult natural classification task. We show that a simple circuit motif known to exist in V1 is capable of extracting high-quality boundary probability signals from local populations of simple cells. Our findings suggest a new, more general way of conceptualizing cell-cell interconnections in the cortex.


Solid-phase microextraction using poly(pyrrole) film and liquid chromatography with UV detection for analysis of antidepressants in plasma samples.
Poly(pyrrole) (PPY) coating was prepared on a stainless-steel (SS) wire for solid-phase microextraction (SPME) by electrochemical deposition (cyclic voltammetric). The PPY was evaluated by analyzing new-generation antidepressants (mirtazapine, citalopram, paroxetine, duloxetine, fluoxetine, and sertraline) in plasma sample by SPME and liquid chromatography with UV detection (LC-UV). The effect of electrolyte solution (lithium perchlorate or tetrabutylammonium perchlorate) and the number of cycles (50, 100 or 200) applied during the polymerization process on the SPME performance was evaluated. Important factors in the optimization of SPME efficiency such as extraction time, temperature, pH, influence of plasma proteins on sorption mechanisms, and desorption conditions are discussed. The SPME-PPY/LC method showed to be linear in concentrations ranging from the limit of quantification (LOQ) to 1200 ng mL(-1). The LOQ values range from 16 to 25 ng mL(-1). The inter-day precision of the SPME-PPY/LC method presented coefficient of variation (CV) lower than 15%. Based on analytical validation results, the SPME-PPY/LC methodology showed to be adequate for antidepressant analysis, from therapeutic to toxic levels. In order to evaluate the proposed method for clinical use, the SPME-PPY/LC method was applied to the analysis of plasma samples from elderly depressed patients.


Modeling the future of irrigation: A parametric description of pressure compensating drip irrigation emitter performance.
Drip irrigation is a means of distributing the exact amount of water a plant needs by dripping water directly onto the root zone. It can produce up to 90% more crops than rain-fed irrigation, and reduce water consumption by 70% compared to conventional flood irrigation. Drip irrigation may enable millions of poor farmers to rise out of poverty by growing more and higher value crops, while not contributing to overconsumption of water. Achieving this impact will require broadening the engineering knowledge required to design new, low-cost, low-power drip irrigation technology, particularly for poor, off-grid communities in developing countries. For more than 50 years, pressure compensating (PC) drip emitters-which can maintain a constant flow rate under variations in pressure, to ensure uniform water distribution on a field-have been designed and optimized empirically. This study presents a parametric model that describes the fluid and solid mechanics that govern the behavior of a common PC emitter architecture, which uses a flexible diaphragm to limit flow. The model was validated by testing nine prototypes with geometric variations, all of which matched predicted performance to within R2 = 0.85. This parametric model will enable irrigation engineers to design new drip emitters with attributes that improve performance and lower cost, which will promote the use of drip irrigation throughout the world.


Lactate stimulates vasculogenic stem cells via the thioredoxin system and engages an autocrine activation loop involving hypoxia-inducible factor 1.
The recruitment and differentiation of circulating stem/progenitor cells (SPCs) in subcutaneous Matrigel in mice was assessed. There were over one million CD34(+) SPCs per Matrigel plug 18 h after Matrigel implantation, and including a polymer to elevate the lactate concentration increased the number of SPCs by 3.6-fold. Intricate CD34(+) cell-lined channels were linked to the systemic circulation, and lactate accelerated cell differentiation as evaluated based on surface marker expression and cell cycle entry. CD34(+) SPCs from lactate-supplemented Matrigel exhibited significantly higher concentrations of thioredoxin 1 (Trx1) and hypoxia-inducible factor 1 (HIF-1) than cells from unsupplemented Matrigel, whereas Trx1 and HIF-1 in CD45(+) leukocytes were not elevated by lactate. Results obtained using small inhibitory RNA (siRNA) specific to HIF-1 and mice with conditionally HIF-1 null myeloid cells indicated that SPC recruitment and lactate-mediated effects were dependent on HIF-1. Cells from lactate-supplemented Matrigel had higher concentrations of phosphorylated extracellular signal-regulated kinases 1 and 2, Trx1, Trx reductase (TrxR), vascular endothelial growth factor (VEGF), and stromal cell-derived factor 1 (SDF-1) than cells from unsupplemented Matrigel. SPC recruitment and protein changes were inhibited by siRNA specific to lactate dehydrogenase, TrxR, or HIF-1 and by oxamate, apocynin, U0126, N-acetylcysteine, dithioerythritol, and antibodies to VEGF or SDF-1. Oxidative stress from lactate metabolism by SPCs accelerated further SPC recruitment and differentiation through Trx1-mediated elevations in HIF-1 levels and the subsequent synthesis of HIF-1-dependent growth factors.


Risk Stratification for Acute Kidney Injury: Are Biomarkers Enough?
Acute kidney injury (AKI) is a common and serious complication that is associated with several adverse outcomes in hospitalized patients. AKI significantly increases the risk of mortality, need for renal replacement therapy, and intensive care admission, and it also has serious economic ramifications. Effective risk stratification to identify patients at risk for severe AKI is essential for targeting our health care and research resources to tackle this important public health issue. The overwhelming majority of research in earlier diagnosis and risk stratification of AKI over the past 10 years has focused on novel biomarker development. The purpose of this review is to provide an update on other novel risk stratification tools than can be used in the prognostication of AKI. We discuss the utility of the furosemide stress test in predicting the severity of AKI and the renal angina index in predicting the occurrence of AKI. We also discuss NephroCheck, a prognostic test that measures tissue inhibitor of metalloproteinase-2 and insulin-like growth factor binding protein 7 for the early detection of severe AKI.


The proteinase activated receptor-2 (PAR-2) mediates mitogenic responses in human vascular endothelial cells.
Proteolytically cleaved receptors, typified by the functional thrombin receptor (TR), represent a novel class of receptors that mediate signaling events by functional coupling to G proteins. Northern blot analysis completed with a human proteinase activated receptor-2 (PAR-2) cDNA as probe demonstrated the approximately 3.5kb PAR-2 transcript in total cellular RNA from human umbilical vein endothelial cells (HUVEC). Microspectrofluorimetry using Fura2-loaded HUVEC demonstrated a dose-dependent elevation in intracellular calcium transients ([Ca2+]i) to murine PAR39-44 (SLIGRL, putative neoligand after cleavage), with an approximate EC50 of 30 microM, and evidence for homologous desensitization with complete recovery at 45 min. Xenopus oocytes microinjected with TR cRNA failed to respond to 200 microM PAR39-44, and TR-targeted antisense oligonucleotides specifically abrogated thrombin-induced but not PAR39-44-mediated [Ca2+]i, excluding the possibility that TR/PAR-2 cell-surface coexpression was structurally linked. HUVEC incubated with PAR39-44 demonstrated a dose- and time-dependent mitogenic response similar to that seen with thrombin or TR42-47 (TR-activating peptide, SFLLRN). Preactivation of HUVEC with either PAR39-44 or thrombin resulted in heterologous desensitization to the corresponding agonist, an effect that was mediated primarily by TR internalization as evaluated by immunofluorescence and quantitative ELISA. These results ascribe a previously unrecognized function to the PAR-2 receptor, imply that a natural enzyme agonist may circulate in plasma, and suggest the presence of an additional regulatory mechanism controlling receptor activation events in vascular endothelial cells.


Resistin promotes cardiac hypertrophy via the AMP-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) and c-Jun N-terminal kinase/insulin receptor substrate 1 (JNK/IRS1) pathways.
Resistin has been suggested to be involved in the development of diabetes and insulin resistance. We recently reported that resistin is expressed in diabetic hearts and promotes cardiac hypertrophy; however, the mechanisms underlying this process are currently unknown. Therefore, we wanted to elucidate the mechanisms associated with resistin-induced cardiac hypertrophy and myocardial insulin resistance. Overexpression of resistin using adenoviral vector in neonatal rat ventricular myocytes was associated with inhibition of AMP-activated protein kinase (AMPK) activity, activation of tuberous sclerosis complex 2/mammalian target of rapamycin (mTOR) pathway, and increased cell size, [(3)H]leucine incorporation (i.e. protein synthesis) and mRNA expression of the hypertrophic marker genes, atrial natriuretic factor, brain natriuretic peptide, and β-myosin heavy chain. Activation of AMPK with 5-aminoimidazole-4-carbozamide-1-β-D-ribifuranoside or inhibition of mTOR with rapamycin or mTOR siRNA attenuated these resistin-induced changes. Furthermore, resistin increased serine phosphorylation of insulin receptor substrate (IRS1) through the activation of the apoptosis signal-regulating kinase 1/c-Jun N-terminal Kinase (JNK) pathway, a module known to stimulate insulin resistance. Inhibition of JNK (with JNK inhibitor SP600125 or using dominant-negative JNK) reduced serine 307 phosphorylation of IRS1. Resistin also stimulated the activation of p70(S6K), a downstream kinase target of mTOR, and increased phosphorylation of the IRS1 serine 636/639 residues, whereas treatment with rapamycin reduced the phosphorylation of these residues. Interestingly, these in vitro signaling pathways were also operative in vivo in ventricular tissues from adult rat hearts overexpressing resistin. These data demonstrate that resistin induces cardiac hypertrophy and myocardial insulin resistance, possibly via the AMPK/mTOR/p70(S6K) and apoptosis signal-regulating kinase 1/JNK/IRS1 pathways.