A pharmacovigilance study of antihypertensive medicines at a South delhi hospital.
The aim of the present study was to monitor adverse drug reactions associated with antihypertensive drugs. The study was conducted in medicine out patient department of 150-bed Majeedia Hospital at Hamdard University Campus in New Delhi. The study was conducted by way of one to one patient interview by a registered pharmacist using a questionnaire-based Adverse Drug Reaction Monitoring Form drafted according to the World Health Organisation Monitoring Guidelines. A total of 34 adverse drug reactions were observed in 250 hypertensive patients during the four month study. A high percentage of adverse drug reactions occurred in middle aged and female patients. Of the 34 adverse drug reactions, 18 (52.9%) were mild, 14 (41.2%) moderate and only 2 (5.8%) were classified as severe. Combination therapy was associated with significantly high occurrence (P < 0.05) of adverse drug reactions, with a total of 21 (61.8%) as compared to monotherapy (n=13, 38.2%). Cardiovascular adverse drug reactions constituted a major component, followed by gastrointestinal and respiratory complaints. Beta-blockers were the drug category associated with majority of adverse drug reactions, followed by angiotensin-converting enzyme inhibitors and calcium channel blockers. The above pharmacovigilance study presents the adverse drug reaction profile of antihypertensive medicines prescribed in our University Teaching Hospital. It was concluded that calcium channel blockers were the most frequently prescribed drug category but beta blockers were associated with higher frequency of adverse drug reactions.


Development of a Novel Simplified PBPK Absorption Model to Explain the Higher Relative Bioavailability of the OROS® Formulation of Oxybutynin.
A new minimal Segmented Transit and Absorption model (mSAT) model has been recently proposed and combined with intrinsic intestinal effective permeability (P eff,int ) to predict the regional gastrointestinal (GI) absorption (f abs ) of several drugs. Herein, this model was extended and applied for the prediction of oral bioavailability and pharmacokinetics of oxybutynin and its enantiomers to provide a mechanistic explanation of the higher relative bioavailability observed for oxybutynin's modified-release OROS® formulation compared to its immediate-release (IR) counterpart. The expansion of the model involved the incorporation of mechanistic equations for the prediction of release, transit, dissolution, permeation and first-pass metabolism. The predicted pharmacokinetics of oxybutynin enantiomers after oral administration for both the IR and OROS® formulations were in close agreement with the observed data. The predicted absolute bioavailability for the IR formulation was within 5% of the observed value, and the model adequately predicted the higher relative bioavailability observed for the OROS® formulation vs. the IR counterpart. From the model predictions, it can be noticed that the higher bioavailability observed for the OROS® formulation was mainly attributable to differences in the intestinal availability (F G ) rather than due to a higher colonic f abs , thus confirming previous hypotheses. The predicted f abs was almost 70% lower for the OROS® formulation compared to the IR formulation, whereas the F G was almost eightfold higher than in the IR formulation. These results provide further support to the hypothesis of an increased F G as the main factor responsible for the higher bioavailability of oxybutynin's OROS® formulation vs. the IR.


CTLA-4 blockade enhances the CTL responses to the p53 self-tumor antigen.
p53 is an attractive target for cancer immunotherapy because it is overexpressed in a high proportion of many different types of tumors. However, it is also expressed in normal tissues and acts as a toleragen in vivo. Previously, detailed examination of the repertoire specific for the murine p53(261-269) epitope in conventional and p53-deficient mice demonstrated that because of expression of p53, the CD8(+) T cells that respond to this epitope express low-affinity TCRs. It has been reported that tolerance to tumor Ags can be broken by in vivo administration of anti-CTLA-4 mAb. With the goal of overriding tolerance and achieving optimal activation of p53-specific CTL, the current study has assessed the effect of anti-CTLA-4 mAb on the p53-specific repertoire. It was found that blockade of CTLA-4 engagement at the time of antigenic stimulation induced a vigorous amplification of the CTL responses to p53 as well as proportionate expansion of the memory T cell pool. This effect was dependent on the presence of CD4(+) T cell help and correlated with an enhancement of helper function. However, anti-CTLA-4 treatment did not enhance the avidity of the resultant p53-specific CTL populations and, therefore, could not reverse this important consequence of tolerance.


Anti-RAGE antibody attenuates isoflurane-induced cognitive dysfunction in aged rats.
Several animal studies demonstrated that the volatile anesthetic isoflurane could influence the blood-brain barrier (BBB) integrity, which involved the cognitive impairment. Increasing evidence has also shown that the receptor for advanced glycation end-products (RAGE) played a major role in maintaining the integrity of BBB. The present study aimed to determine whether the RAGE-specific antibody protects against BBB disruption and cognitive impairment induced by isoflurane exposure in aged rats. 108 aged rats were randomly divided into four groups: (1) control group (Control); (2) 4h of 2% isoflurane exposure group (ISO); (3) RAGE antibody (20μL, 2.5μg/μL) treated+4h of 2% isoflurane exposure group (anti-RAGE+ISO); (4) RAGE antibody (20μL, 2.5μg/μL) treated group (anti-RAGE). The isoflurane anesthesia resulted in the upregulation of hippocampal RAGE expression, disruption of BBB integrity, neuroinflammation, and beta-amyloid (Aβ) accumulation in aged rats. In addition, significant cognitive deficits in the Morris water maze test was also observed. The antibody pretreatment resulted in significant improvements in BBB integrity. Furthermore, the expression of RAGE and proinflammatory mediators, as well as, Aβ accumulation were attenuated. Moreover, the antibody administration attenuated the isoflurane-induced cognitive impairment in aged rats. These results demonstrate that RAGE signaling is involved in BBB damage after isoflurane exposure. Thus, the RAGE antibody represents a novel therapeutic intervention to prevent isoflurane-induced cognitive impairment.


[Immunopharmacological activity of xymedone in patients with atopic bronchial asthma].
Xymedone produced therapeutic effect in patients with atopic bronchial asthma. The drug action was manifested by a decrease in the IgE level and an increase in gamma-interferon level. Adding xymedone to lymphocytes cultivated with phytohemagglutinin revealed stimulating action of this drug on the delayed-type hypersensitivity. Mechanisms involved in the immunotropic action of xymedone are discussed.


Genetic analyses of Schizosaccharomyces pombe dna2(+) reveal that dna2 plays an essential role in Okazaki fragment metabolism.
In this report, we investigated the phenotypes caused by temperature-sensitive (ts) mutant alleles of dna2(+) of Schizosaccharomyces pombe, a homologue of DNA2 of budding yeast, in an attempt to further define its function in vivo with respect to lagging-strand synthesis during the S-phase of the cell cycle. At the restrictive temperature, dna2 (ts) cells arrested at late S-phase but were unaffected in bulk DNA synthesis. Moreover, they exhibited aberrant mitosis when combined with checkpoint mutations, in keeping with a role for Dna2 in Okazaki fragment maturation. Similarly, spores in which dna2(+) was disrupted duplicated their DNA content during germination and also arrested at late S-phase. Inactivation of dna2(+) led to chromosome fragmentation strikingly similar to that seen when cdc17(+), the DNA ligase I gene, is inactivated. The temperature-dependent lethality of dna2 (ts) mutants was suppressed by overexpression of genes encoding subunits of polymerase delta (cdc1(+) and cdc27(+)), DNA ligase I (cdc17(+)), and Fen-1 (rad2(+)). Each of these gene products plays a role in the elongation or maturation of Okazaki fragments. Moreover, they all interacted with S. pombe Dna2 in a yeast two-hybrid assay, albeit to different extents. On the basis of these results, we conclude that dna2(+) plays a direct role in the Okazaki fragment elongation and maturation. We propose that dna2(+) acts as a central protein to form a complex with other proteins required to coordinate the multienzyme process for Okazaki fragment elongation and maturation.


Decreased bone mineral density in patients with pustulosis palmaris et plantaris.
Focal skeletal changes in patients with pustulosis palmaris et plantaris (PPP) are known to occur in a minority of patients. It is unknown whether these changes are unique events or whether they merely represent more progressed cases. The study was undertaken in order to investigate possible diffuse bone changes in patients with PPP. Bone mineral density (BMD) and biochemical markers of bone turnover were studied in 18 female patients and 18 age-matched controls. A significant correlation was found between lower-forearm BMD and disease duration (< 0.05). With increasing age, patients had significantly lower values than controls for both forearm and spine BMD (p < 0.005); patients who had PPP for more than 2 years had significantly reduced forearm BMD (19.8%; 95% CI: 5.6-32.8%) and spine BMD (17.4%; 95% CI: 0.9-33.8%). No significant differences were observed in biochemical markers of bone turnover, physical activity or body mass index between patients and controls. The proportion of smokers among patients was four times higher than among controls (p < 0.0005). No significant dose effect was found between number of pack-years and BMD. Although we cannot exclude that prolonged use of topical steroids under occlusion is a confounding factor, the study suggests that PPP patients have decreased BMD due to primary pathogenic events, and that osteoporosis may be an additional problem for these patients.


Utility of the frontal plane projection angle in females with patellofemoral pain.
Case-control study of females with patellofemoral pain syndrome (PFPS) and a control group. Three different approaches were used to examine the utility of a 2-dimensional (2-D) frontal plane projection angle (FPPA) measure of knee alignment. First, we measured the FPPA association with respect to 3-dimensional (3-D) lower extremity joint rotations during single-leg squats. Second, we determined the correlation of the FPPA during single-leg squats with hip and knee joint rotations during running and single leg jumping. Third, we compared the FPPA between females with and without PFPS. PFPS is associated with altered lower extremity kinematics during weight-bearing activities that decrease retropatellar contact area and increase retropatellar stress. An objective and simple procedure to quantify altered kinematics during weight-bearing activities may help clinicians identify individuals who may likely benefit from interventions to improve lower extremity kinematics. Twenty females with PFPS and 20 healthy female controls performed single-leg squats, running, and repetitive single-leg jumps while 3-D lower extremity kinematics were recorded. The FPPA was recorded by a digital camera during single-leg stance and single-leg squats. Correlation coefficients were used to quantify the association between the FPPA and transverse and frontal plane hip and knee angles for all activities. Independent t tests were used to compare FPPA values between groups. FPPA values representing medial displacement of the knee during single-leg squats were associated with increased hip adduction (r = 0.32 to 0.38, P<.044) and knee external rotation (r = 0.48 to 0.55, P<.001) across activities. FPPA values for the PFPS group reveal greater medial displacement of the knee compared with those of the control group during single-leg squats (P = .012). The association between the FPPA and lower extremity kinematics that are associated with PFPS suggest that the FPPA during single-leg squats may be a useful clinical measure. However, these methods should not be used to quantify 3-D joint rotations.


Patient perspectives of lithium and quetiapine augmentation treatment in treatment-resistant depression: A qualitative assessment.
Treatment-resistant depression (TRD) has a profound cost to patients and healthcare services worldwide. Pharmacological augmentation is one therapeutic option for TRD, with lithium and quetiapine currently recommended as first-line agents. Patient opinions about pharmacological augmentation may affect treatment outcomes, yet these have not been systematically explored. This study aimed to qualitatively assess patient experiences of lithium and quetiapine augmentation. Semi-structured interviews were conducted with 32 patients from the ongoing lithium versus quetiapine open-label trial comparing these augmentation agents in patients with TRD. Interviews were audio recorded, transcribed and a thematic analysis was used to assess patient opinions of each agent. Four main themes were generated from the thematic analysis: 'Initial concerns', 'Experience of side effects', 'Perception of treatment efficacy' and 'Positive perception of treatment monitoring'. Patient accounts indicated a predominantly positive experience of lithium and quetiapine augmentation. Greater apprehension about side effects was reported for lithium prior to treatment initiation, but greater experience of negative side effects was reported for quetiapine. Clinical monitoring was perceived positively. Patient accounts suggested treatment augmentation with lithium or quetiapine was acceptable and helpful for most patients. However, anticipation and experiences of adverse side effects may prevent some patients from benefitting from these treatments.


Treatment failure and hospital readmissions in severe COPD exacerbations treated with azithromycin versus placebo - a post-hoc analysis of the BACE randomized controlled trial.
In the BACE trial, a 3-month (3 m) intervention with azithromycin, initiated at the onset of an infectious COPD exacerbation requiring hospitalization, decreased the rate of a first treatment failure (TF); the composite of treatment intensification (TI), step-up in hospital care (SH) and mortality. (1) To investigate the intervention's effect on recurrent events, and (2) to identify clinical subgroups most likely to benefit, determined from the incidence rate of TF and hospital readmissions. Enrolment criteria included the diagnosis of COPD, a smoking history of ≥10 pack-years and ≥ 1 exacerbation in the previous year. Rate ratio (RR) calculations, subgroup analyses and modelling of continuous variables using splines were based on a Poisson regression model, adjusted for exposure time. Azithromycin significantly reduced TF by 24% within 3 m (RR = 0.76, 95%CI:0.59;0.97, p = 0.031) through a 50% reduction in SH (RR = 0.50, 95%CI:0.30;0.81, p = 0.006), which comprised of a 53% reduction in hospital readmissions (RR = 0.47, 95%CI:0.27;0.80; p = 0.007). A significant interaction between the intervention, CRP and blood eosinophil count at hospital admission was found, with azithromycin significantly reducing hospital readmissions in patients with high CRP (> 50 mg/L, RR = 0.18, 95%CI:0.05;0.60, p = 0.005), or low blood eosinophil count (<300cells/μL, RR = 0.33, 95%CI:0.17;0.64, p = 0.001). No differences were observed in treatment response by age, FEV1, CRP or blood eosinophil count in continuous analyses. This post-hoc analysis of the BACE trial shows that azithromycin initiated at the onset of an infectious COPD exacerbation requiring hospitalization reduces the incidence rate of TF within 3 m by preventing hospital readmissions. In patients with high CRP or low blood eosinophil count at admission this treatment effect was more pronounced, suggesting a potential role for these biomarkers in guiding azithromycin therapy. ClinicalTrials.gov number. NCT02135354 .


Tamoxifen resistance in breast tumors is driven by growth factor receptor signaling with repression of classic estrogen receptor genomic function.
Not all breast cancers respond to tamoxifen, and many develop resistance despite initial benefit. We used an in vivo model of estrogen receptor (ER)-positive breast cancer (MCF-7 xenografts) to investigate mechanisms of this resistance and develop strategies to circumvent it. Epidermal growth factor receptor (EGFR) and HER2, which were barely detected in control estrogen-treated tumors, increased slightly with tamoxifen and were markedly increased when tumors became resistant. Gefitinib, which inhibits EGFR/HER2, improved the antitumor effect of tamoxifen and delayed acquired resistance, but had no effect on estrogen-stimulated growth. Phosphorylated levels of p42/44 and p38 mitogen-activated protein kinases (both downstream of EGFR/HER2) were increased in the tamoxifen-resistant tumors and were suppressed by gefitinib. There was no apparent increase in phosphorylated AKT (also downstream of EGFR/HER2) in resistant tumors, but it was nonetheless suppressed by gefitinib. Phosphorylated insulin-like growth factor-IR (IGF-IR), which can interact with both EGFR and membrane ER, was elevated in the tamoxifen-resistant tumors compared with the sensitive group. However, ER-regulated gene products, including total IGF-IR itself and progesterone receptor, remained suppressed even at the time of acquired resistance. Tamoxifen's antagonism of classic ER genomic function was retained in these resistant tumors and even in tumors that overexpress HER2 (MCF-7 HER2/18) and are de novo tamoxifen-resistant. In conclusion, EGFR/HER2 may mediate tamoxifen resistance in ER-positive breast cancer despite continued suppression of ER genomic function by tamoxifen. IGF-IR expression remains dependent on ER but is activated in the tamoxifen-resistant tumors. This study provides a rationale to combine HER inhibitors with tamoxifen in clinical studies, even in tumors that do not initially overexpress EGFR/HER2.


Roles and Mechanistic Bases of Glucocorticoid Regulation of Avian Reproduction.
To maximize fitness, organisms must invest energetic and nutritional resources into developing, activating, and maintaining reproductive physiology and behavior. Corticosterone (CORT), the primary avian glucocorticoid, regulates energetic reserves to meet metabolic demands. At low (baseline) plasma levels, CORT activates avian mineralocorticoid receptors and may stimulate lipid mobilization, foraging activity, and feeding behavior. During stress in birds, elevated plasma CORT also stimulates glucocorticoid receptors and may promote glycemia, lipolysis, and proteolysis. Furthermore, CORT orchestrates physiological and behavioral adjustments to perceived threats. While many avian studies demonstrate effects of CORT on reproduction, few studies have elucidated the mechanisms, including receptor activation and site(s) of action, which underlie these effects. Even fewer studies have investigated how low and elevated plasma CORT regulates energetic reserves to meet the metabolic demands of reproduction. Here, we propose several hypotheses to clarify the direct and indirect effects of CORT on avian reproductive physiology and behavior. In addition, we emphasize the need for new manipulative studies involving alterations of endogenous plasma CORT levels and/or food availability to elucidate how CORT regulates the energetic demands of reproduction.


A Dual Role of Osteopontin in Modifying B Cell Responses.
The occurrence of B cell aggregates within the central nervous system (CNS) has prompted the investigation of the potential sources of pathogenic B cell and T cell responses in a subgroup of secondary progressive multiple sclerosis (MS) patients. Nevertheless, the expression profile of molecules associated with these aggregates and their role in aggregate development and persistence is poorly described. Here, we focused on the expression pattern of osteopontin (OPN), which is a well-described cytokine, in MS brain tissue. Autopsied brain sections from MS cases with and without B cell pathology were screened for the presence of CD20+ B cell aggregates and co-expression of OPN. To demonstrate the effect of OPN on B cells, flow cytometry, ELISA and in vitro aggregation assays were conducted using the peripheral blood of healthy volunteers. Although OPN was expressed in MS brain tissue independent of B cell pathology, it was also highly expressed within B cell aggregates. In vitro studies demonstrated that OPN downregulated the co-stimulatory molecules CD80 and CD86 on B cells. OPN-treated B cells produced significantly lower amounts of IL-6. However, OPN-treated B cells also exhibited a higher tendency to form homotypic cell aggregates in vitro. Taken together, our data indicate a conflicting role of OPN in modulating B cell responses.


Antiphospholipid syndrome and pre-eclampsia.
Antiphospholipid syndrome (APS) is defined as an autoimmune disorder characterized by recurrent thrombosis or obstetrical morbidity. These features are linked to the presence in blood of autoantibodies against negatively charged phospholipids or phospholipid-binding proteins. Obstetric morbidity includes recurrent abortion (early and late) and severe pre-eclampsia (P-EC)/hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome, and/or severe placental insufficiency. Criteria that define the major clinical and laboratory events were published in revised forms in the Sydney recommendations in 2006. We analyzed the blood of patients with severe P-EC according to the subgroups based on the 2006 revised criteria definition and compared these results with women after uncomplicated pregnancy and delivery. We found 20% elevated antiphospholipid antibodies (APAs) in women with severe P-EC (group I, 7.5%; group IIa, 5.0%; group IIb, 5.0%; group IIc, 2.5%). The increased APAs were observed only in women with severe P-EC (odds ratio: 2.45; 95% confidence interval, 1.01 to 4.3) and not in patients with severe P-EC at >34 weeks of gestation. According to our retrospective observation, we recommend the determination of anticardiolipin antibodies, lupus anticoagulant, and β-2 glycoprotein-1 antibodies in patients with severe P-EC at <34 weeks of gestation.


Optimal Positioning of Inertial Measurement Units in a Smart Shirt for Determining Respiratory Volume.
Tidal volume can be estimated using the surface motions of the upper body induced by respiration. However, the precision and instrumentation of such estimation must be improved to allow widespread application. In this study, respiration induced changes in parameters that can be recorded with inertial measurement units are examined to determine tidal volumes. Based on the data of an optical motion capture system, the optimal positions of inertial measurement units (IMU) in a smart shirt for sets of 4, 5 or 6 sensors were determined. The errors observed indicate the potential to determine tidal volumes using IMUs in a smart shirt.Clinical Relevance- The measurement of respiratory volumes via a low-cost and unobtrusive smart shirt would be a major advance in clinical diagnostics. In particular, conventional methods are expensive, and uncomfortable for conscious patients if measurement is desired over an extended period. A smart-shirt based on inertial sensors would allow a comfortable measurement and could be used in many clinical scenarios - from sleep apnoea monitoring to homecare and respiratory monitoring of comatose patients.


Effect of post-ruminal guanidinoacetic acid supplementation on creatine synthesis and plasma homocysteine concentrations in cattle.
Creatine stores high-energy phosphate bonds in muscle, which is critical for muscle activity. In animals, creatine is synthesized in the liver from guanidinoacetic acid (GAA) with methylation by S-adenosylmethionine. Because methyl groups are used for the conversion of GAA to creatine, methyl group deficiency may occur as a result of GAA supplementation. With this study, the metabolic responses of cattle to post-ruminal supplementation of GAA were evaluated with and without methionine (Met) supplementation as a source of methyl groups. Six ruminally cannulated Holstein heifers (520 kg) were used in a split-plot design with treatments arranged as a 2 × 5 factorial. The main plot treatments were 0 or 12 g/d of l-Met arranged in a completely randomized design; three heifers received each main plot treatment throughout the entire experiment. Subplot treatments were 0, 10, 20, 30, and 40 g/d of GAA, with GAA treatments provided in sequence from lowest to highest over five 6-d periods. Treatments were infused continuously to the abomasum. Heifers were limit-fed twice daily a diet consisting of (dry matter basis) 5.3 kg/d rolled corn, 3.6 kg/d alfalfa hay, and 50 g/d trace-mineralized salt. Plasma Met increased (P < 0.01) when Met was supplemented, but it was not affected by supplemental GAA. Supplementing GAA linearly increased plasma arginine (% of total amino acids) and plasma concentrations of GAA and creatinine (P < 0.001). Plasma creatine was increased at all levels of GAA except when 40 g/d of GAA was supplemented with no Met (GAA-quadratic × Met, P = 0.07). Plasma homocysteine was not affected by GAA supplementation when heifers received 12 g/d Met, but it was increased when 30 or 40 g/d of GAA was supplemented without Met (GAA-linear × Met, P = 0.003); increases were modest and did not suggest a dangerous hyperhomocysteinemia. Urinary concentrations of GAA and creatine were increased by all levels of GAA when 12 g/d Met was supplemented; increasing GAA supplementation up to 30 g/d without Met increased urinary GAA and creatine concentrations, but 40 g/d GAA did not affect urine concentrations of GAA and creatine when no Met was supplemented. Overall, post-ruminal GAA supplementation increased creatine supply to cattle. A methyl group deficiency, demonstrated by modest increases in plasma homocysteine, became apparent when 30 or 40 g/d of GAA was supplemented, but it was ameliorated by 12 g/d Met.


The National Cancer Institute's intervention trials.
The National Cancer Institute (NCI) has increased its emphasis in cancer prevention by undertaking a number of human intervention trials. Special emphasis is placed on chemoprevention, diet, and smoking and tobacco use. Some 24 clinical trials are ongoing in chemoprevention to assess the role of specific chemicals (natural and synthetic) in preventing, inhibiting, or reversing carcinogenesis. In diet and nutrition, several macronutrient trials are underway. The most significant of these is examining the relationship between dietary fat and breast cancer. In 1982, the NCI initiated a broad intervention research effort in smoking and tobacco use which has resulted in 46 prevention and clinical trials covering 25 states and more than 200 cities. More recently, a North American community-based intervention trial was begun to test cessation strategies for heavy smokers in 22 different sites. All of these intervention research efforts are testing strategies which later can be applied to large target population thereby supporting NCI's ambitious goal to reduce cancer mortality in this nation 50% by the year 2000.


Direct determination of serum glycyrrhetic acid by a monoclonal antibody-based inhibition ELISA using ibuprofen for releasing serum albumin-bound glycyrrhetic acid.
We found that ibuprofen (IBU) had a potential for releasing serum albumin-bound glycyrrhetic acid (GA). Based on this observation, IBU was used to pretreat samples for the determination of serum GA levels by an inhibition ELISA. This method, termed IBU method was evaluated by the recovery of GA from human serum albumin (HSA) or normal human serum (NHS) that contained the exogeneously added GA (37-1000 ng/ml). The mean recovery of GA from HSA and NHS samples treated with IBU were 104.7 and 105.2%, respectively, whereas those without IBU pretreatment were 2.8 and 10.7%, respectively. Comparison of IBU method and chloroform extraction method revealed that the GA content of serum samples pretreated by each method were almost the same. These results suggest that IBU method is useful as a serum processing procedure for the determination of serum GA levels by an inhibition ELISA.


Profile for amyloid-beta and tau expression in primary cortical cultures from 3xTg-AD mice.
Advances in transgenic technology as well as in the genetics of Alzheimer disease (AD) have allowed the establishment of animal models that reproduce amyloid-beta plaques and neurofibrillary tangles, the main pathological hallmarks of AD. Among these models, 3xTg-AD mice harboring PS1 (M146V), APP (Swe) and tau (P301L) human transgenes provided the model that most closely mimics human AD features. Although cortical cultures from 3xTg-AD mice have been shown to present disturbances in intracellular [Ca(2+)] homeostasis, the development of AD pathology in vitro has not been previously evaluated. In the current work, we determined the temporal profile for amyloid precursor protein, amyloid-beta and tau expression in primary cortical cultures from 3xTg-AD mice. Immunocytochemistry and Western blot analysis showed an increased expression of these proteins as well as several phosphorylated tau isoforms with time in culture. Alterations in calcium homeostasis and cholinergic and glutamatergic responses were also observed early in vitro. Thus, 3x-TgAD cortical neurons in vitro provide an exceptional tool to investigate pharmacological approaches as well as the cellular basis for AD and related diseases.


Elicitor- and A23187-induced expression of WCK-1, a gene encoding mitogen-activated protein kinase in wheat.
Wheat cultured cells were used to study the role of Ca2+ in regulating protein kinases during the induction of defense-related genes by fungal elicitor treatments. Manipulation of intracellular Ca2+ concentrations by treatment with calcium ionophore A23187 in the presence of high extracellular Ca2+ resulted in the induction of mRNA expression of WCK-1, a gene encoding mitogen-activated protein (MAP) kinase. The induction of WCK-1 mRNA by A23187 did not occur when extracellular Ca2+ was chelated by 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). The WCK-1 mRNA was also induced by Typhula ishikariensis-derived elicitors, suggesting a possible involvement of WCK-1 in the plant defense response against pathogens. BAPTA and a calcium channel blocker, La3+, inhibited the elicitor-induced expression of the WCK-1 mRNA. A recombinant fusion protein of WCK-1 (GST-WCK-1) autophosphorylated at the Tyr residue and exhibited an autophosphorylation-dependent protein kinase activity towards myelin basic protein. Alteration of Tyr-196 in the conserved 'TEY' motif in GST-WCK-1 to Phe by site-directed mutagenesis abolished the autophosphorylation. The GST-WCK-1 protein was activated by elicitor-treated wheat cell extracts but not by the control extract. These results suggest that fungal elicitors activate WCK-1, a specific MAP kinase in wheat. Furthermore, the results suggest a possible involvement of Ca2+ in enhancing the MAP kinase signaling cascade in plants by controlling the levels of the MAP kinase transcripts.


Trial-to-trial variability of spike response of V1 and saccadic response time.
Single neurons in the primary visual cortex (V1) show variability in spike activity in response to an identical visual stimulus. In the current study, we examined the behavioral significance of the variability in spike activity of V1 neurons for visually guided saccades. We recorded single-cell activity from V1 of monkeys trained to detect and make saccades toward visual targets of varying contrast and analyzed trial-to-trial covariation between the onset time or firing rate of neural response and saccadic response time (RT). Neural latency (NL, the time of the first spike of neural response) was correlated with RT, whereas firing rate (FR) was not. When FR was computed with respect to target onset ignoring NL, a "false" correlation between FR and RT emerged. Multiple regression and partial correlation analyses on NL and FR for predictability of RT variability, as well as a simulation with artificial Poisson spike trains, supported the conclusion that the correlation between FR with respect to target onset and RT was mediated by a correlation between NL and RT, emphasizing the role of trial-to-trial variability of NL for extracting RT-related signals. We attempted to examine laminar differences in RT-related activity. Neurons recorded in the superficial layers tended to show a higher sensitivity to stimulus contrast and a lower correlation with RT compared with those in the lower layers, suggesting a sensory-to-motor transformation within V1 that follows the order of known anatomical connections. These results demonstrate that the trial-to-trial variability of neural response in V1 propagates to the stage of saccade execution, resulting in trial-to-trial variability of RT of a visually guided saccade.


Preparation of a "functional library" of African green monkey DNA fragments which substitute for the processing/polyadenylation signal in the herpes simplex virus type 1 thymidine kinase gene.
Fragments of African green monkey (Cercopithecus aethiops) DNA (3.5 to 18.0 kilobases) were inserted downstream from the thymidine kinase (TK, tk) coding region in pTK206/SV010, a gene construct which lacks both copies of the hexanucleotide 5'-AATAAA-3' and contains a simian virus 40 origin of replication, allowing it to replicate in Cos-1 cells. No polyadenylated tk mRNA was detected in Cos-1 cells transfected by pTK206/SV010. The ability of simian DNA fragments to restore tk gene expression was examined by measuring the incorporation of [125I]iododeoxycytidine into DNA in Cos-1 cells transfected by pTK206/SV010 insertion derivatives. tk gene expression was restored by the insertion in 56 of the 67 plasmids analyzed, and the level of expression equaled or exceeded that obtained with the wild-type tk gene in 30 of these. In all plasmids examined that showed restoration of tk gene expression, polyadenylated tk mRNA of discrete size was detected. The sizes of these tk mRNAs were consistent with the existence of processing and polyadenylation signals within the inserted DNA fragments. The frequency with which inserted fragments restored tk gene expression suggests that the minimal signal for processing and polyadenylation is a hexanucleotide (AAUAAA or a similar sequence). LTK- cells were biochemically transformed to TK+ with representative insertion constructs. pTK206/SV010 transformed LTK- cells at a very low frequency; the frequency of transformation with insertion derivatives was 40 to 12,000 times higher.


Lumbar stenosis with spondylolisthesis: current concepts of surgical treatment.
As degenerative changes progress in the aging population, more patients will present with degenerative spondylolisthesis. The clinical picture is usually of a woman older than 50 years of age with a degenerative lumbar spondylolisthesis at L5 with symptoms of neurogenic claudication or radiculopathy. Imaging studies will reveal a central recess, a lateral recess or both, and/or neuroforaminal stenosis. When patients do not respond to nonoperative treatments, surgical intervention is indicated. Most studies have concluded that the addition of an arthrodesis improves the clinical outcome. A recent longterm study showed that a solid fusion and an adequate decompression is the procedure offering the best outcome. The current authors will review the pathogenesis, clinical picture, and treatment recommendations for degenerative lumbar spondylolisthesis.


Validation of the VBLaST peg transfer task: a first step toward an alternate training standard.
The FLS trainer lacks objective and automated assessments of laparoscopic performance and requires a large supply of relatively expensive consumables. Virtual reality simulation has a great potential as a training and assessment tool of laparoscopic skills and can overcome some limitations of the FLS trainer. This study was carried out to assess the value of our Virtual Basic Laparoscopic Surgical Trainer (VBLaST(©)) in the peg transfer task compared to the FLS trainer and its ability to differentiate performance between novice, intermediate, and expert groups. Thirty subjects were divided into three groups: novices (PGY1-2, n = 10), intermediates (PGY3-4, n = 10), and experts (PGY5, surgical fellows and attendings, n = 10). All subjects performed ten trials of the peg transfer task on each simulator. Assessment of laparoscopic performance was based on FLS scoring while a questionnaire was used for subjective evaluation. The performance scores in the two simulators were correlated, though subjects performed significantly better in the FLS trainer. Experts performed better than novices only on the FLS trainer while no significant differences were observed between the other groups. Moreover, a significant learning effect was found on both trainers, with a greater improvement of performance on the VBLaST(©). Finally, 82.6% of the subjects preferred the FLS over the VBLaST(©) for surgical training which could be attributed to the novelty of the VR technology and existing deficiencies of the user interface for the VBLaST(©). This study demonstrated that the VBLaST(©) reproduced faithfully some aspects of the FLS peg transfer task (such as color, size, and shape of the peg board, etc.) while other aspects require additional development. Future improvement of the user interface and haptic feedback will enhance the value of the system as an alternative to the FLS as the standard training tool for laparoscopic surgery skills.


IL-12 receptor deficiency revisited: IL-23-mediated signaling is also impaired in human genetic IL-12 receptor beta1 deficiency.
IFN-gamma and IL-12 are crucial cytokines for cell-mediated immunity against intracellular pathogens. We have previously shown that human IL-12Rbeta1-deficiency leads to impaired IL-12 responsiveness and unusual susceptibility to infections due to mycobacteria and salmonellae. IL-23 is a cytokine with functions that partially overlap with those of IL-12. IL-23 consists of IL-12p40 and a novel p19 protein, and binds to a receptor complex comprising IL-12Rbeta1 and IL-23R. Thus, IL-12Rbeta1-deficiency may impair both IL-12- and IL-23 signaling, and both may contribute to the immunological phenotypes. To examine whether IL-12Rbeta1 is essential for IL-23 signaling in human T cells, we have studied IL-23 responsiveness of four IL-12Rbeta1-deficient individuals. Whereas IL-23 promoted IFN-gamma production by CD4(+) and CD8(+) T cells in controls, IL-12Rbeta1-deficient T cells lacked IL-23-induced IFN-gamma secretion, but responded normally to IL-2, IL-4, IL-15 and IL-18. We also show that induction of IFN-gamma production by IL-23 depends upon TCR-ligation and is enhanced by CD28-costimulation. Furthermore, IL-23 cooperates with IL-12 and IL-18 in promoting IFN-gamma production in controls, but not in patients. We conclude that IL-12Rbeta1-deficiency impairs IL-12- and IL-23-dependent signaling in human T cells. The syndrome caused by IL-12Rbeta1-deficiency thus needs to be reinterpreted as resulting from defective IL-12-as well as IL-23-mediated immunity.


Fibrate-modulated expression of fibrinogen, plasminogen activator inhibitor-1 and apolipoprotein A-I in cultured cynomolgus monkey hepatocytes -- role of the peroxisome proliferator-activated receptor-alpha.
Fibrates are used to lower plasma triglycerides and cholesterol levels in hyperlipidemic patients. In addition, fibrates have been found to alter the plasma concentrations of fibrinogen, plasminogen activator inhibitor-1 (PAI-1) and apolipoprotein A-I (apo A-I). We have investigated the in vitro effects of fibrates on fibrinogen, PAI-1 and apo A-I synthesis and the underlying regulatory mechanisms in primary monkey hepatocytes. We show that fibrates time- and dose-dependently increase fibrinogen and apo A-I expression and decrease PAI-1 expression in cultured cynomolgus monkey hepatocytes, the effects demonstrating different potency for different fibrates. After three consecutive periods of 24 h the most effective fibrate. ciprofibrate (at 1 mmol/l), increased fibrinogen and apo A-I synthesis to 356% and 322% of control levels, respectively. Maximum inhibition of PAI-1 synthesis was about 50% of control levels and was reached by 1 mmol/l gemfibrozil or ciprofibrate after 48 h. A ligand for the retinoid-X-receptor (RXR), 9-cis retinoic acid, and specific activators of the peroxisome proliferator-activated receptor-alpha (PPARalpha), Wy14,643 and ETYA, influenced fibrinogen, PAI-1 and apo A-I expression in a similar fashion, suggesting a role for the PPARalpha/RXRalpha heterodimer in the regulation of these genes. When comparing the effects of the various compounds on PPARalpha transactivation activity as determined in a PPARalpha-sensitive reporter gene system and the ability of the compounds to affect fibrinogen, PAI-1 and apo A-I antigen production, a good correlation (r=0.80; p <0.01) between PPARalpha transactivation and fibrinogen expression was found. Apo A-I expression correlated only weakly with PPARalpha transactivation activity (r=0.47; p=0.24), whereas such a correlation was absent for PAI-1 (r=0.03; p=0.95). These results strongly suggest an involvement of PPARalpha in the regulation of fibrinogen gene expression.


A new endemic focus of Heterophyes nocens, Pygidiopsis summa, and other intestinal flukes in a coastal area of Muan-gun, Chollanam-do.
A small coastal village of Muan-gun, Chollanam-do, was surveyed for intestinal fluke infections, especially heterophyids such as Heterophyes nocens and Pygidiopsis summa by fecal examination on 108 inhabitants. The egg positive rate of heterophyids was very high, 75.0%, and that of other parasites was comparatively low, 0.9-3.7% by parasite species. After treatment of 20 patients showing high E.P.G. with praziquantel and purging with MgSO4, total 3,864 specimens of H. nocens were collected from the diarrheic stools of all the patients treated (3-1,338 individually) and total 703 P. summa were harvested from 18 patients (1-170 individually), together with several other species of flukes. Other flukes included Stictodora fuscata (164 specimens from 4 patients), Heterophyopsis continua (2 from 2 patients), and Gymnophalloides seoi (4 from 3 patients). From this study, the surveyed coastal area of Muan-gun, Chollanam-do was proven to be a new endemic focus of H. nocens and P. summa. The occurrence of a few infected cases suggests that this area should also be a low-grade endemic area of S. fuscata, H. continua, and G. seoi.


Cross talk between adhesion molecules: control of N-cadherin activity by intracellular signals elicited by beta1 and beta3 integrins in migrating neural crest cells.
During embryonic development, cell migration and cell differentiation are associated with dynamic modulations both in time and space of the repertoire and function of adhesion receptors, but the nature of the mechanisms responsible for their coordinated occurrence remains to be elucidated. Thus, migrating neural crest cells adhere to fibronectin in an integrin-dependent manner while maintaining reduced N-cadherin-mediated intercellular contacts. In the present study we provide evidence that, in these cells, the control of N-cadherin may rely directly on the activity of integrins involved in the process of cell motion. Prevention of neural crest cell migration using RGD peptides or antibodies to fibronectin and to beta1 and beta3 integrins caused rapid N-cadherin-mediated cell clustering. Restoration of stable intercellular contacts resulted essentially from the recruitment of an intracellular pool of N-cadherin molecules that accumulated into adherens junctions in tight association with the cytoskeleton and not from the redistribution of a preexisting pool of surface N-cadherin molecules. In addition, agents that cause elevation of intracellular Ca2+ after entry across the plasma membrane were potent inhibitors of cell aggregation and reduced the N-cadherin- mediated junctions in the cells. Finally, elevated serine/ threonine phosphorylation of catenins associated with N-cadherin accompanied the restoration of intercellular contacts. These results indicate that, in migrating neural crest cells, beta1 and beta3 integrins are at the origin of a cascade of signaling events that involve transmembrane Ca2+ fluxes, followed by activation of phosphatases and kinases, and that ultimately control the surface distribution and activity of N-cadherin. Such a direct coupling between adhesion receptors by means of intracellular signals may be significant for the coordinated interplay between cell-cell and cell-substratum adhesion that occurs during embryonic development, in wound healing, and during tumor invasion and metastasis.


Mating systems and protein-protein interactions determine evolutionary rates of primate sperm proteins.
To assess the relative impact of functional constraint and post-mating sexual selection on sequence evolution of reproductive proteins, we examined 169 primate sperm proteins. In order to recognize potential genome-wide trends, we additionally analysed a sample of altogether 318 non-reproductive (brain and postsynaptic) proteins. Based on cDNAs of eight primate species (Anthropoidea), we observed that pre-mating sperm proteins engaged in sperm composition and assembly show significantly lower incidence of site-specific positive selection and overall lower non-synonymous to synonymous substitution rates (dN/dS) across sites as compared with post-mating sperm proteins involved in capacitation, hyperactivation, acrosome reaction and fertilization. Moreover, database screening revealed overall more intracellular protein interaction partners in pre-mating than in post-mating sperm proteins. Finally, post-mating sperm proteins evolved at significantly higher evolutionary rates than pre-mating sperm and non-reproductive proteins on the branches to multi-male breeding species, while no such increase was observed on the branches to unimale and monogamous species. We conclude that less protein-protein interactions of post-mating sperm proteins account for lowered functional constraint, allowing for stronger impact of post-mating sexual selection, while the opposite holds true for pre-mating sperm proteins. This pattern is particularly strong in multi-male breeding species showing high female promiscuity.


Chemical etching in processing cortical bone specimens for scanning electron microscopy.
Transverse and longitudinal sectioning of undecalcified cortical bone is a commonly employed technique for investigating the lamellar structure of the osteons. Since a flat surface is required, the specimen has to be grinded and then polished. Whereas the smear of debris and inorganic/organic deposits left by these treatments cannot be removed by ultrasonication alone, a chemical treatment of the specimen surface with either a basic or an acid etching solution is currently employed. A further effect of the latter can be the enhancement of the lamellar bone pattern. The kind of etching solution, its pH, the concentration of etchants, and the contact time significantly affect the sectioned surface when it is observed with scanning electron microscopy (SEM). The etching procedures can severely influence the obtained images. Homogeneous cortical bone specimens were sampled from the first metatarsal of two fresh human subjects. One or two cut surfaces were exposed to different acid and basic solutions in bonded conditions. Considering the type of chemical agents, the solution pH, and the exposure time of the specimens, the effects of several etching media have been investigated and compared. Strong etching, either acid or basic produced surface decalcification and severe damage of the collagen matrix, compromising any morphological or morphometric analysis. Weak acid etching (for example citric and acetic acid), even though causing distinctive alteration of the sample, enhanced the visibility of the lamellar pattern, while the polyphosphate treatment of the surface decalcified a thin layer matrix, ensuring a good visibility of fibrils and avoiding rough distortions.


Striking Similarities Between Botrytis cinerea From Non-agricultural and From Agricultural Habitats.
Investigations into life history of microorganisms that cause plant diseases have been limited mostly to contexts where they are in interaction with plants, and with cropped or otherwise managed vegetation. Therefore, knowledge about the diversity of plant pathogens, about potential reservoirs of inoculum and about the processes that contribute to their survival and adaptation is limited to these contexts. The agro-centric perspective of plant pathogen life histories is incoherent with respect to the capacity of many of them to persist as saprophytes on various substrates. In this context we have investigated the ubiquity of the broad host range necrotrophic fungus Botrytis cinerea, outside of agricultural settings and have determined if the populations in these natural habitats can be distinguished phenotypically and phylogenetically from populations isolated from diseased crops. Over a period of 5 years, we isolated B. cinerea from 235 samples of various substrates collected in France including rainfall, snowpack, river, and lake water, epilithic biofilms in mountain streams, leaf litter and plant debris, rock surfaces, bird feathers and healthy wild plants from outside of agricultural fields. All substrates except rock surfaces harbored B. cinerea leading us to establish a collection of purified strains that were compared to B. cinerea from diseased tomato, grapes and various other crops in France. Phylogenetic comparisons of 321 strains from crop plants and 100 strains from environmental substrates based on sequences of 9 microsatellite markers revealed that strains from crops and the environment could not be distinguished. Furthermore, the genetic diversity of strains outside of agriculture was just as broad as within agriculture. In tests to determine the aggressiveness of strains on tomato stems, the mean disease severity caused by strains from environmental substrates was statistically identical to the severity of disease caused by strains from tomato, but was significantly greater than the severity caused by strains from grape or other crops. Our results suggest that highly diverse populations of this plant pathogen persist outside of agriculture in association with substrates other than plants and that this part of their life history is compatible with its capacity to maintain its potential as plant pathogen.


Objective elaluation of bypass surgery in patients with acute coronary artery disease.
80 patients with acute coronary artery disease, including 70 patients with PIA, 8 patients with acute myocardial infarction, and 2 patients with cardiogenic shock, underwent bypass surgery. The surgical mortality rate was 1.4% in patients with PIA and during an average follow-up period of 15 months none of the patients died of cardiac disease. Ventricular function, as indicated by systolic ejection fraction and segmental wall motion, was noted to improve in many patients. Postoperative studies showed the internal mammary artery to be superior to the saphenous vein graft for left coronary bypass procedures. This was demonstrated by 100% patency of internal mammary artery grafts to the left anterior descending coronary artery. We believe that when a patient presents with one of the syndromes of acute coronary artery disease such as unstable angina, severe chest pain suggestive of infarction without infarction, status anginosis, recurrent ventricular tachycardia or acute myocardial infarction complicated by evidence of potential extension, cardiogenic shock, heart block or rupture, these patients deserve at least coronary and left ventricular cineangiographic studies with hemodynamic evaluation. Our experience to date suggests that myocardial revascularization and catheterization carry a lesser risk than that of medical treatment.


Neurohypophysial hormone receptors and second messengers in trout hepatocytes.
Neurohypophysial hormone receptors and second messengers were studied in trout (Oncorhynchus mykiss) hepatocytes. Arginine vasotocin (AVT) and isotocin (IT) elicited a concentration-dependent inhibition of cAMP accumulation in the presence of 5x10(-8) M glucagon (maximal effect for 4.5x10(-7) M and 1.4x10(-7) M, half-maximal effect for 2.1x10(-8) M and 0.7x10(-8) M, AVT and IT respectively). The effect of glucagon was inhibited up to 90% by AVT and 80% by IT. While AVT inhibited (up to 50%) the basal cAMP production, IT had no such action. Specific V(1) or V(2) analogues (with reference to vasopressin in mammals) were used for pharmacological characterization of the type of neurohypophysial hormone receptor involved in this inhibition. The V(1) agonist [Phe(2), Orn(8)]-oxytocin inhibited the glucagon-stimulated cAMP production with a maximal effect for 6x10(-7) M and a half-maximal effect for 0.9x10(-8) M concentrations of the analogue. While the V(1) agonist reduced the glucagon-stimulated cAMP level by 70%, it showed only a tendency to reduce the basal level. The V(2) agonist [deamino(1), Val(4),d -Arg(8)]-vasopressin had no effect either on basal or on glucagon-stimulated cAMP production. The V(1) antagonist [d(CH(2))(5)(1), O-Me-Tyr(2), Arg(8)]-vasopressin totally reversed the 10(-8) M AVT-induced inhibition of 5x10(-8) M glucagon-stimulated cAMP production, whereas the V(2) antagonist [d(CH(2))(5)(1),d -Ile(2), Ile(4), Arg(8), Ala(9)]-vasopressin had no such effect. In this particular case, maximal and half-maximal effects of the V(1) antagonist were obtained for 2.3x10(-6) M and 1. 2x10(-6 )M respectively. Changes in intracellular calcium content were measured using the fluorescent probe FURA-2/AM. AVT and IT elicited a concentration-dependent increase in Ca(2+) accumulation. The comparison of the effect of 10(-8) M agonists versus AVT showed the following order of potency: AVT=IT>V(1) agonist>V(2) agonist. The V(1) antagonist reversed the AVT-induced Ca(2+) accumulation whereas the V(2) antagonist had no such effect. These results are taken as evidence for the presence in trout hepatocytes of neurohypophysial hormone receptors functionally close to the V(1a)-type linked to cAMP production and Ca(2+) mobilization.


Addressing Psychosocial Factors in Cognitive Impairment Screening from a Holistic Perspective: The DeCo-Booklet Methodology Design and Pilot Study.
Cognitive impairment (CI), an intermediate phase between the decline in physiological cognition and dementia, is known to be mediated by a variety of risk and protective factors, with age being the most influential of these. The multifactorial nature of CI and the worldwide phenomenon of an aging population makes decoupling old age from disease through the concept of healthy aging (HA) a matter of major interest. Focusing on psychosocial variables and psychological constructs, here we designed and piloted a data collection booklet (DeCo-B) to assess CI and HA from a holistic perspective. The DeCo-B comprises six sections: sociodemographic factors, CI, meaning in life, psychosocial factors, health problems, and lifestyle. The estimated prevalence of CI and HA in our cohort were 24.4% and 6.6%, respectively. Spearman correlations mainly identified pairwise associations between the meaning in life domains and psychosocial variables. Moreover, age, marital status, purpose in life, resilience, chronic pain, cognitive reserve, and obstructive sleep apnea were significantly associated with an increased risk of CI. Our results showed that DeCo-B is a suitable tool for researching how modifiable risk and protective factors influence cognitive status. The complex interrelationships between variables should be further investigated and, for practical reasons, the questionnaire should be optimized in future work.


[Origin and elimination of fibrous elements seen in CRL antirabies vaccine for human use].
The present work describes the discovery of an organoleptic alteration in some batchs of suckling mouse brain rabies vaccine for human use, produced in the Central Laboratory of Public Health of the Province of Buenos Aires. This alteration consists in the appearance of fibrous particles, light-white, slightly perceptible, suspended in the diluent. The production procedure is that of Fuenzalida and Palacios (4), with the modifications proposed by the Panamerican Zoonoses Center (Larghi and col.) (6), which include a refrigerated centrifugation of the viral suspension to 17,000 g during 10 minutes. This alteration caused 20% or more of dosis loss and a decrease in vaccine potency (2). In order to determine the origin of this alteration, we controlled the homogenization systems, the elements employed, the production of experimental batchs with diversity of isotonizators (dextrose-sucrose) and preservatives, the utilization of glass filters in the treatment of water, all drugs employed, variation of pH of diluent solutions, double centrifugation, the glass employed (ampoules, flasks), pumps of dispensation and wash systems (3) (5). We conclude that the alterations are produced by partial freezing of the infected brain suspension to 10% in water, during the last part of the centrifugation. Particles are formed by irreversible precipitation of a proteic nature (1) (7). It is recommended that during the production of this vaccine the temperature be maintained between 3 to 5 degrees C carefully avoiding freezing the brain suspension in distilled water.


Patient preferences and perspectives regarding reducing alcohol consumption: role of nalmefene.
Alcohol use disorder is a major public health issue. The absolute mortality burden of alcohol-attributable death has increased over the last 20 years. However, access to care remains very poor and many people with alcohol use disorder are untreated. The main limiting factor for access to care in alcohol use disorder appears to be the reluctance to engage in abstinence. Risk reduction is a developing approach in the treatment of alcohol use disorders, drawing its inspiration, with quite a delay, from the decades-long dominant approach in other substance use disorders. A paradigm shift has recently occurred that places more of an emphasis on reducing alcohol as a therapeutic strategy for patients with alcohol use disorder, to better meet the patients' preferences and needs. The development and recent approval of nalmefene, in alcohol-dependent adults with a high drinking risk level, contributes to enlarging the therapeutic arsenal for alcohol dependence, strengthening the legitimacy of alcohol reduction strategies.


[Value of central aortic pressure measurement in patients with arterial hypertension.].
The article is devoted to analysis of the effects of antihypertensive drugs on central aortic pressure in patients with arterial hypertension. Results of ASCOT-CAFcapital short I, Cyrillic trial are discussed. The importance of investigation of large arteries in arterial hypertension was noted.


The tunable hydrophobic effect on electrically doped graphene.
Using molecular dynamics simulations, we study the hydrophobic effect on electrically doped single layer graphene. With doping levels measured in volts, large changes in contact angle occur for modest voltages applied to the sheet. The effect can be understood as a renormalization of the surface tension between graphene and water in the presence of an electric field generated by the dopant charge, an entirely collective effect termed electrowetting. Because the electronic density of states scales linearly in the vicinity of the Fermi energy, the cosine of the contact angle scales quartically with the applied voltage rather than quadratically, as it would for a two-dimensional metal or in multiple layer graphene. While electrowetting explains the phenomenon, it does not account for the slight asymmetry observed in the hydrophobic response between n- and p-doping.


E2F7 Is a Potent Inhibitor of Liver Tumor Growth in Adult Mice.
Up-regulation of the E2F-dependent transcriptional network has been identified in nearly every human malignancy and is an important driver of tumorigenesis. Two members of the E2F family, E2F7 and E2F8, are potent repressors of E2F-dependent transcription. They are atypical in that they do not bind to dimerization partner proteins and are not controlled by retinoblastoma protein. The physiological relevance of E2F7 and E2F8 remains incompletely understood, largely because tools to manipulate their activity in vivo have been lacking. Here, we generated transgenic mice with doxycycline-controlled transcriptional activation of E2f7 and E2f8 and induced their expression during postnatal development, in adulthood, and in the context of cancer. Systemic induction of E2f7 and, to lesser extent, E2f8 transgenes in juvenile mice impaired cell proliferation, caused replication stress, DNA damage, and apoptosis, and inhibited animal growth. In adult mice, however, E2F7 and E2F8 induction was well tolerated, yet profoundly interfered with DNA replication, DNA integrity, and cell proliferation in diethylnitrosamine-induced liver tumors. Collectively, our findings demonstrate that atypical E2Fs can override cell-cycle entry and progression governed by other E2F family members and suggest that this property can be exploited to inhibit proliferation of neoplastic hepatocytes when growth and development have subsided during adulthood.


Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation.
PD-1 is an immunoinhibitory receptor expressed by activated T cells, B cells, and myeloid cells. Mice deficient in PD-1 exhibit a breakdown of peripheral tolerance and demonstrate multiple autoimmune features. We report here that the ligand of PD-1 (PD-L1) is a member of the B7 gene family. Engagement of PD-1 by PD-L1 leads to the inhibition of T cell receptor-mediated lymphocyte proliferation and cytokine secretion. In addition, PD-1 signaling can inhibit at least suboptimal levels of CD28-mediated costimulation. PD-L1 is expressed by antigen-presenting cells, including human peripheral blood monocytes stimulated with interferon gamma, and activated human and murine dendritic cells. In addition, PD-L1 is expressed in nonlymphoid tissues such as heart and lung. The relative levels of inhibitory PD-L1 and costimulatory B7-1/B7-2 signals on antigen-presenting cells may determine the extent of T cell activation and consequently the threshold between tolerance and autoimmunity. PD-L1 expression on nonlymphoid tissues and its potential interaction with PD-1 may subsequently determine the extent of immune responses at sites of inflammation.


Functional differences between two classes of sodium channels in developing rat skeletal muscle.
Excitability is generated in developing skeletal muscle by the incorporation of sodium-selective ion channels into the surface membrane. Whole-cell and patch voltage-clamp recording from myotubes and their embryologic precursors, myoblasts, indicated that voltage-activated sodium current in myoblasts was more resistant to block by tetrodotoxin (TTX) than that in myotubes. Single-channel recording from both cell types showed two classes of sodium channels. One class had a lower single-channel conductance, activated at more hyperpolarized voltages, and was more resistant to TTX than the other. The proportion of TTX-resistant to TTX-sensitive sodium channels was higher in myoblasts than in myotubes. Thus, the difference in TTX sensitivity between myoblasts and myotubes can be explained by a difference in the proportion of the two classes of sodium channels. In addition, the lower conductance of TTX-resistant channels provides insight into the relationship between the TTX binding site and the external mouth of the sodium channel.


[Stroke epidemiology and evaluation of risk factors significance for survival of patients during 28 poststroke days in the open population of Chita].
To provide effective primary and secondary stroke prevention, the epidemiological stroke rates were investigated in the open population of a large Transbaykal town by the register method. During one year, 989 cases have been examined in the population aged above 25 years. The age-standardized stroke incidence rate (per 100 000 persons) was 300 in women and 329 in men. The age-standardized stroke morbidity (per 100 000 persons) was 390 in women and 409 in men. Stroke mortality was 77 in women and 81 in men per 100 000 (p = 0.01). The stroke mortality in men was higher than in women. Stroke case fatality was 20% in men and 16% in women. The prevalence and significance of the main cardiovascular risk factors were revealed among patients by case-control study. Hypertension, psychological and social risk factors were the most significant in Chita's population of stroke patients.


Individualization and the Health Care Mosaic in Assisted Living.
Assisted living (AL) is a popular residential long-term care option for frail older adults in the United States. Most residents have multiple comorbidities and considerable health care needs, but little is known about their health care arrangements, particularly over time. Our goal is to understand how health care is managed and experienced in AL by residents and their care network members. This grounded theory analysis focuses on the delivery of health care in AL. Qualitative data were gathered from 28 residents and 114 of their care network members followed over a 2-year period in 4 diverse settings as part of the larger study, "Convoys of Care: Developing Collaborative Care Partnerships in Assisted Living." Findings show that health care in AL involves routine, acute, rehabilitative, and end-of-life care, is provided by residents, formal and informal caregivers, and occurs on- and off-site. Our conceptual model derived from grounded theory analysis, "individualizing health care," reflects the variability found in care arrangements over time and the multiple, multilevel factors we identified related to residents and caregivers (e.g., age, health), care networks (e.g., size, composition), residences (e.g., ownership), and community and regulatory contexts. This variability leads to individualization and a mosaic of health care among AL residents and communities. Our consideration of health care and emphasis on care networks draw attention to the importance of communication and need for collaboration within care networks as key avenues for improving care for this and other frail populations.


Do static and dynamic insulin resistance indices perform similarly in predicting pre-diabetes and type 2 diabetes?
We designed a study to compare the predictive power of static and dynamic insulin resistance indices for categorized pre-diabetes (PDM)/type 2 diabetes (DM). Participants included 1134 adults aged 18-60 years old with normal glucose at baseline who completed both baseline and 6-years later follow-up surveys. Insulin resistance indices from baseline data were used to predict risk of PDM or DM at follow-up. Two static indices and two dynamic indices were calculated from oral glucose tolerance test results (OGTT) at baseline. Area under the receiver operating characteristic curve (AROC) analysis was used to estimate the predictive ability of candidate indices to predict PDM/DM. A general estimation equation (GEE) model was applied to assess the magnitude of association of each index at baseline with the risk of PDM/DM at follow-up. The dynamic indices displayed the largest and statistically predictive AROC for PDM/DM diagnosed either by fasting glucose or by postprandial glucose. The bottom quartiles of the dynamic indices were associated with an elevated risk of PDM/DM vs. the top three quartiles. However, the static indices only performed significantly to PDM/DM diagnosed by fasting glucose. Dynamic insulin resistance indices are stronger predictors of future PDM/DM than static indices. This may be because dynamic indices better reflect the full range of physiologic disturbances in PDM/DM.


Germline Variants in the POT1-Gene in High-Risk Melanoma Patients in Austria.
Risk of melanoma is in part determined by genetic factors. Currently the only established high penetrance familial melanoma genes are CDKN2A and CDK4. Recent studies reported germline variants in POT1 in melanoma families. In the present study, we sequenced the entire POT1 gene in 694 patients from the M3-study. Patients with multiple primary melanomas (n = 163) or with a positive family history (n = 133) were classified as high-risk melanoma patients. Additionally, 200 single primary melanoma patients and 198 non-melanoma controls were sequenced. For prediction analysis 10 different tools were used.In total 53 different variants were found, of which 8 were detected in high-risk melanoma patients, only. Two out of these 8 variants were located in exons and were non-synonymous: g.124510982 G>A (p.R80C) and g.124491977 T>G (p.N300H). While g.124491977 T>G was predicted to be neutral, 80% of the prediction tools classified g.124510982 G>A as deleterious. The variant, g.124467236 T>C, which possibly causes a change in the splice site was identified in a case with a positive family history in the present study. Another variant in the 5-UTR, g.124537261 A>G, was found in 2 high-risk patients. So, in conclusion, melanoma associated POT1 germline variants seem to be rare. Further studies are required to evaluate the role of POT1 for genetic counseling.


Food addiction: A common neurobiological mechanism with drug abuse.
Drugs and food both exert a rewarding effect through the firing of dopamine neurons in the ventral tegmental area, resulting in the release of dopamine into the nucleus accumbens and effects on the mesolimbic pathway. Here, we review the neuroimaging literature to consider the validity of food addiction and the common neurobiological mechanisms that overlap in food and drug addiction. This review paper focuses on findings from Positron Emission Tomography (PET), functional Magnetic Resonance Imaging (fMRI) and structural imaging studies, as well as evidence from neuroimaging studies of bariatric surgery and pharmacological interventions on obese individuals. We examine not only functional and structural changes in the mesolimbic pathways, but also in other frontal areas shown to be involved in drug addiction, including the prefrontal cortex, orbitofrontal cortex and anterior cingulate cortex, as well as changes in neurotransmitter systems beyond dopaminergic systems.


Plasma membrane targetting, vesicular budding and release of galectin 3 from the cytoplasm of mammalian cells during secretion.
Galectin 3, a 30 kDa galactoside-binding protein distributed widely in epithelial and immune cells, contains no signal sequence and is externalized by a mechanism independent of the endoplasmic reticulum (ER)-Golgi complex. We show here that hamster galectin 3 overexpressed in transfected cos-7 cells is secreted at a very low rate. A chimaera of galectin 3 fused to the N-terminal acylation sequence of protein tyrosine kinase p56(lck), Nt-p56(lck)-galectin 3, which is myristoylated and palmitoylated and rapidly transported to plasma membrane domains, is efficiently released from transfected cells indicating that movement of cytoplasmic galectin 3 to plasma membrane domains is a rate limiting step in lectin secretion. N-terminal acylation is not sufficient for protein secretion since p56(lck) and the chimaera Nt-p56(lck)-CAT are not secreted from transfected cells. The amino-terminal half of galectin 3 is sufficient to direct export of a chimaeric CAT protein indicating that part of the signal for plasma membrane translocation lies in the N-terminal domains of the lectin. Immunofluorescence studies show that Nt-p56(lck)-galectin 3 aggregates underneath the plasma membrane and is released by membrane blebbing. Vesicles of low buoyant density isolated from conditioned medium are enriched in galectin 3. The lectin is initially protected from exogenous collagenase but is later released in soluble protease-sensitive form from the lectin-loaded vesicles. Using murine macrophages, which secrete their endogenous galectin 3 at a moderate rate especially in the presence of Ca2+-ionophores, we were also able to trap a galectin 3-loaded vesicular fraction which was released into the culture supernatant.


Genetic variation in the retinoid X receptor and calcium-sensing receptor and risk of colorectal cancer in the Colon Cancer Family Registry.
Genetic variants in the calcium/vitamin D metabolic pathway may be related to risk for colorectal cancer. While several investigations of vitamin D receptor (VDR) polymorphisms and colorectal cancer have been conducted, no studies to date have evaluated the association of genetic variation in the heterodimer partner for VDR, the retinoid X receptor (RXR). Another important gene in this pathway is the calcium-sensing receptor (CASR). Employing a discordant-sibship case-control design, we examined the association between single nucleotide polymorphisms (SNPs) in RXRA and CASR and risk for colorectal cancer overall and by colorectal subsite and microsatellite instability (MSI) status using data from the Colon Cancer Family Registry. No gene-level relationships between RXRA or CASR and colorectal cancer overall were observed. However, for RXRA SNP rs7861779, a high-interest SNP selected for study a priori, there was a statistically significantly increased risk for proximal colorectal cancer among those with at least one A allele [odds ratio (OR) = 1.42; 95% confidence interval (CI) = 1.03-1.97]. Another selected RXRA SNP, rs12004589, was significantly associated with risk of MSI-high cancers (OR = 2.27; 95% CI = 1.13-4.56). Additionally, CASR SNP rs1801726 was significantly associated with a reduced risk for rectal cancer (OR = 0.53; 95% CI = 0.29-0.96). These results provide support that RXRA SNPs rs7861779 and rs12004589 and CASR SNP rs1801726 may be important markers for colorectal neoplasia. Further work is needed to elucidate their role in the carcinogenic pathway.


ANCA-negative limited Wegener's granulomatosis.
A 26-year-old man presented with epistaxis, nasal obstruction and a subcutaneous swelling over the left malar region with radiological evidence of a mass in the right nasal cavity. Histology of the lesions showed necrotizing granuloma with evidence of vasculitis. There was no other systemic involvement and the patient was ANCA-negative. Excellent response to systemic steroid and cyclophosphamide therapy was noted.


Trefoil factor family peptides enhance cell migration by increasing cellular osmotic permeability and aquaporin 3 levels.
Trefoil factor family (TFF) peptides are produced rapidly at sites of injury, stimulating epithelial migration, a process involving rapid changes in cell shape and volume, requiring rapid flow of water into and out of the cell. We examined the effect of TFFs on fluidity of cells by measuring their sensitivity to osmotic challenges and cell migration, and determined whether those results were mediated through altering the levels of aquaporins (AQPs), a family of transmembrane water channels involved in cellular water homeostasis. Gastric (AGS) and colonic (Caco-2) cell lines had intrinsic TFF levels determined and the predominant TFF peptide knocked down (RNA interference). Knockdown caused lessened responsiveness to changes in external osmotic challenge (by 51 and 69% in AGS and Caco-2 cells, respectively) and reduced cell migration and transepithelial permeability but did not influence proliferation. Exogenous TFF increased several AQPs, particularly AQP3, and those were reciprocally reduced in knockdown cells. TFF-induced, but not fetal calf serum-induced, cell migration was inhibited by the presence of AQP3 blocker (CuSO4). We summarize that TFF peptides promptly produced at sites of injury increase AQP levels, most notably AQP3, thereby enhancing the cells' ability to rapidly change their shape as part of the restitutive process. TFF peptides also require functioning AQP3 channels to induce cell migration.-Marchbank, T., Playford, R. J. Trefoil factor family peptides enhance cell migration by increasing cellular osmotic permeability and aquaporin 3 levels.


Defining Recovery From Alcohol Use Disorder: Development of an NIAAA Research Definition.
The objective of this article is to provide an operational definition of recovery from alcohol use disorder (AUD) to facilitate the consistency of research on recovery and stimulate further research. The construct of recovery has been difficult to operationalize in the alcohol treatment and recovery literature. Several formal definitions of recovery have been developed but have limitations because 1) they require abstinence from both alcohol and substance use, 2) they do not include the DSM-5 diagnostic criteria for AUD as part of the recovery process (i.e., no focus on remission from AUD), 3) they do not link remission and cessation from heavy drinking to improvements in biopsychosocial functioning and quality-of-life constructs, and 4) they do not distinguish between alcohol and other drug use. The authors present a newly developed National Institute on Alcohol Abuse and Alcoholism (NIAAA) definition of recovery from DSM-5 AUD based on qualitative feedback from key recovery stakeholders (e.g., researchers, clinicians, and recovery specialists). The definition views recovery as both a process of behavioral change and an outcome and incorporates two key components of recovery, namely, remission from DSM-5 AUD and cessation from heavy drinking, a nonabstinent recovery outcome. The NIAAA definition of recovery also emphasizes the importance of biopsychosocial functioning and quality of life in enhancing recovery outcomes. This new NIAAA definition of recovery is an operational definition that can be used by diverse stakeholders to increase consistency in recovery measurement, stimulate research to better understand recovery, and facilitate the process of recovery.


Expression of slow skeletal troponin I in hearts of phospholamban knockout mice alters the relaxant effect of beta-adrenergic stimulation.
Beta-adrenergic stimulation of the heart results in an enhanced relaxation rate in association with phosphorylation of both cardiac troponin I (cTnI) and phospholamban (PLB). We studied new lines of mice generated by crossbreeding mice that express slow skeletal troponin I (ssTnI) with PLB knockout (PLBKO) mice. This crossbreeding resulted in the generation of PLB/cTnI, PLB/ssTnI, PLBKO/cTnI, and PLBKO/ssTnI mice. Perfusion with isoproterenol (ISO) significantly increased the peak amplitude of fura-2 ratio in PLB/cTnI, PLBKO/cTnI, and PLBKO/ssTnI groups of mice. However, in the presence of ISO, there were no differences in the peak amplitude of fura-2 ratio among cells isolated from hearts of PLB/cTnI, PLBKO/cTnI, and PLBKO/ssTnI mice. In cells from PLB/cTnI mice, the extent of shortening was increased and the time of relaxation was significantly decreased during beta-adrenergic stimulation. In PLBKO/cTnI cells, stimulation with ISO resulted in an increased extent of shortening and no change in time of relaxation. However, stimulation with ISO in cells isolated from PLBKO/ssTnI mice not only significantly increased the extent of cell shortening but also increased the time of relaxation. We also determined the kinetics of relaxation of papillary muscles isolated from all four groups of animals in the presence and absence of ISO. Perfusion with ISO increased the rate of relaxation only in PLB/cTnI, PLB/ssTnI, and PLBKO/cTnI muscles. During ISO stimulation, the time of relaxation was unchanged in PLBKO/ssTnI muscles. Our data directly demonstrate that phosphorylation of both PLB and cTnI contributes to increased rate of relaxation during beta-adrenergic stimulation.


Thoracoscopic placement of phrenic nerve electrodes for diaphragmatic pacing in children.
Diaphragmatic pacing can provide chronic ventilatory support for children who suffer from congenital central hypoventilation syndrome (CCHS) or cervical spinal cord injury. The authors report a new thoracoscopic approach for establishing diaphragm pacing. Between 1997 and 2000, 9 children ranging in age from 5 to 15 years and suffering from these disorders underwent thoracoscopic placement of bilateral phrenic nerve electrodes. A 3- or 4-trocar technique was used to dissect the phrenic nerve in the midchest and suture a phrenic nerve electrode (Avery Laboratories I-110A, Commack, NY) into place. The electrode was tunneled to a subcutaneous pocket in the upper abdomen and attached to an implanted pacing unit. Bilateral electrodes were placed successfully into all patients. The average procedure time was 3.3 hours (range, 2.5 to 4.6), and average hospital stay was 4.2 days (range, 3 to 5). Four patients experienced postoperative complications (pneumonia, atelectasis, bradycardia, and pneumothorax). Average follow-up has been 30 months (range, 15 to 49). Eight patients have reached their long-term pacing goals. Phrenic nerve electrodes can be implanted thoracoscopically and allow the successful use of diaphragmatic pacing therapy. Avoidance of thoracotomy with its associated perioperative morbidity and scarring may encourage wider utilization of diaphragmatic pacing in children.


Reducing needle stick injuries in healthcare occupations: an integrative review of the literature.
Needlestick injuries frequently occur among healthcare workers, introducing high risk of bloodborne pathogen infection for surgeons, assistants, and nurses. This systematic review aims to explore the impact of both educational training and safeguard interventions to reduce needlestick injuries. Several databases were searched including MEDLINE, PsycINFO, SCOPUS, CINAHL and Sciencedirect. Studies were selected if the intervention contained a study group and a control group and were published between 2000 and 2010. Of the fourteen studies reviewed, nine evaluated a double-gloving method, one evaluated the effectiveness of blunt needle, and one evaluated a bloodborne pathogen educational training program. Ten studies reported an overall reduction in glove perforations for the intervention group. In conclusion, this review suggests that both safeguard interventions and educational training programs are effective in reducing the risk of having needlestick injuries. However, more studies using a combination of both safeguards and educational interventions in surgical and nonsurgical settings are needed.


The human set and transposase domain protein Metnase interacts with DNA Ligase IV and enhances the efficiency and accuracy of non-homologous end-joining.
Transposase domain proteins mediate DNA movement from one location in the genome to another in lower organisms. However, in human cells such DNA mobility would be deleterious, and therefore the vast majority of transposase-related sequences in humans are pseudogenes. We recently isolated and characterized a SET and transposase domain protein termed Metnase that promotes DNA double-strand break (DSB) repair by non-homologous end-joining (NHEJ). Both the SET and transposase domain were required for its NHEJ activity. In this study we found that Metnase interacts with DNA Ligase IV, an important component of the classical NHEJ pathway. We investigated whether Metnase had structural requirements of the free DNA ends for NHEJ repair, and found that Metnase assists in joining all types of free DNA ends equally well. Metnase also prevents long deletions from processing of the free DNA ends, and improves the accuracy of NHEJ. Metnase levels correlate with the speed of disappearance of gamma-H2Ax sites after ionizing radiation. However, Metnase has little effect on homologous recombination repair of a single DSB. Altogether, these results fit a model where Metnase plays a role in the fate of free DNA ends during NHEJ repair of DSBs.


U.S. veterans' experiences and factors associated with use of a smartphone application to self-manage unhealthy alcohol use.
Unhealthy alcohol use is common among Operations Enduring and Iraqi Freedom (OEF/OIF) veterans, yet barriers discourage treatment-seeking. Mobile applications (apps) that deliver alcohol interventions have potential to address these barriers and increase treatment receipt. Few studies have qualitatively assessed users' experiences with apps to manage alcohol use. We assessed OEF/OIF veterans' experiences with Step Away, an app to reduce alcohol-related risks, to identify factors that may influence engagement. This single-arm pilot study recruited OEF/OIF veterans with positive alcohol screens nationwide using mail/telephone. Veterans aged 18-55 who exceeded drinking guidelines and owned an iPhone were eligible. Twenty-one (16 men, 5 women) of 55 participants completed interviews. Interviews were analyzed using thematic analysis. Participants found Step Away easy to use, although setup was time consuming. Participants reported increased awareness of alcohol use, highlighting daily assessment, weekly feedback, goal setting, and high-risk notification features as helpful and associated awareness with an intent to decrease use. Participants described Step Away as informative, with over half reporting they would use it outside of the study and most recommending it. Suggestions for improvement included greater personalization and control over features. Step Away features appear to influence engagement and increase users' awareness about alcohol consumed and factors associated with drinking, as well as intent to change. Assessment, feedback, and customization features of apps may facilitate app engagement. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


The role of wet wipes and sanitary towels as a source of white microplastic fibres in the marine environment.
Understanding source elements of the ocean plastic crisis is key to effective pollution reduction management and policy. The ubiquity of microplastic (MP) fibres in the oceans is considered to derive primarily from clothing fibres released in grey water. Microplastic fibres degraded from widely flushed personal care textile products (wet wipes and sanitary towels) have not been clearly identified in aquatic systems to date. Unregulated personal hygiene and sanitary product labelling fails to identify textile materials. This study demonstrated that white MP fibres in sediments adjacent to a wastewater treatment plant (WWTP) are comparable with white fibres from sewage-related waste and commercially available consumer sanitary products. Commercially available non-flushable wipes are manufactured from either polyethylene terephthalate (PET), polypropylene (PP), or a combination of PET and cellulose. Fifty percent of brands labelled flushable that were tested were comprised of a mixture of PET and cellulose and the remainder of cellulose alone. Sanitary towels are made from PP, PE, or a combination of high-density polyethylene (HDPE) and PP. The accumulation of large quantities of washed-up sewage-related macro-debris (including wet wipes and sanitary towels) intermingled with seaweed biomass adjacent to the WWTP was associated with a combined sewer overflow. Microplastic fibres extracted from this waste were similar to those extracted from intertidal sediments in close proximity to the WWTP over a ten-month period. In comparison, fibres extracted from locations spatially removed from the WWTP were primarily comprised of ABS, PP and polystyrene. The results confirm that wet wipes and sanitary towels flushed down toilets are an underestimated source of white MP fibres in the environment. Given the global distribution and projected growth of the non-woven textile industry, there is a need for increased public awareness of MP pollution in the marine environment from the inappropriate disposal of sanitary products down the toilet, instead of diversion to alternative land-based waste management.


Entorhinal and Transentorhinal Atrophy in Preclinical Alzheimer's Disease.
This study examines the atrophy patterns in the entorhinal and transentorhinal cortices of subjects that converted from normal cognition to mild cognitive impairment. The regions were manually segmented from 3T MRI, then corrected for variability in boundary definition over time using an automated approach called longitudinal diffeomorphometry. Cortical thickness was calculated by deforming the gray matter-white matter boundary surface to the pial surface using an approach called normal geodesic flow. The surface was parcellated based on four atlases using large deformation diffeomorphic metric mapping. Average cortical thickness was calculated for (1) manually-defined entorhinal cortex, and (2) manually-defined transentorhinal cortex. Group-wise difference analysis was applied to determine where atrophy occurred, and change point analysis was applied to determine when atrophy started to occur. The results showed that by the time a diagnosis of mild cognitive impairment is made, the transentorhinal cortex and entorhinal cortex was up to 0.6 mm thinner than a control with normal cognition. A change point in atrophy rate was detected in the transentorhinal cortex 9-14 years prior to a diagnosis of mild cognitive impairment, and in the entorhinal cortex 8-11 years prior. The findings are consistent with autopsy findings that demonstrate neuronal changes in the transentorhinal cortex before the entorhinal cortex.


Biliary hyperkinesia in adolescents-it isn't all hype!
Biliary dyskinesia generally refers to a hypofunctioning gallbladder with an ejection fraction (EF) of <35% on hepatobiliary iminodiacetic acid scan with cholecystokinin stimulation (CCK-HIDA testing). In adults, biliary hyperkinesia has a defined association with biliary colic symptoms and can be relieved with surgical intervention. This clinical entity has not been well described in children or adolescents. In fact, only recently have we seen biliary hyperkinesia on HIDA at our centers. To that end, we reviewed our recent experience with adolescents who have presented and been treated for this unusual clinical entity. With IRB approval, we retrospectively reviewed the records of all patients with abnormally high HIDA EFs (>80%) cared for by the pediatric surgery services at two tertiary care centers over the span of a three-year period. Age, sex, BMI, CCK-HIDA results, and preoperative testing and post-operative pathology were noted. Resolution of symptoms was determined by subjective patient self-reporting at postoperative visit. Eighteen patients met inclusion criteria. Average age 15.7 (range, 10-17 years), median BMI 27.3 (±8.2). Fifteen patients were female and 3 were male. Average CCK-HIDA EF was 91.6% (±5.2), 82.4% of the patients had evidence of chronic cholecystitis and/or cholesterolosis on pathology. Postoperatively, 82.4% of the patients available for follow up (n=17) reported complete or near complete resolution of symptoms. Biliary hyperkinesia is an emerging clinical entity in children and adolescents and has a similar presentation to biliary hypokinesia. While the pathophysiologic mechanism of pain is not fully elucidated, laparoscopic cholecystectomy appears to provide a surgical cure for these patients and should be considered in the differential for the patient with an unremarkable workup and history suggestive of biliary colic.


Mental health of parents having children with physical disabilities.
Parents of children with disabilities play an essential role in the successful rehabilitation of their children. However, the high level of care required may affect the mental health of the parents and consequently contribute to an unfavorable rehabilitation outcome in their children. The aims of this study are to investigate the mental health of parents of children with physical disabilities and to elucidate the factors affecting parental mental health. This was a cross-sectional study. Ninety-one parents of children with physical disabilities who visited the outpatient rehabilitation clinics of a tertiary hospital were invited to join the study. Data from the parents and children were recorded during face-to-face interviews. Parental- perceived stress was assessed using the simplified Parenting Stress Index (PSI/SF). Parental mental health was evaluated as the main outcome measure using the 12-item version of the Chinese Health Questionnaire (CHQ-12). Forty parents (44%) were categorized as psychiatric cases on the CHQ. Child-related factors that negatively affected parental mental health were poor walking ability (p < 0.05), dependency on others to perform activities of daily life (ADL) (p < 0.01), and younger age (p < 0.05). Parent-related factors that negatively affected parental mental health were low income (p < 0.05), no religious beliefs (p < 0.01), high level of parental distress (p < 0.01), parent-child dysfunctional interaction (p < 0.01), and having difficult child (p < 0.05). The most significant predictor of overall parental mental health in multivariate analysis was found to be parental distress. Parents of children with physical disabilities were at risk of poor mental health. Perceived parental distress is the most important factor affecting parental mental health. Strategies to reduce parental stress should be developed to prevent deterioration of parental mental health.


Secondhand smoke drift: examining the influence of indoor smoking bans on indoor and outdoor air quality at pubs and bars.
This study aimed to examine the influence of indoor smoking bans on indoor and outdoor air quality at pubs and bars and to assess whether secondhand tobacco smoke (SHS) drifts from outdoor smoking areas to adjacent indoor areas. Data were covertly collected from a convenience sample of 19 pubs and bars that had at least 1 indoor area with an adjacent semi-enclosed outdoor eating/drinking area. Using TSI SidePak Personal Aerosol Monitors, concentrations of SHS (PM(2.5)) were measured concurrently in indoor and outdoor areas before and after implementation of the indoor smoking ban. Information was collected about the number of patrons and lit cigarettes and about the enclosure of outdoor areas. Indoor PM(2.5) concentrations reduced by 65.5% from pre-ban to post-ban (95% CI 32.6%-82.3%, p = .004). Outdoor exposure to PM(2.5) also reduced from pre-ban to post-ban by 38.8% (95% CI 3.2%-61.3%, p = .037). At post-ban, indoor concentrations of PM(2.5) were positively associated with outdoor concentrations. After adjustment for covariates, a 100% increase in geometric mean (GM) outdoor PM(2.5) was associated with a 36.1% (95% CI 2.4%-80.9%) increase in GM indoor PM(2.5) exposure (p = .035). Indoor smoking bans are an effective means of improving indoor and outdoor air quality in pubs and bars, although the air quality of smoke-free indoor areas may be compromised by smoking in adjacent outdoor areas. These findings require consideration in efforts to ensure adequate protection of the health of employees and patrons at hospitality venues.


Switches and latches: a biochemical tug-of-war between the kinases and phosphatases that control mitosis.
Activation of the cyclin-dependent kinase (Cdk1) cyclin B (CycB) complex (Cdk1:CycB) in mitosis brings about a remarkable extent of protein phosphorylation. Cdk1:CycB activation is switch-like, controlled by two auto-amplification loops--Cdk1:CycB activates its activating phosphatase, Cdc25, and inhibits its inhibiting kinase, Wee1. Recent experimental evidence suggests that parallel to Cdk1:CycB activation during mitosis, there is inhibition of its counteracting phosphatase activity. We argue that the downregulation of the phosphatase is not just a simple latch that suppresses futile cycles of phosphorylation/dephosphorylation during mitosis. Instead, we propose that phosphatase regulation creates coherent feed-forward loops and adds extra amplification loops to the Cdk1:CycB regulatory network, thus forming an integral part of the mitotic switch. These network motifs further strengthen the bistable characteristic of the mitotic switch, which is based on the antagonistic interaction of two groups of proteins: M-phase promoting factors (Cdk1:CycB, Cdc25, Greatwall and Endosulfine/Arpp19) and interphase promoting factors (Wee1, PP2A-B55 and a Greatwall counteracting phosphatase, probably PP1). The bistable character of the switch implies the existence of a CycB threshold for entry into mitosis. The end of G2 phase is determined by the point where CycB level crosses the CycB threshold for Cdk1 activation.


Instrumented facet fusion for the degenerative lumbar disorders.
A new, simple, minimally morbid procedure used to treat degenerative lumbar spinal disorders is described. The authors based their treatment on the McBride technique of facet fusion, which was modified and supplemented with pedicle screw fixation. The first 32 consecutive patients with degenerative spondylolisthesis and failed back surgery syndrome were treated, and followed for more than a year. Surgically, 26 patients had a single level fusion, and 6 patients had a two level fusion. Postoperative radiographs and computed tomography (CT) scans were evaluated to determine the fusion status. There were no specific complications related with the facet fusion. Thirty of 32 (93.8%) had solid fusions according to the computed tomographic criteria. Two patients showed a questionable union on CT scan but motion was less than 5 degrees on dynamic films. While 96.2% (25/26) of patients with a single level procedure had solid fusion, rate of union in the patients with two level procedures was 83.3% (5/6). This preliminary study of 32 patients shows that instrumented facet fusion appears to be a safe and effective procedure for lumbar spine fusions, demonstrating a high fusion rate with rarely serious complications.