Interpreting a black box predictor to gain insights into early folding mechanisms.
Protein folding and function are closely connected, but the exact mechanisms by which proteins fold remain elusive. Early folding residues (EFRs) are amino acids within a particular protein that induce the very first stages of the folding process. High-resolution EFR data are only available for few proteins, which has previously enabled the training of a protein sequence-based machine learning 'black box' predictor (EFoldMine). Such a black box approach does not allow a direct extraction of the 'early folding rules' embedded in the protein sequence, whilst such interpretation is essential to improve our understanding of how the folding process works. We here apply and investigate a novel 'grey box' approach to the prediction of EFRs from protein sequence to gain mechanistic residue-level insights into the sequence determinants of EFRs in proteins. We interpret the rule set for three datasets, a default set comprised of natural proteins, a scrambled set comprised of the scrambled default set sequences, and a set of de novo designed proteins. Finally, we relate these data to the secondary structure adopted in the folded protein and provide all information online via http://xefoldmine.bio2byte.be/, as a resource to help understand and steer early protein folding.


Autofluorescence: a screening test for mycotic infection in tissues.
Fungal infection is a major health concern as the clinical features are not very distinctive. Lack of rapid diagnostic techniques results in delay in diagnosis, which may even culminate in a fatal outcome. The fact that many pathogenic fungal organisms autofluoresce in hematoxylin and eosin (H and E)-stained sections under ultraviolet illumination led us to evaluate the role of autofluorescence as a rapid screening technique for fungal infections. The aim of the present study was to assess the value of autofluorescence as a screening method for detecting fungi on tissue sections and to compare the results of autofluorescence with conventional histochemical stains for fungi. Hematoxylin and eosin-stained slides of mycotic lesions were examined under fluorescent microscope and the findings were compared with results of Gomori's methenamine silver and periodic acid-Schiff stains. We found fungal autofluorescence in 63 out of 64 cases studied, with a sensitivity of 97.8% and specificity of 100% in comparison with fungal stains. This was statistically significant (P < 0.05). We conclude that autofluorescence can be used as a rapid screening method for identification of fungi in tissue sections as it does not require any other specialized staining procedure.


Crystal structure of the catalytic subunit of Cdc25B required for G2/M phase transition of the cell cycle.
Cdc25B is a dual specificity phosphatase involved in the control of cyclin-dependent kinases and the progression of cells through the cell cycle. A series of minimal domain Cdc25B constructs maintaining catalytic activity have been expressed. The structure of a minimum domain construct binding sulfate was determined at 1.9 A resolution and a temperature of 100 K. Other forms of the same co?nstruct were determined at lower resolution and room temperature. The overall folding and structure of the domain is similar to that found for Cdc25A. An important difference between the two is that the Cdc25B domain binds oxyanions in the catalytic site while that of Cdc25A appears unable to bind oxyanions. There are also important conformational differences in the C-terminal region. In Cdc25B, both sulfate and tungstate anions are shown to bind in the catalytic site containing the signature motif (HCxxxxxR) in a conformation similar to that of other protein tyrosine phosphatases and dual specificity phosphatases, with the exception of the Cdc25A. The Cdc25B constructs, with various truncations of the C-terminal residues, are shown to have potent catalytic activity. When cut back to the site at which the Cdc25A structure begins to deviate from the Cdc25B structure, the activity is considerably less. There is a pocket extending from the catalytic site to an anion-binding site containing a chloride about 14 A away. The catalytic cysteine residue, Cys473, can be oxidized to form a disulfide linkage to Cys426. A readily modifiable cysteine residue, Cys484, resides in another pocket that binds a sulfate but not in the signature motif conformation. This region of the structure is highly conserved between the Cdc25 molecules and could serve some unknown function.


MetProc: Separating Measurement Artifacts from True Metabolites in an Untargeted Metabolomics Experiment.
High-throughput metabolomics using liquid chromatography and mass spectrometry (LC/MS) provides a useful method to identify biomarkers of disease and explore biological systems. However, the majority of metabolic features detected from untargeted metabolomics experiments have unknown ion signatures, making it critical that data should be thoroughly quality controlled to avoid analyzing false signals. Here, we present a postalignment method relying on intermittent pooled study samples to separate genuine metabolic features from potential measurement artifacts. We apply the method to lipid metabolite data from the PREDIMED (PREvención con DIeta MEDi-terránea) study to demonstrate clear removal of measurement artifacts. The method is publicly available as the R package MetProc, available on CRAN under the GPL-v2 license.


Is there a role for the immune system in glaucomatous optic neuropathy?
Glaucoma and immunity are not traditionally perceived as being causally related. Recently, however, compelling observations have provided insight into a potential role for the immune system in the development of glaucomatous optic neuropathy. In this article, it is proposed that the role of the immune system in glaucoma is two-fold. In some patients, there is evidence that an autoimmune mechanism may be responsible for eliciting damage to the optic nerve, resulting in glaucomatous injury. Autoimmune damage to the optic nerve may occur directly by autoantibodies, or indirectly by way of a "mimicked" autoimmune response to a sensitizing antigen which, in turn, injuries retinal ganglion cells. We suggest that autoimmune-mediated glaucoma injury occurs most often, but not exclusively, in patients in whom the intraocular pressure has never been found to be elevated. A second role of the immune system in glaucoma is likely one of surveillance, in which signal pathways of the immune system regulate cell death in response to conditions that stress retinal neurons in glaucoma. These might include mechanical stress from high intraocular pressure, ischemia, excessive excitatory amino acids, or toxic products resulting from excessive nitric oxide synthase production in either neurons or glial fibers that surround the optic nerve as it exits the eye. In these cases, we propose that the immune system acts as an "arbiter" to help determine whether a neuronal cell will ultimately survive, or succumb to, those stressors that are perceived as injurious. It is conceivable that such surveillance and cell death regulation by the immune system is important in determining the fate of retinal neurons in both the more common "high-pressure" forms of glaucoma, such as primary open-angle glaucoma, and in cases in which the intraocular pressure appears within normal range.


Salvage chemotherapy with vinblastine, ifosfamide, and cisplatin in recurrent seminoma.
Salvage therapy for disseminated germ cell tumors of all histologic subtypes with vinblastine, ifosfamide, and cisplatin (VeIP) will cure approximately 25% of patients. The purpose of this study was to evaluate the activity of VeIP in patients with recurrent seminoma. We conducted a retrospective review of 24 patients with recurrent seminoma who were treated at Indiana University with VeIP as second-line chemotherapy. All patients had received cisplatin-containing regimens as primary chemotherapy and seven had also received prior pelvic radiotherapy. All patients received four courses of VeIP. The minimum follow-up duration was 2 years (range, 2 to 9.1), with a median follow-up time of 7 years. Twenty of 24 patients (83%) achieved a complete remission (CR) following VeIP alone. One additional patient was rendered disease-free (NED) with the resection of residual carcinoma. Eight patients have relapsed. Four of six patients with extragonadal primary tumors and two of four who failed to achieve CR with initial chemotherapy are continuously NED with VeIP. Overall, 13 of 24 (54%) are long-term survivors with VeIP salvage chemotherapy. VeIP has significant curative potential in patients with recurrent seminoma and appears to produce a higher CR rate and more long-term survivors than is achieved in patients with nonseminomatous disease.


Cyclopentane rings in hydrophobic chains of a phospholipid enhance the bilayer stability to electric breakdown.
Archaeal lipids ensure unprecedented stability of archaea membranes in extreme environments. Here, we incorporate a characteristic structural feature of an archaeal lipid, the cyclopentane ring, into hydrocarbon chains of a short-chain (C12) phosphatidylcholine to explore whether the insertion would allow such a lipid (1,2-di-(3-(3-hexylcyclopentyl)-propanoate)-sn-glycero-3-phosphatidylcholine, diC12cp-PC) to form stable bilayers at room temperature. According to fluorescence-based assays, in water diC12cp-PC formed liquid-crystalline bilayers at room temperature. Liposomes produced from diC12cp-PC retained calcein for over a week when stored at +4 °C. diC12cp-PC could also form model bilayer lipid membranes that were by an order of magnitude more stable to electrical breakdown than egg PC membranes. Molecular dynamics simulation showed that the cyclopentane fragment fixes five carbon atoms (or four C-C bonds), which is compensated by the higher mobility of the rest of the chain. This was found to be the reason for the remarkable stability of the diC12cp-PC bilayer: restricted conformational mobility of a chain segment increases the membrane bending modulus (compared to a normal hydrocarbon chain of the same length). Here, higher stiffness practically does not affect the line tension of a membrane pore edge. Rather it makes it more difficult for diC12cp-PC to rearrange in order to line the edge of a hydrophilic pore; therefore, fewer pores are formed.


Untargeted metabolomics of perfusate and their association with hypothermic machine perfusion and allograft failure.
Although hypothermic machine perfusion (HMP) is associated with improved kidney graft viability and function, the underlying biological mechanisms are unknown. Untargeted metabolomic profiling may identify potential metabolites and pathways that can help assess allograft viability and contribute to organ preservation. Therefore, in this multicenter study, we measured all detectable metabolites in perfusate collected at the beginning and end of deceased-donor kidney perfusion and evaluated their associations with graft failure. In our cohort of 190 kidney transplants, 33 (17%) had death-censored graft failure over a median follow-up of 5.0 years (IQR 3.0-6.1 years). We identified 553 known metabolites in perfusate and characterized their experimental and biological consistency through duplicate samples and unsupervised clustering. After perfusion-time adjustment and false discovery correction, six metabolites in post-HMP perfusate were significantly associated with death-censored graft failure, including alpha-ketoglutarate, 3-carboxy-4-methyl-5-propyl-2-furanpropanoate, 1-carboxyethylphenylalanine, and three glycerol-phosphatidylcholines. All six metabolites were associated with an increased risk of graft failure (Hazard Ratio per median absolute deviation range 1.04-1.45). Four of six metabolites also demonstrated significant interaction with donation after cardiac death with notably greater risk in the donation after cardiac death group (Hazard Ratios up to 1.69). Discarded kidneys did not have significantly different levels of any death-censored graft failure-associated metabolites. On interrogation of pathway analysis, production of reactive oxygen species and increased metabolism of fatty acids were upregulated in kidneys that subsequently developed death-censored graft failure. Thus, further understanding the role of these metabolites may inform the HMP process and help improve the objective evaluation of allograft offers, thereby reducing the discard of potentially viable organs.


Procalcitonin to initiate or discontinue antibiotics in acute respiratory tract infections.
Acute respiratory infections (ARIs) comprise of a large and heterogeneous group of infections including bacterial, viral, and other aetiologies. In recent years, procalcitonin (PCT), a blood marker for bacterial infections, has emerged as a promising tool to improve decisions about antibiotic therapy (PCT-guided antibiotic therapy). Several randomised controlled trials (RCTs) have demonstrated the feasibility of using procalcitonin for starting and stopping antibiotics in different patient populations with ARIs and different settings ranging from primary care settings to emergency departments, hospital wards, and intensive care units. However, the effect of using procalcitonin on clinical outcomes is unclear. This is an update of a Cochrane review and individual participant data meta-analysis first published in 2012 designed to look at the safety of PCT-guided antibiotic stewardship. The aim of this systematic review based on individual participant data was to assess the safety and efficacy of using procalcitonin for starting or stopping antibiotics over a large range of patients with varying severity of ARIs and from different clinical settings. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), which contains the Cochrane Acute Respiratory Infections Group's Specialised Register, MEDLINE, and Embase, in February 2017, to identify suitable trials. We also searched ClinicalTrials.gov to identify ongoing trials in April 2017. We included RCTs of adult participants with ARIs who received an antibiotic treatment either based on a procalcitonin algorithm (PCT-guided antibiotic stewardship algorithm) or usual care. We excluded trials if they focused exclusively on children or used procalcitonin for a purpose other than to guide initiation and duration of antibiotic treatment. Two teams of review authors independently evaluated the methodology and extracted data from primary studies. The primary endpoints were all-cause mortality and treatment failure at 30 days, for which definitions were harmonised among trials. Secondary endpoints were antibiotic use, antibiotic-related side effects, and length of hospital stay. We calculated odds ratios (ORs) and 95% confidence intervals (CIs) using multivariable hierarchical logistic regression adjusted for age, gender, and clinical diagnosis using a fixed-effect model. The different trials were added as random-effects into the model. We conducted sensitivity analyses stratified by clinical setting and type of ARI. We also performed an aggregate data meta-analysis. From 32 eligible RCTs including 18 new trials for this 2017 update, we obtained individual participant data from 26 trials including 6708 participants, which we included in the main individual participant data meta-analysis. We did not obtain individual participant data for four trials, and two trials did not include people with confirmed ARIs. According to GRADE, the quality of the evidence was high for the outcomes mortality and antibiotic exposure, and quality was moderate for the outcomes treatment failure and antibiotic-related side effects.Primary endpoints: there were 286 deaths in 3336 procalcitonin-guided participants (8.6%) compared to 336 in 3372 controls (10.0%), resulting in a significantly lower mortality associated with procalcitonin-guided therapy (adjusted OR 0.83, 95% CI 0.70 to 0.99, P = 0.037). We could not estimate mortality in primary care trials because only one death was reported in a control group participant. Treatment failure was not significantly lower in procalcitonin-guided participants (23.0% versus 24.9% in the control group, adjusted OR 0.90, 95% CI 0.80 to 1.01, P = 0.068). Results were similar among subgroups by clinical setting and type of respiratory infection, with no evidence for effect modification (P for interaction > 0.05). Secondary endpoints: procalcitonin guidance was associated with a 2.4-day reduction in antibiotic exposure (5.7 versus 8.1 days, 95% CI -2.71 to -2.15, P < 0.001) and lower risk of antibiotic-related side effects (16.3% versus 22.1%, adjusted OR 0.68, 95% CI 0.57 to 0.82, P < 0.001). Length of hospital stay and intensive care unit stay were similar in both groups. A sensitivity aggregate-data analysis based on all 32 eligible trials showed similar results. This updated meta-analysis of individual participant data from 12 countries shows that the use of procalcitonin to guide initiation and duration of antibiotic treatment results in lower risks of mortality, lower antibiotic consumption, and lower risk for antibiotic-related side effects. Results were similar for different clinical settings and types of ARIs, thus supporting the use of procalcitonin in the context of antibiotic stewardship in people with ARIs. Future high-quality research is needed to confirm the results in immunosuppressed patients and patients with non-respiratory infections.


Axonal damage revealed by accumulation of beta-APP in HIV-positive individuals without AIDS.
The presence of neuropsychological disturbances in HIV-positive, pre-symptomatic individuals is a controversial issue. Neuroimaging studies have not shown brain atrophy or hyperintensity in the white matter, whereas proton magnetic resonance spectroscopy has revealed some abnormality of cerebral biochemistry. Using an antibody to beta-amyloid precursor protein (beta-APP), we previously demonstrated frequent and widespread axonal changes in the brains of AIDS patients. In this study, we extended the use of beta-APP to asymptomatic patients in order to establish a possible morphological correlation with neuropsychological disorders. Brain samples from 29 patients were examined. Results showed bundles of beta-APP-positive axons in 8/29 cases (27%). The changes, seen in both superficial and deep white matter, were either focal or diffuse, could not be visualized by silver or ubiquitin stains, and did not coexist with any change in distribution or morphology of astrocytes and microglial cells. We conclude that in HIV-positive asymptomatic individuals, axonal changes: (a) may be related to the state of immune activation with consequent presence of toxic substances, including cytokines, observed in these patients; (b) may represent mild changes that could undergo repair, unless other pathological events, such as the supervening of the AIDS stage and the specific encephalitis, make them permanent.


Apramycin and gentamicin resistance in Escherichia coli and salmonellas isolated from farm animals.
Since the aminoglycoside antibiotic apramycin was licensed for veterinary use in 1980, all isolates of Escherichia coli and salmonellas received at the Central Veterinary Laboratory have been monitored for resistance to apramycin and the related antibiotic gentamicin. During the period 1982-4, the incidence of resistance in E. coli to apramycin increased from 0.6% in 1982 to 2.6% in 1984. In salmonellas the incidence of resistance to apramycin increased from 0.1% in 1982 to 1.4% in 1984. Resistance to both apramycin and gentamicin was detected in six different salmonella serotypes, although an isolate of Salmonella thompson from poultry was resistant to gentamicin but not apramycin. Most of the cultures were isolated from pigs, although the incidence of apramycin resistance in S. typhimurium (DT 204C) from calves has shown a recent dramatic increase. All the isolates with one exception produced the enzyme aminoglycoside 3-N-acetyltransferase IV (ACC(3)IV). The resistance was transferable by conjugation in most of the strains examined, and the plasmids specifying the resistance have been found to belong to a number of different incompatibility groups. Plasmids from three E. coli strains were compatible with all the reference plasmids and belonged to a previously undescribed group which was investigated further. It is suggested that bacteria from humans should be examined for resistance to apramycin and gentamicin to determine the possibility of the antibiotic-resistance bacteria, and their genes, spreading from animals to humans.


Delta 9-tetrahydrocannabinol inhibits arachidonic acid acylation of phospholipids and triacylglycerols in guinea pig cerebral cortex slices.
We reported earlier that delta 9-tetrahydrocannabinol (THC), the main psychoactive ingredient in marihuana, increased markedly the level of unesterified arachidonic acid (AA) in guinea pig cerebral cortex slices prelabeled with [14C]AA. The purpose of the present study was to clarify the mechanism underlying THC-enhanced mobilization of AA. We could find little data to support an involvement for phospholipase A2 in this response. For example, the levels of lysophosphatidylcholine or lysophosphatidylethanolamine were not elevated after incubation with THC. A role for phosphoinositidase C-initiated lipolytic pathways was excluded, because neither basal nor acetylcholine-stimulated inositol phosphate formation was altered by THC. When we prelabeled slices with [14C]stearate or [3H]glycerol, THC did not elevate levels of unesterified [14C]stearate, nor did we observe significant changes in the phospholipids that were labeled with either precursor. These findings were in marked contrast to the previously reported reductions in [14C]AA-labeled phosphatidylinositol after exposure of prelabeled brain slices to THC; moreover, they suggested that the THC-induced effects on brain lipid metabolism in vitro were rather specific for AA. We show here that, when the acylation of brain lipids with AA was measured by addition of [3H]AA in the presence and absence of THC at zero time and incubation for 1 hr at 37 degrees, THC elicited marked, dose-dependent, and saturable reductions in esterified [3H]AA levels. The reductions in incorporation were balanced by increases in unesterified [3H]AA. The IC50 for the effect was on the order of 8 microM, and a maximal response occurred at 32 microM. We observed that the THC-induced suppression in acylation of the phospholipids by radiolabeled AA was up to 5-fold greater than the THC-elicited loss of AA from slices prelabeled before exposure to THC. The largest inhibitions of acylation were in phosphatidylinositol; the suppression of radioactivity in this phospholipid accounted for over 50% of the rise in unesterified [3H]AA. The radioactivity incorporated in triacylglycerols were also reduced markedly by THC. In contrast, the incorporation of radioactivity in phosphatidylcholine remained unaffected by THC. Taken together, these findings suggest that THC mobilizes AA by inhibiting acylation of certain lipids with AA, particularly phosphatidylinositol and triacylglycerol, rather than by liberating fatty acids by lipolysis. Comparison of the effects of several primary cannabinoids on lipid acylation with [3H]AA revealed that there was no relationship between the potencies of cannabinoids in inhibiting the incorporation of [3H]AA into membrane lipids and their psychoactive potencies in vivo; moreover, the stereoisomers of THC were equipotent.(ABSTRACT TRUNCATED AT 400 WORDS)


Personality, Motivation, and Math Achievement Among Turkish Students.
Using the Turkish portion of the Programme for International Student Assessment dataset ( N = 4,848; 51% boys, 49% girls; age, M = 15.81 years, SD = 0.28), this study investigated factors associated with mathematics achievement among Turkish students. Three different models were estimated using the method of balanced repeated replication with Fay's method and taking into account the presence of five plausible values of the dependent variable. Results showed that male students and older students had better mathematics proficiency. Socio-economic status and school resources also played a significant role in explaining student achievement in mathematics. Finally, students who were more open to problem solving, who attributed their failure to external factors, and who were intrinsically motivated to learn mathematics achieved higher scores. Policy implications are provided.


COPD and the risk of tuberculosis--a population-based cohort study.
Both chronic obstructive pulmonary disease (COPD) and tuberculosis (TB) primarily affect the lungs and are major causes of morbidity and mortality worldwide. COPD and TB have common risk factors such as smoking, low socioeconomic status and dysregulation of host defence functions. COPD is a prevalent co-morbid condition, especially in elderly with TB but in contrast to other diseases known to increase the risk of TB, relatively little is known about the specific relationship and impact from COPD on TB-incidence and mortality. All individuals > or = 40 years of age, discharged with a diagnosis of COPD from Swedish hospitals 1987-2003 were identified in the Swedish Inpatient Register (n = 115,867). Records were linked to the Swedish Tuberculosis Register 1989-2007 and the relative risk of active TB in patients with COPD compared to control subjects randomly selected from the general population (matched for sex, year of birth and county of residence) was estimated using Cox regression. The analyses were stratified by year of birth, sex and county of residence and adjusted for immigration status, socioeconomic status (SES) and inpatient co-morbidities previously known to increase the risk of TB. COPD patients had a three-fold increased hazard ratio (HR) of developing active TB (HR 3.0 (95% confidence interval 2.4 to 4.0)) that was mainly dependent on an increased risk of pulmonary TB. In addition, logistic regression estimates showed that COPD patients who developed active TB had a two-fold increased risk of death from all causes within first year after the TB diagnosis compared to the general population control subjects with TB (OR 2.2, 95% confidence interval 1.2 to 4.1). This population-based study comprised of a large number of COPD patients shows that these patients have an increased risk of developing active TB compared to the general population. The results raise concerns that the increasing global burden of COPD will increase the incidence of active TB. The underlying contributory factors need to be disentangled in further studies.


Effector-triggered post-translational modifications and their role in suppression of plant immunity.
Plant-pathogen interactions feature complex signaling exchanges between host and microbes that ultimately determine association outcomes. Plants deploy pattern recognition receptors to perceive pathogen-associated molecular patterns, mount pattern-triggered immunity (PTI), and fend off potential pathogens. In recent years an increasing number of defense-signaling components have been identified along with a mechanistic understanding of their regulation during immune responses. Post-translational modifications (PTMs) are now thought to play a crucial role in regulating defense signaling. In a bid to suppress PTI and infect their host, pathogens have evolved large repertoires of effectors that trigger susceptibility and allow colonization of host tissues. While great progress has been made in elucidating defense-signaling networks in plants and the activities of effectors in immune suppression, a critical gap exists in our understanding of effector mechanism-of-action. Given the importance of PTMs in the regulation of defense signaling, we will explore the question: how do effectors modify the post-translational status of host proteins and thus interfere with host processes required for immunity? We will consider how emerging proteomics-based experimental strategies may help us answer this important question and ultimately open the pathogens' effector black box.


Can individual fatty acids be used as functional biomarkers of dairy fat consumption in relation to cardiometabolic health? A narrative review.
In epidemiological studies, dairy food consumption has been associated with minimal effect or decreased risk of some cardiometabolic diseases (CMD). However, current methods of dietary assessment do not provide objective and accurate measures of food intakes. Thus, the identification of valid and reliable biomarkers of dairy product intake is an important challenge to best determine the relationship between dairy consumption and health status. This review investigated potential biomarkers of dairy fat consumption, such as odd-chain, trans- and branched-chain fatty acids (FA), which may improve the assessment of full-fat dairy product consumption. Overall, the current use of serum/plasma FA as biomarkers of dairy fat consumption is mostly based on observational evidence, with a lack of well-controlled, dose-response intervention studies to accurately assess the strength of the relationship. Circulating odd-chain SFA and trans-palmitoleic acid are increasingly studied in relation to CMD risk and seem to be consistently associated with a reduced risk of type 2 diabetes in prospective cohort studies. However, associations with CVD are less clear. Overall, adding less studied FA such as vaccenic and phytanic acids to the current available evidence may provide a more complete assessment of dairy fat intake and minimise potential confounding from endogenous synthesis. Finally, the current evidence base on the direct effect of dairy fatty acids on established biomarkers of CMD risk (e.g. fasting lipid profiles and markers of glycaemic control) mostly derives from cross-sectional, animal and in vitro studies and should be strengthened by well-controlled human intervention studies.


Microtubule-associated protein 1B dysregulates microtubule dynamics and neuronal mitochondrial transport in spinal muscular atrophy.
Spinal muscular atrophy (SMA) is a devastating childhood disease primarily affecting lower motoneurons in the spinal cord. SMA is caused by the loss of functional survival of motoneuron (SMN) protein, leading to structural and functional alterations of the cytoskeleton in motoneurons and other cells. Loss of SMN results in impairments of microtubule architecture, but the underlying mechanisms are not completely understood. In this study, we mechanistically analyzed the effects of SMN deficiency on microtubules, demonstrating a reduced stability together with a reduction in alpha tubulin detyrosination. This was caused by increased levels of microtubule-associated protein 1B and tubulin tyrosine ligase, resulting in mitochondrial mislocalization in SMA. Our findings suggest that altered tubulin post-translational modifications and microtubule-associated proteins are involved in the pathomechanisms of SMA, such as an impaired axonal transport of mitochondria.


Prediction of Critical Buckling Load on Open Cross-Section Columns of Flax/PLA Green Composites.
The present work aims to analyze the buckling behavior of nonlinear elastic columns with different open cross-sections and slenderness ratios to verify the limits of the modified Ludwick law to predict the critical buckling load. The results of the analytical formulation based on the modified Ludwick law are compared with a FEM numerical model using the Marlow hyperelastic behavior and experimental results conducted on flax/PLA specimens with three different open cross-sections. The comparative results show that the numerical predictions agree with the experimental results in all the cases. The FEM model can exactly reproduce the buckling behavior of the C-section columns. However, the prediction errors for the C90 and C180 columns are higher than for the C60 columns. Moreover, the theoretical estimations indicate that the C90 cross-section column is the limit of application of the modified Ludwick law to predict the critical buckling load of nonlinear elastic columns with open cross-sections, and the C180 column is out of the prediction limits. Generally, the numerical and theoretical models underestimated the scattering effects of the predictions because more experimental variables were not considered by the models.


Association between serum uric acid and homocysteine levels among adults in the United States: a cross-sectional study.
Many studies have shown that both elevated serum uric acid (SUA) levels and hyperhomocysteinemia are risk factors for atherosclerosis. However, the relationship between the two has not been thoroughly investigated. This study aimed to explore the possible link between SUA levels and homocysteine (Hcy) levels. In this cross-sectional study, 17,692 adults aged > 19 years in National Health and Nutrition Examination Survey from 1999 to 2006 were analyzed. Multivariable linear regression analysis was performed to assess the association between SUA and Hcy levels. In addition, smooth curve fitting (penalized spline method) and threshold effect analysis were performed. Multivariable linear analysis showed that Hcy levels increased by 0.48 µmol/L (β = 0.48, 95%CI: 0.43-0.53) for every 1 mg/dL increase in SUA levels. We found a nonlinear relationship between SUA and Hcy levels. The results of threshold effect analysis showed that the inflection point for SUA levels was 7.1 mg/dL (β = 0.29, 95% CI: 0.23-0.36 and β = 1.05, 95% CI: 0.67-1.43 on the left and right sides of the inflection point, respectively). The p-values was less than 0.001 when using the log likelihood ratio test. This nonlinear relationship was also found in both sexes. The inflection point for SUA levels was 5.4 mg/dL in males and 7.3 mg/dL in females, respectively. This cross-sectional study showed that the SUA levels were positively correlated with Hcy levels. And we found a nonlinear relationship between SUA and Hcy levels.


HLA-DQ genotype is associated with accelerated small bowel transit in patients with diarrhea-predominant irritable bowel syndrome.
Colonic transit (CT) is accelerated in 46% of patients with diarrhea-predominant irritable bowel syndrome (IBS-D). Improvement in IBS-D with gluten withdrawal is associated with human leukocyte antigen (HLA)-DQ2 positivity; the mechanism of improvement is unclear. To determine if HLA-DQ2-positive or HLA-DQ8-positive patients with IBS-D have faster small bowel (SB) or CT than HLA-DQ2-negative and HLA-DQ8-negative patients. Among 94 patients with IBS-D, who previously provided DNA samples, 64 had undergone validated measurements of CT [geometric center at 24 h (GC24)]; 50 of the patients also had measurement of gastric emptying (GE) and 54 of SB transit (colonic filling at 6 h). HLA-DQ status was determined by tag single nucleotide polymorphism approach. Associations of colonic filling at 6 h and GC24 with HLA-DQ2 and HLA-DQ8 status were assessed using analysis of covariance, adjusting for BMI. Mean age was 40.8±1.6 years; 98.5% were females. In 60 of the 64 patients, celiac disease was excluded by serology or histology. There were no significant differences in age or BMI among the different HLA-DQ groups. Independently, patients positive for HLA-DQ2 had numerically greater colonic filling at 6 h compared with HLA-DQ2-negative (P=0.065), and those positive for HLA-DQ8 had greater colonic filling at 6 h compared with HLA-DQ8-negative patients (P=0.021). Gastric emptying was not associated with HLA-DQ2 and HLA-DQ8 status. Patients positive for both HLA-DQ2 and HLA-DQ8 had greater colonic filling at 6 h (P=0.013) and numerically higher, but not significant, GC24 (P=0.38) compared with HLA-DQ2-negative and HLA-DQ8-negative patients. Patients with IBS-D positive for HLA-DQ8 or for both HLA-DQ2 and HLA-DQ8 have faster SB transit. The mechanism of the accelerated SB transit and the effect of gluten withdrawal on SB function in IBS-D deserve further investigation.


Quantifying the human impact of Melbourne's 111-day hard lockdown experiment on the adult population.
Lockdown was used worldwide to mitigate the spread of severe acute respiratory syndrome coronavirus 2 and was the cornerstone non-pharmaceutical intervention of zero-COVID strategies. Many previous impact evaluations of lockdowns are unreliable because lockdowns co-occurred with severe coronavirus disease related health and financial insecurities. This was not the case in Melbourne's 111-day lockdown, which left other Australian jurisdictions unaffected. Interrogating nationally representative longitudinal survey data and quasi-experimental variation, and controlling for multiple hypothesis testing, we found that lockdown had some statistically significant, albeit small, impacts on several domains of human life. Women had lower mental health (-0.10 s.d., P = 0.043, 95% confidence interval (CI) = -0.21 to -0) and working hours (-0.13 s.d., P = 0.006, 95% CI = -0.22 to -0.04) but exercised more often (0.28 s.d., P < 0.001, 95% CI = 0.18 to 0.39) and received more government transfers (0.12 s.d., P = 0.048, 95% CI = 0.001 to 0.24). Men felt less part of their community (-0.20 s.d., P < 0.001, 95% CI = -0.30 to -0.10) and reduced working hours (-0.12 s.d., P = 0.004, 95% CI = -0.20 to -0.04). Heterogeneity analyses demonstrated that families with children were driving the negative results. Mothers had lower mental health (-0.27 s.d., P = 0.014, 95% CI = -0.48 to -0.06), despite feeling safer (0.26 s.d., P = 0.008, 95% CI = 0.07 to 0.46). Fathers increased their alcohol consumption (0.35 s.d., P = 0.002, 95% CI = 0.13 to 0.57). Some outcomes worsened with lockdown length for mothers. We discuss potential explanations for why parents were adversely affected by lockdown.


Effects of Age and Gender on Hand Motion Tasks.
Objective. Wearable and wireless motion sensor devices have facilitated the automated computation of speed, amplitude, and rhythm of hand motion tasks. The aim of this study is to determine if there are any biological influences on these kinematic parameters. Methods. 80 healthy subjects performed hand motion tasks twice for each hand, with movements measured using a wireless motion sensor device (Kinesia, Cleveland Medical Devices Inc., Cleveland, OH). Multivariate analyses were performed with age, gender, and height added into the model. Results. Older subjects performed poorer in finger tapping (FT) speed (r = 0.593, p < 0.001), hand-grasp (HG) speed (r = 0.517, p < 0.001), and pronation-supination (PS) speed (r = 0.485, p < 0.001). Men performed better in FT rhythm (p < 0.02), HG speed (p < 0.02), HG amplitude (p < 0.02), and HG rhythm (p < 0.05). Taller subjects performed better in the speed and amplitude components of FT (p < 0.02) and HG tasks (p < 0.02). After multivariate analyses, only age and gender emerged as significant independent factors influencing the speed but not the amplitude and rhythm components of hand motion tasks. Gender exerted an independent influence only on HG speed, with better performance in men (p < 0.05). Conclusions. Age, gender, and height are not independent factors influencing the amplitude and rhythm components of hand motion tasks. The speed component is affected by age and gender differences.


HIV infection and its association with an excess risk of clinical fractures: a nationwide case-control study.
Different studies have reported an association between HIV infection, antiretroviral therapies, and impaired bone metabolism, but data on their impact on fracture risk are scarce. We studied the association between a clinical diagnosis of HIV infection and fracture risk. We conducted a case-control study using data from the Danish National Health Service registries, including 124,655 fracture cases and 373,962 age- and gender-matched controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using conditional logistic regression. A total of 50 (0.40/1000) patients in the fracture group and 52 (0.14/1000) controls had an HIV diagnosis. The risk of any fracture was thus significantly increased among HIV-infected patients (age- and gender-matched OR = 2.89, 95% CI: 1.99 to 4.18). Similarly, significant increases in the risk of hip (OR = 8.99, 95% CI: 1.39 to 58.0), forearm (OR = 3.50, 95% CI: 1.26 to 9.72), and spine fractures (OR = 9.00, 95% CI: 1.39 to 58.1) were observed. HIV infection is associated with an almost 3-fold increase in fracture risk compared with that of age- and gender-matched uninfected patients. HIV patients are also at an almost 9-fold higher risk of hip fracture.


Neuronal NOS gene expression in gastrointestinal myenteric neurons and smooth muscle cells.
Nitric oxide synthase (NOS) has been characterized in different tissues, and its localization has been suggested in different neuronal tissues, including the myenteric neurons and other nonneuronal cells. The present study examined the distribution of the neuronal NOS (nNOS) mRNA in different tissues of the opossum gastrointestinal tract, internal anal sphincter (IAS) smooth muscle cells, and myenteric neurons using slot-blot and Northern blot hybridization techniques with a specific rat brain nNOS cDNA probe. Significant levels of nNOS gene expression were found in both smooth muscle cells and myenteric neurons of the opossum IAS. This finding was confirmed by reverse transcriptase-polymerase chain reaction analysis of the RNA obtained from cultured opossum IAS smooth muscle cells and myenteric neurons and also from human intestinal smooth muscle and neuroblastoma cell lines. Pyloric sphincter had the highest level of nNOS gene expression compared with other gastrointestinal tissues. There was no significant difference in the nNOS gene expression between other sphincteric and nonsphincteric tissues examined. The present study shows the presence of nNOS gene expression in both neurons and smooth muscle cells. The higher levels of nNOS gene expression in the pyloric sphincter compared with other tissues may have pathophysiological significance in some disease conditions.


[The structure, growth and formation of bones in toxic exposure of the body to pesticides and antioxidant therapy].
In the experiment performed on 175 white male rats by means of a complex of morphological, biochemical and biomechanical methods peculiarities of structure, growth, outline formation and mineralization of the skeletal bones have been investigated under a toxic lesion of the organism with pesticides (chlorophose and keltan) and at a simultaneous administration of antioxidants of various groups (tocopherol, ionol, sodium selenit). Osteotoxic effect of the pesticides, manifested as an inhibition of bone growth, as a disbalance of mineral saturation and their composition, as a decrease in indices of firmness is leveled by means of therapeutic doses of the antioxidants. The stabilizing effect of the antioxidant applied correlates to the manifestation of the pesticides osteotoxic effect, to the ability of their cumulation in the organism and is directly connected with the supposed mechanism of damaging effect to the organism and/or cell. The expressiveness of the toxic effect of the chemical poison, in its turn, is defined not only by the dose, mechanism and duration of the effect, but by age peculiarities of the organism and by functional state of its reactivity. When the poisons are applied for a long time, in order to level their osteotoxic effect, a multiple increase of therapeutic doses of the antioxidants and a combined potentiation of their effect are necessary.


Honey use in burn management: potentials and limitations.
Management of the burn wound still remains a matter of debate, and an ideal dressing for burn wounds has not yet been discovered. Naturally occurring substances such as honey have been found to be useful as a wound cover for burns. Unlike most conventional local chemotherapeutics, honey does not lead to the development of antibiotic-resistant bacteria, and it may be used continuously. Among the challenging problems of using honey for medical purposes are dosage, safety, and formulation. Many approaches have been suggested to overcome such problems. With the increased availability of licensed medical products containing honey, clinical use is expected to increase and further evidence will become available. Honey seems to have the potential to clear infection as well as to be an effective prophylactic agent that may contribute to reducing the risks of cross-infection. A better understanding of the therapeutic and chemical properties of honey is needed to optimise the use of this product in the clinical management of burns. Its use in professional care centres should be limited to those with certified healing activities. The potentials and limitations of using honey as burn dressing are discussed in this review.


The effect of a 980 nm diode laser with different parameters of irradiation on the bond strength of fiberglass posts.
The aim of this study was to assess bond strength of fiberglass posts to root canal dentin irradiated with a 980 nm diode laser at different parameters of power and frequency. Fifty human maxillary canines were separated into five groups (n = 10) according to the following parameters of laser power and frequency: Group 1 (1.5 W/100 Hz), Group 2 (1.5 W/continuous wave [CW]), Group 3 (3.0 W/100 Hz), Group 4 (3.0 W/CW), and Group 5 (no irradiation). Following post cementation, samples underwent a push-out test (0.5 mm/min); next, fracture analysis was performed with a light microscope at 50x and 100x magnification. All of the irradiated groups had increased bond strength values compared to the nonirradiated group. Groups 1 and 2 demonstrated the highest bond strength values; however, statistically significant differences were observed for only the cervical third of Group 4 and the cervical/apical thirds of Group 5. Fracture analysis showed a predominance of mixed failures for Groups 1 and 2 and adhesive failures between dentin and cement for the other groups.


Detection of a quantitative trait locus associated with resistance to infection with Trichuris suis in pigs.
Whipworms (Trichuris spp.) infect a variety of hosts, including domestic animals and humans. Of considerable interest is the porcine whipworm, T. suis, which is particularly prevalent in outdoor production systems. High infection levels may cause growth retardation, anaemia and haemorrhagic diarrhoea. A significant proportion of the variation in Trichuris faecal egg count (FEC) has been attributed to the host's genetic make-up. The aim of the present study was to identify genetic loci associated with resistance to T. suis in pigs. We used single nucleotide polymorphism (SNP) markers to perform a whole-genome scan of an F1 resource population (n = 195) trickle-infected with T. suis. A measured genotype analysis revealed a putative quantitative trait locus (QTL) for T. suis FEC on chromosome 13 covering ∼ 4.5 Mbp, although none of the SNPs reached genome-wide significance. We tested the hypothesis that this region of SSC13 harboured genes with effects on T. suis burden by genotyping three SNPs within the putative QTL in unrelated pigs exposed to either experimental or natural T. suis infections and from which we had FEC (n = 113) or worm counts (n = 178). In these studies, two of the SNPs (rs55618716, ST) were associated with FEC (P < 0.01), thus confirming our initial findings. However, we did not find any of the SNPs to be associated with T. suis worm burden. In conclusion, our study demonstrates that genetic markers for resistance to T. suis as indicated by low FEC can be identified in pigs.


Direct Proof of Endo-Epicardial Asynchrony of the Atrial Wall During Atrial Fibrillation in Humans.
The presence of focal fibrillation waves during atrial fibrillation (AF) can, besides ectopic activity, also be explained by asynchronous activation of the atrial endo- and epicardial layer and transmurally propagating fibrillation waves. To provide direct proof of endo-epicardial asynchrony, we performed simultaneous high-resolution mapping of the right atrial endo- and epicardial wall during AF in humans. Intraoperative mapping of the endo- and epicardial right atrial wall was performed during (induced) AF in 10 patients with AF (paroxysmal: n=3; persistent: n=4; and longstanding persistent: n=3) and 4 patients without a history of AF. A clamp made of 2 rectangular 8×16 electrode arrays (interelectrode distance 2 mm) was inserted into the incision in the right atrial appendage. Recordings of 10 seconds of AF were analyzed to determine the incidence of asynchronous endo-epicardial activation times (≥15 ms) of opposite electrodes. Asynchronous endo-epicardial activation ranged between 0.9 and 55.9% without preference for either side. Focal waves appeared equally frequent at endocardium and epicardium (11% versus 13%; ITALIC! P=0.18). Using strict criteria for breakthrough (presence of an opposite wave within 4 mm and ≤14 ms before the origin of the focal wave), the majority (65%) of all focal fibrillation waves could be attributed to endo-epicardial excitation. We provided the first evidence for asynchronous activation of the endo-epicardial wall during AF in humans. Endo-epicardial asynchrony may play a major role in the pathophysiology of AF and may offer an explanation why in some patients therapy fails.


A Kernel Reinforcement Learning Decoding Framework Integrating Neural and Feedback Signals for Brain Control.
Brain-Machine Interfaces (BMIs) have the potential to allow subjects to brain control (BC) external devices, where their brain signals could be translated to the action of the neuro-prosthesis by reinforcement learning (RL) based decoder. During the BC task, feedback cues are provided to guide subject's learning. Subjects will adapt the neural signals according to the feedback cues. Concurrently, the RL decoding parameters are adjusted when the subject explores the BC task through trial and error, leading to a co-adaptive process between the subject and the decoder. However, when subjects receive the feedback cues and enhance their learning, the decoder does not actively utilize the feedback cues. If the RL decoder could integrate both neural signals and feedback cues, the training efficiency of the BC task would increase. A major challenge is the different temporal scales of neural signals and feedback cues, making it difficult to integrate them into a single decoder. In this paper, we propose a novel kernel RL decoding method as the first attempt to combine two signals with different temporal scales for RL decoding. The neural signals and the feedback cues comprise the decoding input, which is then projected into individual Reproducing Kernel Hilbert Spaces (RKHSs) respectively. These two RKHSs form a joint feature space, where the action of the neuro-prosthesis could be decoded linearly. We evaluate the proposed method on a simulated brain control cursor-reaching task. Our proposed method is compared with the kernel RL that only uses neural signals as the input. The proposed method has a faster learning speed and better decoding accuracy. The results demonstrate that our proposed method has successfully integrated the information of the feedback cue and facilitates the training procedure for the BC task.Clinical Relevance-This paper provides an integrated reinforcement learning decoding framework, which combines the neural signals and the feedback cues to increase the learning speed and the accuracy of the brain control task. Subjects could learn the task more easily with this decoder.


Dual‑directional effect of vinorelbine combined with cisplatin or fluorouracil on tumor growth and metastasis in metronomic chemotherapy in breast cancer.
Metronomic chemotherapy (MCT) regimens may be associated with risks to the patient due to the ambiguity surrounding low dosages and schedules. In the present study, metronomic regimens of vinorelbine (NVB) combined with cisplatin (CDDP) or fluorouracil (5‑FU) were chosen to study the dose‑response associations with tumor growth and metastasis, along with the underlying mechanisms in angiogenesis, apoptosis and tumor immunity, using experimental techniques such as immunofluorescence, immunohistochemistry, western blotting and flow cytometry. The results demonstrated a dual‑directional pharmacological action of promoting and suppressing tumor growth or metastasis in BALB/c mice bearing a 4T1 tumor at certain low and high doses of the drugs. Low doses of NVB combined with CDDP or 5‑FU accelerated tumor growth by enhancing angiogenesis, increasing the expression of angiogenic proteins, NF‑κB and osteopontin in tumor tissues, and inducing the accumulation of myeloid‑derived suppressor cells and macrophages. By contrast, higher doses inhibited tumor growth by suppressing these effects. Notably, the upregulation of apoptotic proteins was observed after low‑ and high‑dose treatments. Furthermore, at low concentrations, NVB combined with CDDP or 5‑FU stimulated certain functions of endothelial and tumor cells, including migration and invasion, whereas at higher concentrations they suppressed proliferation and induced apoptosis. Therefore, the results of the present study suggested the potential risks of metronomic combination chemotherapy by demonstrating that, at certain low doses, tumor growth or metastasis was promoted, and emphasized the existence of an effective dose interval that changes with different drug combinations. However, further studies are needed before a specific metronomic combination regimen can be administered clinically for cancer treatment.


Interleukin 1 type 1 receptor restore: a genetic mouse model for studying interleukin 1 receptor-mediated effects in specific cell types.
Interleukin-1 (IL-1) mediates diverse neurophysiological and neuropathological effects in the CNS through type I IL-1 receptor (IL-1R1). However, identification of IL-1R1-expressing cell types and cell-type-specific functions of IL-1R1 remains challenging. In this study, we created a novel genetic mouse model in which IL-1R1 gene expression is disrupted by an intronic insertion of a loxP flanked disruptive sequence that can be deleted by Cre recombinase, resulting in restored IL-1R1 gene expression under its endogenous promoters. A second mutation was introduced at stop codon of the IL-1R1 gene to allow tracking of the restored IL-1R1 protein by a 3HA tag and IL-1R1 mRNA by tdTomato fluorescence. These animals were designated as IL-1R1(r/r) and exhibited an IL-1R1 knock-out phenotype. We used IL-1R1 globally restored mice (IL-1R1(GR/GR)) as an IL-1R1 reporter and observed concordant labeling of IL-1R1 mRNA and protein in brain endothelial cells. Two cell-type-specific IL-1R1 restore lines were generated: Tie2Cre-IL-1R1(r/r) and LysMCre-IL-1R1(r/r). Brain endothelial COX-2 expression, CNS leukocyte infiltration, and global microglia activation induced by intracerebroventricular injection of IL-1β were not observed in IL-1R1(r/r) or LysMCre-IL-1R1(r/r) mice, but were restored in Tie2Cre-IL-1R1(r/r) mice. These results reveal IL-1R1 expression in endothelial cells alone is sufficient to mediate these central IL-1-induced responses. In addition, ex vivo IL-1β stimulation increased IL-1β expression in bone marrow cells in wild-type, Tie2Cre-IL-1R1(r/r), and LysMCre-IL-1R1(r/r), but not IL-1R1(r/r) mice. These results demonstrate this IL-1R1 restore model is a valuable tool for studying cell-type-specific functions of IL-1R1.


MULTIPLE EPIDEMICS IN AUSTRIAN FRINGILLIDAE CAUSED BY A SINGLE VARIANT OF SALMONELLA TYPHIMURIUM.
In Austria, numerous deaths of wild birds of the order Passeriformes, family Fringillidae, occurred during the winter months of 2010 and 2012. The Eurasian Siskin (Carduelis spinus) was the species most often affected. The dead birds were mainly found in the immediate vicinity of feeding places. Vigilant citizens sent birds (n=34) for pathologic examination to the Institute of Pathology in Vienna, Austria. All birds were cachectic or in a very poor nutritional condition. At gross examination, the most striking findings were multifocal to confluent, yellow-white nodules in the crop or esophageal mucosa. In histologically examined birds (n=24), severe transmural fibrino-purulent to necrotizing ingluviitis or esophagitis with large amounts of intralesional bacteria was observed. Bacteriologic examination of crop, liver, or other organs from 14 birds revealed abundant growth of Salmonella serovar Typhimurium (antigenic formula 1,4,5,12:i:1,2; phage type U277) in all individuals. By means of immunohistochemistry, these bacteria were detected not only in crop mucosa but also in lung tissue. In 17 birds (71%), structures morphologically resembling Macrorhabdus ornithogaster were detected histologically on the surface of the proventricular epithelium. Thus, the cause of mass mortality of the passerine birds was identified as infection with Salmonella Typhimurium, which was associated with growth of presumed M. ornithogaster in many cases.


Attention deficits in childhood-onset schizophrenia: reaction time studies.
The hypothesis of continuity between childhood-onset and adult schizophrenia was tested by comparing the performance of 15 patients with childhood-onset schizophrenia and 52 age-matched controls on 2 reaction time paradigms that have been used to study adult schizophrenia. On simple reaction time to tones with regular and irregular preparatory intervals of 2, 4, and 8 s, patients showed greater effects of the length of the preparatory interval in the regular condition and greater effects of the preparatory interval (girls only) and the preceding preparatory interval in the irregular series. On simple reaction time to random lights and tones, patients were faster on ipsimodal sequences than cross-modal sequences compared with controls. Overall, patients were much slower than controls in both paradigms. The results suggest similar attention dysfunction as is found in adult schizophrenia and thus are consistent with the continuity hypothesis.


Surface molecular imprints of WGA lectin as artificial receptors for mass-sensitive binding studies.
Wheat germ agglutinin (WGA) lectin is a model compound for the interaction between viruses and cells during infection events and thus an interesting analyte for mass-sensitive sensing to study these interaction phenomena. Scanning tunneling microscopy studies reveal that surface molecular imprinting leads to cavities having the dimensions of WGA dimers. These reincorporate WGA from phosphate-buffered saline between 1 and 160 μg/ml. Whereas the quartz crystal microbalance (QCM) frequency for molecularly imprinted polymer (MIP)-coated electrodes decreases, indicating uptake of the analyte, their nonimprinted counterparts yield positive, concentration-dependent frequency shifts characteristic for slip of the analyte on the QCM surface. The MIPs achieve selectivity factors towards bovine serum albumin of roughly 4 at higher protein concentrations. Brunauer-Emmett-Teller analysis reveals that binding is favored by 29 kJ/mol until the adsorption of up to ten monolayers on the MIP, whereas above this range the value is lower. Together with the binding behavior of MIP and nonimprinted polymers, this indicates that the MIP acts as a nucleus for multilayer deposition onto the surface.


Effect of APOE Genotype on Plasma Docosahexaenoic Acid (DHA), Eicosapentaenoic Acid, Arachidonic Acid, and Hippocampal Volume in the Alzheimer's Disease Cooperative Study-Sponsored DHA Clinical Trial.
Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (AA) play key roles in several metabolic processes relevant to Alzheimer's disease (AD) pathogenesis and neuroinflammation. Carrying the APOEɛ4 allele (APOE4) accelerates omega-3 polyunsaturated fatty acid (PUFA) oxidation. In a pre-planned subgroup analysis of the Alzheimer's Disease Cooperative Study-sponsored DHA clinical trial, APOE4 carriers with mild probable AD had no improvements in cognitive outcomes compared to placebo, while APOE 4 non-carriers showed a benefit from DHA supplementation. We sought to clarify the effect of APOEɛ4/ɛ4 on both the ratio of plasma DHA and EPA to AA, and on hippocampal volumes after DHA supplementation. Plasma fatty acids and APOE genotype were obtained in 275 participants randomized to 18 months of DHA supplementation or placebo. A subset of these participants completed brain MRI imaging (n = 86) and lumbar punctures (n = 53). After the intervention, DHA-treated APOEɛ3/ɛ3 and APOEɛ2/ɛ3 carriers demonstrated significantly greater increase in plasma DHA/AA compared to ɛ4/ɛ4 carriers. APOEɛ2/ɛ3 had a greater increase in plasma EPA/AA and less decline in left and right hippocampal volumes compared to compared to ɛ4/ɛ4 carriers. The change in plasma and cerebrospinal fluid DHA/AA was strongly correlated. Greater baseline and increase in plasma EPA/AA was associated with a lower decrease in the right hippocampal volume, but only in APOE 4 non-carriers. The lower increase in plasma DHA/AA and EPA/AA in APOEɛ4/ɛ4 carriers after DHA supplementation reduces brain delivery and affects the efficacy of DHA supplementation.


Clinical Outcome of Immediate Transurethral Surgery for Benign Prostate Obstruction Patients with Acute Urinary Retention: More Radical Resection Resulted in Better Voiding Function.
Transurethral surgery of the prostate is currently the gold standard treatment modality for patients with benign prostatic hyperplasia (BPH) with recurrent acute urinary retention. This study aimed to evaluate the outcome and predictors of patients receiving immediate surgery after acute urinary retention (AUR) episodes. From January 2016 to January 2017, we retrospectively included 714 patients who received transurethral surgery of prostate due to BPH. Among them, 158 patients received surgeries immediately after an AUR episode. General characteristics data including age, Body mass index (BMI), International prostate symptom score (IPSS score), prostate volume and Prostate-specific antigen (PSA) were reviewed. We also collected surgery-related parameters including surgical types, operation time, and specimen weight. Resection ratio was defined as (resected specimen weight)/(Transurethral ultrasound (TRUS) volume). The catheterization status on discharge, post-operative medication for BPH, and AUR within 3 months after operation were evaluated. Statistical analysis was performed with Statistical product and service solutions (SPSS). The mean age of the patients was 73.5 years, with a BMI of 24 kg/m2. IPSS total score was 25 with a Voiding symptom/Storage symptom score (V/S score) of 14.6 and 10.4, respectively. A total of 74 (46.8%) patients still took medication for BPH for over 1 month after the surgeries, 28 (17.7%) patients were not catheter-free at the time of discharge, and 14 (8.9%) patients had AUR within 3 months after the surgeries. Surgical type did not impact the outcome of surgeries. In patients who received Transurethral resection of the prostate (TURP), the resection ratio is the only predictor for the catheterization status on discharge and post-operative medication for BPH within 3 months after operation. In patients with BPH-related AUR followed by immediate transurethral surgeries, more radical resection is significantly correlated with short-term medication-free and catheter-free status.


Development of methane conversion factor models for Zebu beef cattle fed low-quality crop residues and by-products in tropical regions.
The enteric methane conversion factor (Ym) is an important country-specific value for the provision of precise enteric methane emissions inventory reports. The objectives of this meta-analysis were to develop and evaluate the empirical Ym models for the national level and the farm level for tropical developing countries according to the IPCC's categorization. We used datasets derived from 18 in vivo feeding experiments from 1999 to 2015 of Zebu beef cattle breeds fed low-quality crop residues and by-products. We found that the observed Ym value was 8.2% gross energy (GE) intake (~120 g methane emission head-1 day-1) and ranged from 4.8% to 13.7% GE intake. The IPCC default model (tier 2, Ym = 6.5% ± 1.0% GE intake) underestimated the Ym values by up to 26.1% compared with its refinement of 8.4% ± 0.4% GE intake for the national-level estimate. Both the IPCC default model and the refined model performed worse in predicting Ym trends at the farm level (root mean square prediction error [MSPE] = 15.1%-23.1%, concordance correlation coefficient [CCC] = 0.16-0.18, R2 = .32). Seven of the extant Ym models based on a linear regression approach also showed inaccurately estimated Ym values (root MSPE = 16.2%-36.0%, CCC = 0.02-0.27, R2 < .37). However, one of the developed models, which related to the complexity of the energy use efficiencies of the diet consumed to Ym, showed adequate accuracy at the farm level (root MSPE = 9.1%, CCC = 0.75, R2 = .67). Our results thus suggest a new Ym model and future challenges for estimating Zebu beef cattle production in tropical developing countries.


Microbial bioavailability of covalently bound polymer coatings on model engineered nanomaterials.
By controlling nanoparticle flocculation and deposition, polymer coatings strongly affect nanoparticle fate, transport, and subsequent biological impact in the environment. Biodegradation is a potential route to coating breakdown, but it is unknown whether surface-bound polymers are bioavailable. Here we demonstrate, for the first time, that polymer coatings covalently bound to nanomaterials are bioavailable. Model poly(ethylene oxide) (PEO) brush-coated nanoparticles (densely cross-linked bottle brush copolymers) with hydrophobic divinyl benzene cross-linked cores and hydrophilic PEO brush shells, having ~ 30 nm hydrodynamic radii, were synthesized to obtain a nanomaterial in which biodegradation was the only available coating breakdown mechanism. PEO-degrading enrichment cultures were supplied with either PEO homopolymer or PEO brush nanoparticles as the sole carbon source, and protein and CO₂ production were monitored as a measure of biological conversion. Protein production after 90 h corresponded to 14% and 8% of the total carbon available in the PEO homopolymer and PEO brush nanoparticle cultures, respectively, and CO₂ production corresponded to 37% and 3.8% of the carbon added to the respective system. These results indicate that the PEO in the brush is bioavailable. Brush biodegradation resulted in particle aggregation, pointing to the need to understand biologically mediated transformations of nanoparticle coatings in order to understand the fate and transport of nanoparticles in the environment.


Comparisons of three probability formulae for parentage exclusion.
Three general formulae calibrate the average capability of marker systems to dispute falsely reported pedigree records in uniparous species. The most familiar exclusion formula applies to paternity, although the same formula applies equally to maternity. Another formula faults the relationship of a single offspring with its putative parent; for example, where the genotype of the other parent is not available. The remaining formulae excludes both of the falsely recorded parents of a substituted offspring. Simplified forms of the three general formulae facilitate the calculation of maximal average exclusion values over a range of hypothetical markers. Allele frequency data on eight marker systems in horses provide practical examples. The exclusion values of the three formulae are compared.


Chronic inflammation and oxidative stress: the smoking gun for Helicobacter pylori-induced gastric cancer?
Helicobacter pylori is the leading risk factor associated with gastric carcinogenesis. H. pylori leads to chronic inflammation because of the failure of the host to eradicate the infection. Chronic inflammation leads to oxidative stress, deriving from immune cells and from within gastric epithelial cells. This is a main contributor to DNA damage, apoptosis and neoplastic transformation. Both pathogen and host factors directly contribute to oxidative stress, including H. pylori virulence factors, and pathways involving DNA damage and repair, polyamine synthesis and metabolism, and oxidative stress response. Our laboratory has recently uncovered a mechanism by which polyamine oxidation by spermine oxidase causes H 2O 2 release, DNA damage and apoptosis. Our studies indicate novel targets for therapeutic intervention and risk assessment in H. pylori-induced gastric cancer. More studies addressing the many potential contributors to oxidative stress, chronic inflammation, and gastric carcinogenesis are essential for development of therapeutics and identification of gastric cancer biomarkers.


Influence of Arabian Pharmacy on Diseases Tretament During Ottoman's Period in Bosnia and Herzegovina.
The Arab cultural heritage was an era of invaluable preservation and development of numerous teachings, including biomedical sciences. The golden period of Arab medicine deserves special attention in the history of medicine and pharmacy, as it was the period of rapid translation of works from Greek and Persian cultures into Arabic. They preserved their culture, and science from decay, and then adopted them to continue building their science on theirs as a basis. After the fall of Arabian Caliphate, Arabian pharmacy, continued to persevere, and spread through Turkish Caliphate until its fall in the First World War. That way, Arabian pharmacy will be spread to new areas that had benefited from it, including the area of occupied Bosnia and Herzegovina. Because of the vast territorial scope of the Ottoman Empire, the focus of this paper is description of developing pharmacy in Bosnia and Herzegovina during the time of Ottoman reign.


A role for alpha-synuclein in the regulation of dopamine biosynthesis.
The alpha-synuclein gene is implicated in the pathogenesis of Parkinson's disease. Although alpha-synuclein function is uncertain, the protein has homology to the chaperone molecule 14-3-3. In addition, alpha-synuclein can bind to 14-3-3, and both alpha-synuclein and 14-3-3 bind to many of the same proteins. Because 14-3-3 binds to and activates tyrosine hydroxylase, the rate-limiting enzyme in dopamine (DA) biosynthesis, we explored whether alpha-synuclein also bound to tyrosine hydroxylase and influenced its activity. Immunoprecipitation revealed an interaction between alpha-synuclein and tyrosine hydroxylase in brain homogenates and MN9D dopaminergic cells. Colocalization of alpha-synuclein with tyrosine hydroxylase was confirmed by immunoelectron microscopy. To explore the consequences of the interaction, we measured the effect of recombinant alpha-synuclein on tyrosine hydroxylase activity in a cell-free system and observed a dose-dependent inhibition of tyrosine hydroxylase by alpha-synuclein. To measure the impact of alpha-synuclein on tyrosine hydroxylase in dopaminergic cells, we stably transfected MN9D cells with wild-type or A53T mutant alpha-synuclein. Overexpression of wild-type or A53T mutant alpha-synuclein did not significantly alter tyrosine hydroxylase protein levels in our stably transfected cells. However, overexpressing cell lines had significantly reduced tyrosine hydroxylase activity and a corresponding reduction in dopamine synthesis. The reduction in cellular dopamine levels was not caused by increased dopamine catabolism or dopamine efflux. These data suggest that alpha-synuclein plays a role in the regulation of dopamine biosynthesis, acting to reduce the activity of tyrosine hydroxylase. If so, a loss of soluble alpha-synuclein, by reduced expression or aggregation, could increase dopamine synthesis with an accompanying increase in reactive dopamine metabolites.


Is specialization an evolutionary dead end? Testing for differences in speciation, extinction and trait transition rates across diverse phylogenies of specialists and generalists.
Specialization has often been claimed to be an evolutionary dead end, with specialist lineages having a reduced capacity to persist or diversify. In a phylogenetic comparative framework, an evolutionary dead end may be detectable from the phylogenetic distribution of specialists, if specialists rarely give rise to large, diverse clades. Previous phylogenetic studies of the influence of specialization on macroevolutionary processes have demonstrated a range of patterns, including examples where specialists have both higher and lower diversification rates than generalists, as well as examples where the rates of evolutionary transitions from generalists to specialists are higher, lower or equal to transitions from specialists to generalists. Here, we wish to ask whether these varied answers are due to the differences in macroevolutionary processes in different clades, or partly due to differences in methodology. We analysed ten phylogenies containing multiple independent origins of specialization and quantified the phylogenetic distribution of specialists by applying a common set of metrics to all datasets. We compared the tip branch lengths of specialists to generalists, the size of specialist clades arising from each evolutionary origin of a specialized trait and whether specialists tend to be clustered or scattered on phylogenies. For each of these measures, we compared the observed values to expectations under null models of trait evolution and expected outcomes under alternative macroevolutionary scenarios. We found that specialization is sometimes an evolutionary dead end: in two of the ten case studies (pollinator-specific plants and host-specific flies), specialization is associated with a reduced rate of diversification or trait persistence. However, in the majority of studies, we could not distinguish the observed phylogenetic distribution of specialists from null models in which specialization has no effect on diversification or trait persistence.


The cAMP phosphodiesterase Prune localizes to the mitochondrial matrix and promotes mtDNA replication by stabilizing TFAM.
Compartmentalized cAMP signaling regulates mitochondrial dynamics, morphology, and oxidative phosphorylation. However, regulators of the mitochondrial cAMP pathway, and its broad impact on organelle function, remain to be explored. Here, we report that Drosophila Prune is a cyclic nucleotide phosphodiesterase that localizes to the mitochondrial matrix. Knocking down prune in cultured cells reduces mitochondrial transcription factor A (TFAM) and mitochondrial DNA (mtDNA) levels. Our data suggest that Prune stabilizes TFAM and promotes mitochondrial DNA (mtDNA) replication through downregulation of mitochondrial cAMP signaling. In addition, our work demonstrates the prevalence of mitochondrial cAMP signaling in metazoan and its new role in mitochondrial biogenesis.


Localisation in focal epilepsy: a practical guide.
The semiology of epileptic seizures reflects activation, or dysfunction, of areas of brain (often termed the symptomatogenic zone) as a seizure begins and evolves. Specific semiologies in focal epilepsies provide an insight into the location of the seizure onset zone, which is particularly important for presurgical epilepsy assessment. The correct diagnosis of paroxysmal events also depends on the clinician being familiar with the spectrum of semiologies. Here, we summarise the current literature on localisation in focal epilepsies using illustrative cases and discussing possible pitfalls in localisation.


Geographical genetics of Pseudoplatystoma punctifer (Castelnau, 1855) (Siluriformes, Pimelodidae) in the Amazon Basin.
Geographical genetics allows the evaluation of evolutionary processes underlying genetic variation within and among local populations and forms the basis for establishing more effective strategies for biodiversity conservation at the population level. In this study, we used explicit spatial analyses to investigate molecular genetic variation (estimated using 7 microsatellite markers) of Pseudoplatystoma punctifer, by using samples obtained from 15 localities along the Madeira River and Solimões, Amazon Basin. A high genetic diversity was observed associated with a relatively low FST (0.057; P < 0.001), but pairwise FST values ranged from zero up to 0.21 when some pairs of populations were compared. These FST values have a relatively low correlation with geographic distances (r = 0.343; P = 0.074 by Mantel test), but a Mantel correlogram revealed that close populations (up to 80 km) tended to be more similar than expected by chance (r = 0.360; P = 0.015). The correlogram also showed a exponential-like decrease of genetic similarity with distance, with a patch-size of around 200 km, compatible with isolation-by-distance and analogous processes related to local constraints of dispersal and spatially structured levels of gene flow. The pattern revealed herein has important implications for establishing strategies to maintain genetic diversity in the species, especially considering the threats due to human impacts caused by building large dams in this river system.


Measuring carotid stenosis. Time for a reappraisal.
Data from recent multicenter carotid endarterectomy trials have questioned the validity and reliability of Doppler ultrasound in the assessment of carotid stenosis. We prospectively analyzed 45 patients undergoing carotid angiography to compare the North American Symptomatic Carotid Endarterectomy Trial (NASCET) and European Carotid Surgery Trial (ECST) methods of measuring carotid stenosis with those of direct visualization ("eyeballing") and duplex ultrasound. Linear NASCET and ECST measurements were also converted into area using the pi r2 function and termed "squared NASCET" (N2) and "squared ECST" (E2). In 15 of 45 patients undergoing carotid endarterectomy, the carotid plaque was removed intact, sectioned, and photographed for computer measurement of cross-sectional area. Comparison of this "gold standard" was then made to each method of measurement. Comparison between duplex and the various angiographic measurement techniques revealed significant differences between NASCET and duplex (P < .0001), ECST and duplex (P < .01), and E2 and duplex (P < .01) but not between N2, eyeballing, and carotid duplex methods. Even the NASCET and ECST methods themselves differed significantly (P < .006). When comparison was made with computerized planimetric measurements of the carotid plaque, there were significant differences for both NASCET (P < .0007) and ECST (P < .007). Correlation was demonstrated only between planimetry and N2, E2, and duplex. NASCET and ECST angiographic methods of measurement consistently underestimate the "true" anatomic stenosis. As such, they represent only "indexes" of carotid stenosis severity. Duplex provides a more accurate measurement of carotid stenosis.


Heterozygote deficiency, population substructure and their implications in DNA fingerprinting.
Substructured populations exhibit an overall deficiency of heterozygosity whose proportional magnitude depends on the nature of substructuring, i.e., the number of subpopulations (s), their time of divergence (t) from the ancestral population, and the rate of gene flow amongst them (m). Since apparent heterozygote deficiency could be caused by many factors other than population substructuring, one must examine the nature of substructuring that could produce the observed extent of heterozygote deficiency, in order to infer the substructuring from an observed heterozygote deficiency. Using the equivalence of proportional heterozygote deficiency and the coefficient of gene differentiation (GST), we can generate isolines of GST as functions of s, t (in units of 2Ne generations, Ne being the effective population size) and m. Analytical results suggest that large GST values cannot be reached by substructuring alone, unless the number of subpopulations are large and they remain isolated over a long period of time. Application of the theory to population data on six variable number of tandem repeats (VNTR) loci in US Caucasians and US Blacks demonstrates that the observed heterozygote deficiencies at these loci cannot be explained by substructuring within these populations alone. This is so because such large values of GST (3%-10%) would require an absence of gene exchange between the subpopulations and a divergence time from each other of at least 25,000 years ago, neither of which is compatible with the demography and ethnohistory of US Caucasians and Blacks. In contrast, the inability to detect extreme-sized alleles and/or incomplete resolution of nearly similar-sized alleles following Southern gel electrophoresis could easily explain the observed heterozygote deficiencies. The implications of these results are discussed in the context of the forensic use of DNA-typing data, and justify the employment of population genetic principles in forensic genetics.


Sustainability of a community-based anti-retroviral care delivery model - a qualitative research study in Tete, Mozambique.
To overcome patients' reported barriers to accessing anti-retroviral therapy (ART), a community-based delivery model was piloted in Tete, Mozambique. Community ART Groups (CAGs) of maximum six patients stable on ART offered cost- and time-saving benefits and mutual psychosocial support, which resulted in better adherence and retention outcomes. To date, Médecins Sans Frontières has coordinated and supported these community-driven activities. To better understand the sustainability of the CAG model, we developed a conceptual framework on sustainability of community-based programmes. This was used to explore the data retrieved from 16 focus group discussions and 24 in-depth interviews with different stakeholder groups involved in the CAG model and to identify factors influencing the sustainability of the CAG model. We report the findings according to the framework's five components. (1) The CAG model was designed to overcome patients' barriers to ART and was built on a concept of self-management and patient empowerment to reach effective results. (2) Despite the progressive Ministry of Health (MoH) involvement, the daily management of the model is still strongly dependent on external resources, especially the need for a regulatory cadre to form and monitor the groups. These additional resources are in contrast to the limited MoH resources available. (3) The model is strongly embedded in the community, with patients taking a more active role in their own healthcare and that of their peers. They are considered as partners in healthcare, which implies a new healthcare approach. (4) There is a growing enabling environment with political will and general acceptance to support the CAG model. (5) However, contextual factors, such as poverty, illiteracy and the weak health system, influence the community-based model and need to be addressed. The community embeddedness of the model, together with patient empowerment, high acceptability and progressive MoH involvement strongly favour the future sustainability of the CAG model. The high dependency on external resources for the model's daily management, however, can potentially jeopardize its sustainability. Further reflections are required on possible solutions to solve these challenges, especially in terms of human resources.


Purulent pericarditis after liver abscess: a case report.
We present the case of a 49-year-old woman, with previous clinical antecedents of recent hepatic metastasis, who was admitted to the ICU due to respiratory failure and hemodynamic instability. She was found to have purulent pericarditis complicated by pericardial tamponade and pleural effusion, as well as surgical site infection, which was the origin of the disease. Cultures of the surgical wound and the pericardial effusion were positive for Enterococcus faecalis and Escherichia coli. A pericardial tap was performed and the intra-abdominal abscess was surgically drained. Pleural effusion was also evacuated. She received antibiotic treatment and recovered successfully. The only after-effect was a well-tolerated effusive-constrictive pericarditis.


EDITORIAL.
This Supplement of the Journal Neuroendocrinology Letters presents papers from the 19th Interdisciplinary Toxicological Conference TOXCON 2014: Connecting for Safer Europe held in Stará Lesná, Slovakia from Septembert 24-26, 2014. This event was organized by the Slovak Toxicology Society SETOX, Bratislava, Slovakia, with the Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Bratislava, Slovakia. The scientific program of the conference involved various subjects of experimental, clinical and industrial toxicology. The program was divided into individual sections dealing with specific topics of in vivo and in vitro studies, such as cellular and organ toxicity, genotoxicity, cytotoxicity, mutagenicity, developmental toxicology, military toxicology, ecotoxicology, methodological approaches in toxicology as well as problems of relative safety assessment of drugs and xenobiotics, alternative methods in toxicology, ethical problems and legislation. The meeting attendance exceeded 170 scientists from 8 countries. The presence of outstanding scientists giving keynote lectures provided an opportunity to familiarize with up-to-date research in various fields of toxicology and related subjects. Out of the 143 presentations given at the Conference, the outstanding 29 selected by the scientific committee, are highlighted and documented in this Supplement.


The gamma-tubulin gene family in humans.
Despite the central role of gamma-tubulin in the organization of the microtubule cytoskeleton, the gamma-tubulin gene family in humans has not been characterized. We now report the identification of a second expressed human gamma-tubulin gene (TUBG2) and a gamma-tubulin pseudogene (TUBG1P) in addition to the previously identified gamma-tubulin gene (TUBG1). Evidence from Southern hybridizations suggests that there are probably no additional gamma-tubulin sequences in the human genome. TUBG1 and TUBG2 are within 20 kb of each other in region q21 of chromosome 17, and TUBG1P is on chromosome 7. The proteins encoded by TUBG1 and TUBG2 share 97.3% amino acid identity, and the two genes are coexpressed in a variety of tissues. Previous studies of gamma-tubulin in human tissues and cell lines have been based on the tacit assumption that a single gamma-tubulin (the gamma-tubulin encoded by TUBG1) was present. While this assumption is not correct, the similarity of the products of TUBG1 and TUBG2 suggests that results of previous immunolocalization and immunoprecipitation studies in human cells and tissues are likely to be valid. In addition, any pharmacological agents that target one human gamma-tubulin are likely to target both.


The role of vitamin B12 in fasting hyperhomocysteinemia and its interaction with the homozygous C677T mutation of the methylenetetrahydrofolate reductase (MTHFR) gene. A case-control study of patients with early-onset thrombotic events.
Total fasting plasma homocysteine (tHcy), homozygosity for the C677T mutation of the methylenetetrahydrofolate reductase (MTHFR) gene and for the A2756G mutation of the methionine synthase (MS) gene, vitamin B12 and folate plasma levels were evaluated in 170 consecutive patients (89 M, 81 F; mean age 41 +/- 12 yrs) with documented early-onset thrombosis (89 venous, 69 arterial, 12 both; mean age at first episode 36 +/- 11 yrs), and in 182 age- and sex-matched healthy control subjects. Moderate hyperhomocysteinemia (HHcy, tHcy >19.5 microM in men and >15 microM in women) was detected in 45 patients (26.5%) and in 18 controls (9.9%, Mantel-Haenszel OR and 95% C.I. after stratification for arterial or venous thrombosis: 3.25, 1.78-5.91). The 677TT MTHFR genotype was not significantly more prevalent in patients (27.6%) than in controls (21.4%, RR = 1.42: 0.84-2.41), and markedly contributed to HHcy (Mantel-Haenszel RR after stratification for case/control status: 8.29, 4.61-14.9). The 2756GG MS genotype, observed in 4 patients (2.4%) and 8 controls (4.4%), was not associated to HHcy. tHcy was negatively correlated to folate and vitamin B12 levels, with better correlation found in subjects with the 677TT mutation (r = -0.42 and -0.25) than with the 677CC or CT MTHFR genotype (r = 0).37 and -0.11). However, folate was similar in patients and controls and vitamin B12 was higher in patients (460 +/- 206 vs. 408 +/-185 pg/ml, p = 0.011). In a generalized linear model, 44% of the variation in tHcy levels was explained by folate and vitamin B12 levels, the MTHFR genotype, gender, and by the interaction of the MTHFR genotype with folate (p < or =0.028); the interactions of vitamin B12 with the MTHFR genotype, gender and patient/control status also significantly contributed to the variation in tHcy levels (p < or =0.028). A 4-week administration of 5-methyltetrahydrofolate (15 mg/day) markedly lowered plasma tHcy in 24 patients with MTHFR 677TT genotype, but the response to treatment correlated with vitamin B,2 levels (p = 0.023). Subjects carrying the MTHFR 677TT genotype have higher folate and vitamin B12 requirements irrespective of the A2756G polymorphism of the MS gene. Yet unidentified abnormalities of MS or of any of the enzymes participating in the synthesis of methylated vitamin B12 may play an important role in the phenotypic expression of moderate hyperhomocysteinemia.


Influence of Tobacco Smoking on Perioperative Risk of Venous Thromboembolism.
Venous thromboembolism (VTE) is generally considered a process very different from arterial atherosclerosis. The role of tobacco is well known in arterial thrombosis. However, its role in VTE is less obvious and remains controversial. In this mini review, we analysed the literature to identify the role of active or passive smoking in perioperative VTE and the relationship between arterial atherosclerosis and VTE. We carried an Internet search in French and English including the following keywords: deep vein thrombosis, tobacco, cigarette smoking, pulmonary embolism, postoperative, postoperative, atherosclerosis. Regarding the relationship between tobacco and VTE, a total of 9 studies were identified. Studies were ranked by the risk of VTE in relation to active or passive smoking. In conclusion, the management of smoking during the perioperative period for a short-term arrest (minimum 4-8 weeks before the intervention), or long term, allows among others a reduction of arterial or venous thrombotic events. However, it is clear that the training of anaesthesiologists in the management of smoking will contribute to the reduction of this public health problem.


Renal colic, where is it headed? An observational study.
In the last thirty years, the treatment for renal and ureteral calculi has undergone profound variations. The objective of this study has been to evaluate the existence of parameters which can affect the spontaneous expulsion of a symptomatic ureteral stone in a reasonably brief period of time and to identify whether certain parameters such as sex, age, the location and dimension of the stone, the presence of dilation in the urinary tract together with the administered therapy, can be used for a correct clinical management of the patient. In a period of 9 months, 486 cases of renal colic were registered at emergency department. The cases of renal colic due to ureteral calculus were 188 (38.7%). The patients' charts, complete of all data and therefore, valid for this research, resulted to be 120 (64%). In the presence of a symptomatic ureteral stone, the correct approach must first of all, focalize on the dimension of the calculus itself; less importance instead, is given to the location, as reported in other studies, the presence of hydroureteronephrosis, sex and the side. In the cases when the pain symptoms cannot be solved by means of the administration of analgesics, it is then reasonable to take into consideration an immediate endourological treatment. If the pain symptoms are promptly solved, an attentive wait of 4 weeks should be considered reasonable in order to allow spontaneous expulsion of the calculus.


The evaluation of effect of developer age in the detection of approximal caries using three speed dental x-ray films: an in-vitro study.
Ever since the first radiograph was taken in 1895, radiographs have become an essential part of dental practice. Since radiation exposure for the patients is a major concern, it is important for us to reduce the radiation exposure to the patient and surrounding environment, without compromising the quality of image. Since improper film processing is one of the major causes of poor image quality, it is very important to test dental films under a variety of processing conditions. So this study was conducted to compare the diagnostic accuracy between Kodak Ultraspeed, Ektaspeed and Agfa Dentus M2 Comfort films for the detection of approximal caries in fresh as well as in aged processing solution. Hundred extracted unrestored maxillary and mandibular teeth were aligned in a group of five teeth in each plaster model. Total of 20 plaster models were constructed and only 3 teeth in the middle were utilized for the study. The final sample consisted of 60 teeth in the study. The exposure time for Ultraspeed film was standardized at 0.5 seconds and for Ektaspeed and Agfa Dentus M2 films, the exposure time was 0.32 seconds. For each radiograph two blocks were used simulating bitewing radiograph. All 20 plaster blocks were exposed using three film types at standardized exposure timings with 30 radiographs each week. The procedure was repeated every week until 6 weeks. Radiographs were evaluated by an observer and assessed by following four point scale 0- sound tooth, 1- lesion in enamel, 2- lesion in amelodentinal junction, 3- dentinal lesion. Only distal surfaces were assessed. Thereafter all the teeth were subjected for histological study and actual depth of the lesion was recorded which acted as gold standard and this reading was subtracted in observer readings after radiographic examination. All data collected were analyzed statistically using ANOVA, paired and unpaired t-tests. Ektaspeed and Agfa films required less exposure compared to Ultraspeed film. With respect to film, all films had equal diagnostic accuracy upto three weeks. From fourth week Ultraspeed and Agfa films were considerably better than E speed film. E speed film was inferior in the depleted solution compared to other two solutions. With respect to lesion detection Agfa film was inferior to other two films. All three films had equal diagnostic accuracy in fresh solution. Considering the significant reduction in patient radiation with Ektaspeed film, this should be the film of choice whenever possible. As far as the diagnosis of approximal caries is concerned processing solutions should not be used longer than 3 weeks.


Neuroendocrine regulation of stimulated ACTH secretion.
Two models of corticosteroid inhibition of stimulus-induced ACTH secretion are discussed. Modulation of ACTH secretion by a nonsteroidal factor related to adrenal mass is reviewed. Experiments are presented that compare feedback inhibition of ACTH affected by corticosterone and by adrenal mass. Use of two stimuli to ACTH secretion and manipulation of two feedback signals allow the conclusion that only one of the steroid feedback models will satisfy the data, and that corticosteroid feedback inhibition of stimulus-induced ACTH secretion must be exerted at at least two sites in the central nervous system.


Genomewide evidence of environmentally mediated secondary contact of European green crab (Carcinus maenas) lineages in eastern North America.
Genetic-environment associations are increasingly revealed through population genomic data and can occur through a number of processes, including secondary contact, divergent natural selection, or isolation by distance. Here, we investigate the influence of the environment, including seasonal temperature and salinity, on the population structure of the invasive European green crab (Carcinus maenas) in eastern North America. Green crab populations in eastern North America are associated with two independent invasions, previously shown to consist of distinct northern and southern ecotypes, with a contact zone in southern Nova Scotia, Canada. Using a RAD-seq panel of 9,137 genomewide SNPs, we detected 41 SNPs (0.49%) whose allele frequencies were highly correlated with environmental data. A principal components analysis of 25 environmental variables differentiated populations into northern, southern, and admixed sites in concordance with the observed genomic spatial structure. Furthermore, a spatial principal components analysis conducted on genomic and geographic data revealed a high degree of global structure (p < .0001) partitioning a northern and southern ecotype. Redundancy and partial redundancy analyses revealed that among the environmental variables tested, winter sea surface temperature had the strongest association with spatial structuring, suggesting that it is an important factor defining range and expansion limits of each ecotype. Understanding environmental thresholds associated with intraspecific diversity will facilitate the ability to manage current and predict future distributions of this aquatic invasive species.


Inhibition of cyclic strain-induced endothelin-1 secretion by tetramethylpyrazine.
1. Chuanxiong is a Chinese herb that has been used widely in China to treat vascular disorders. 2,3,5,6-Tetramethylpyrazine (TMP) is one of the major components purified from chuanxiong. Many studies have demonstrated that TMP is effective in the treatment of cardiovascular diseases. However, the mechanism of action by which TMP exerts relaxation in vascular vessels remains unclear. 2. Endothelin (ET)-1 is a potent vasopressor synthesised by endothelial cells both in culture and in vivo. The aims of the present study were to test the hypothesis that TMP may alter strain-induced ET-1 secretion and to identify the putative underlying signalling pathways in endothelial cells. 3. We showed that TMP inhibits strain-induced ET-1 secretion. 2,3,5,6-Tetramethylpyrazine also inhibits the strain-induced formation of reactive oxygen species (ROS) and phosphorylation of extracellular signal-regulated kinases (ERK) 1/2. Furthermore, pretreating cells with TMP or the anti-oxidant N-acetyl-cysteine decreased strain-induced increases in ET-1 secretion and ERK1/2 phosphorylation. Using a reporter gene assay, TMP and N-acetyl-cysteine were demonstrated to also attenuate the strain-induced activity of the activator protein-1 reporter. 4. In summary, we have demonstrated, for the first time, that TMP inhibits strain-induced ET-1 gene expression, in part by interfering with the ERK1/2 pathway via attenuation of ROS formation. Thus, the present study provides important new insights into the molecular pathways that may contribute to the proposed beneficial effects of TMP in the vascular system.