Oleosins (24 and 18 kDa) are hydrolyzed not only in extracted soybean oil bodies but also in soybean germination.
After oil bodies (OBs) were extracted from ungerminated soybean by pH 6.8 extraction, it was found that 24 and 18 kDa oleosins were hydrolyzed in the extracted OBs, which contained many OB extrinsic proteins (i.e., lipoxygenase, β-conglycinin, γ-conglycinin, β-amylase, glycinin, Gly m Bd 30K (Bd 30K), and P34 probable thiol protease (P34)) as well as OB intrinsic proteins. In this study, some properties (specificity, optimal pH and temperature) of the proteases of 24 and 18 kDa oleosins and the oleosin hydrolysis in soybean germination were examined, and the high relationship between Bd 30K/P34 and the proteases was also discussed. The results showed (1) the proteases were OB extrinsic proteins, which had high specificity to hydrolyze 24 and 18 kDa oleosins, and cleaved the specific peptide bonds to form limited hydrolyzed products; (2) 24 and 18 kDa oleosins were not hydrolyzed in the absence of Bd 30K and P34 (or some Tricine-SDS-PAGE undetectable proteins); (3) the protease of 24 kDa oleosin had strong resistance to alkaline pH while that of 18 kDa oleosin had weak resistance to alkaline pH, and Bd 30K and P34, resolved into two spots on two-dimensional electrophoresis gel, also showed the same trend; (4) 16 kDa oleosin as well as 24 and 18 kDa oleosins were hydrolyzed in soybean germination, and Bd 30K and P34 were always contained in the extracted OBs from germinated soybean even when all oleosins were hydrolyzed; (5) the optimal temperature and pH of the proteases were respectively determined as in the ranges of 35-50 °C and pH 6.0-6.5, while 60 °C or pH 11.0 could denature them.


Autolytic activity and molecular characteristics of Staphylococcus haemolyticus strains with induced vancomycin resistance.
The aim of this study was to investigate the molecular characteristics of induced vancomycin resistance in Staphylococcus haemolyticus. Autolytic properties and phenotypic characteristics of passage-selected vancomycin-resistant S. haemolyticus strains were examined. In addition, expression of autolysis-related genes (atl, lrgAB, sarA and lytS) was investigated using the RNase protection assay (RPA). The RPA results indicated that only the expression of the atl gene was significantly upregulated (2.5- to 6-fold increase) in vancomycin-intermediate and vancomycin-resistant strains. The vancomycin-resistant strains exhibited lower expression of murein hydrolase proteins and reduced autolytic activity compared with the parent strain. In addition, a reduced growth rate, cell wall thickening and higher survival rate in the presence of lysostaphin were observed in vancomycin-intermediate and vancomycin-resistant induced strains compared with the parent strain. In conclusion, altered autolytic properties, in particular upregulation of the atl gene, may contribute to vancomycin resistance in S. haemolyticus.


Hydrazones in anion transporters: the detrimental effect of a second binding site.
Synthetic anion transporters can be developed using anion receptors that are able to bind the anion and stabilize it in the lipophilic interior of a bilayer membrane, and they usually contain functional groups with acidic NHs, such as ureas, thioureas and squaramides. To assess the suitability of acylhydrazones as a new functional group for the preparation of anion transporters, we have studied a family of thioureas functionalized with these and related functional groups. 1H NMR titrations and DFT calculations indicate that the thioureas bearing acylhydrazone groups behave as chloride receptors with two separate binding sites, of which the acylhydrazone binds weaker than the thiourea. Chloride transport studies show that the additional binding site has a detrimental effect on thiourea-based transporters, and this phenomenon is also observed for bis(thio)ureas with two separate binding sites. We propose that the presence of a second anion binding unit hinders the transport activity of the thiourea due to additional interactions with the phospholipids of the membrane. In agreement with this hypothesis, extensive molecular dynamics simulations suggest that the molecules will tend to be positioned in the water/lipid interface, driven by the interaction of the NHs of the thiourea and of the acylhydrazone groups with the POPC polar head groups and water molecules. Moreover, the interaction energies show that the poorest transporters have indeed the strongest interactions with the membrane phospholipids, inhibiting chloride transport. This detrimental effect of additional functional groups on transport activity should be considered when designing new ion transporters, unless these groups cooperatively promote anion recognition and transmembrane transport.


Post-Transplant Diabetes Mellitus After Kidney Transplant in Hispanics and Caucasians Treated with Tacrolimus-Based Immunosuppression.
BACKGROUND Development of post-transplant diabetes mellitus after kidney transplant (PTDM) significantly increases kidney graft loss and mortality. Several risk factors for PTDM have been reported, including Hispanic ethnicity and the use of calcineurin inhibitors and corticosteroids. The incidence and impact of PTDM in the Hispanic kidney transplant population is unknown. MATERIAL AND METHODS We retrospectively reviewed the medical records of 155 Hispanic and 124 Caucasian patients, who were not diabetics and underwent kidney transplant between January 2006 and December 2011. We analyzed their clinical outcomes at 12 months post-transplant, including the incidence of PTDM, acute rejection rates, and patient and graft survival. RESULTS Hispanics who developed PTDM (n=22) were more than 10 years older and had higher body mass index (BMI) than Hispanics without PTDM (p<0.001 and p=0.001, respectively). Caucasians with PTDM (n=13) were non-significantly older (2.5 years) and had higher BMI than Caucasians without PTDM (p=0.526, p=0.043, respectively). The incidence of PTDM was not significantly different between Hispanics and Caucasians treated with tacrolimus-based immunosuppression (14.2% and 10.5%, respectively). CONCLUSIONS PTDM did not cause significant difference in short-term outcomes after kidney transplant in Hispanics or Caucasians. Larger multicenter prospective and long-term clinical trials are needed to validate these findings.


IL-6-independent association of elevated serum neopterin levels with prevalent frailty in community-dwelling older adults.
neopterin is a monocyte/macrophage-derived immune activation marker and its levels increase with age. Frailty is an important clinical syndrome of old age. Previous studies have shown significant association between elevated interleukin-6 (IL-6) levels and frailty. The objective of this study was to evaluate IL-6-independent association of serum neopterin levels with prevalent frailty. this is a cross-sectional study in community-dwelling older adults recruited from residential and retirement communities in Baltimore, MD, USA. Frailty was determined using validated screening criteria. Serum neopterin and IL-6 levels were measured using standard enzyme-linked immunosorbent assay. Pearson correlation and multivariate linear regression analysis was performed to assess the relationship between log(neopterin) and log(IL-6). Odds ratios (ORs) for frailty were calculated using log(neopterin) and log(IL-6) as continuous measures and across tertiles of neopterin and IL-6 levels, adjusting for age, race, sex, education and body mass index. one hundred and thirty-three individuals with a mean age of 84 years (range 72-97) completed the study. Neopterin levels were significantly higher in frail older adults than those in non-frail controls [median: 8.94 versus 8.35 nM, respectively, P < 0.001 t-test on log(neopterin)]. Log(neopterin) was significantly associated with prevalent frailty, adjusting for log(IL-6). Participants in the top tertile of neopterin had OR of 3.80 [95% confidence interval (CI) = 1.36-10.6, P < 0.01] for frailty. As expected, participants in the top tertile of IL-6 had OR of 3.29 (95% CI = 1.21-7.86, P < 0.05) for frailty. Log(neopterin) correlated with log(IL-6) (correlation coefficient = 0.19, P < 0.05). Moreover, OR for participants in the top neopterin tertile remained significant after adjusting for IL-6 (OR = 3.97, 95% CI = 1.15-13.72, P < 0.05). elevated neopterin levels had IL-6-independent association with prevalent frailty, suggesting potential monocyte/macrophage-mediated immune activation in the frail elderly.


[The truth about the clinical reality is in the registries. A register-based randomized clinical trial could provide representative picture].
A randomized, controlled clinical trial (RCT) is the best way to compare two treatment options since it eliminates the problem with confounding. However, todays' RCTs are often limited by including selected patients by the use of narrow inclusion criteria and multiple exclusion criteria. In a recent study of myocardial infarction survivors, the median age was 10 years older and the long term risk was 2-3 times higher in »real-world« data from a national registry compared with results from recently performed RCTs. These results raise questions about the generalizability of many RCTs. Our registries should therefore be used more often to evaluate new treatments. Registry-based randomized, controlled clinical trial (R-RCT) is a new concept that may not only lower the costs of randomized studies but also increase the generalizability of the trials.


Cofactor-dependent conformational heterogeneity of GAD65 and its role in autoimmunity and neurotransmitter homeostasis.
The human neuroendocrine enzyme glutamate decarboxylase (GAD) catalyses the synthesis of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) using pyridoxal 5'-phosphate as a cofactor. GAD exists as two isoforms named according to their respective molecular weights: GAD65 and GAD67. Although cytosolic GAD67 is typically saturated with the cofactor (holoGAD67) and constitutively active to produce basal levels of GABA, the membrane-associated GAD65 exists mainly as the inactive apo form. GAD65, but not GAD67, is a prevalent autoantigen, with autoantibodies to GAD65 being detected at high frequency in patients with autoimmune (type 1) diabetes and certain other autoimmune disorders. The significance of GAD65 autoinactivation into the apo form for regulation of neurotransmitter levels and autoantibody reactivity is not understood. We have used computational and experimental approaches to decipher the nature of the holo → apo conversion in GAD65 and thus, its mechanism of autoinactivation. Molecular dynamics simulations of GAD65 reveal coupling between the C-terminal domain, catalytic loop, and pyridoxal 5'-phosphate-binding domain that drives structural rearrangement, dimer opening, and autoinactivation, consistent with limited proteolysis fragmentation patterns. Together with small-angle X-ray scattering and fluorescence spectroscopy data, our findings are consistent with apoGAD65 existing as an ensemble of conformations. Antibody-binding kinetics suggest a mechanism of mutually induced conformational changes, implicating the flexibility of apoGAD65 in its autoantigenicity. Although conformational diversity may provide a mechanism for cofactor-controlled regulation of neurotransmitter biosynthesis, it may also come at a cost of insufficient development of immune self-tolerance that favors the production of GAD65 autoantibodies.


Integrated Metabolomics, Transcriptome and Functional Analysis Reveal Key Genes Are Involved in Tree Age-Induced Amino Acid Accumulation in Torreya grandis Nuts.
Torreya grandis is native Chinese tree species of economic significance, renowned for its long lifespan and the rich nutritional value of its nuts. In this study, we analyzed the morphological characteristics, metabolites, associated gene expressions, and regulatory mechanism in nuts from young (10 years old) and old (1000 years old) T. grandis trees. We observed that the length, width, and weight of nuts from older trees were considerably greater than those from younger trees. Metabolomic analysis revealed that the concentrations of 18 amino acids and derivatives (including histidine and serine) in nuts from older trees were markedly higher than those in nuts from younger trees. Transcriptome and metabolomic correlation analysis identified 16 genes, including TgPK (pyruvate kinase), TgGAPDH (glyceraldehyde 3-phosphate dehydrogenase), and others, which exhibit higher expression levels in older trees compared to younger trees, as confirmed by qRT-PCR. These genes are associated with the biosynthesis of histidine, glutamic acid, tryptophan, and serine. Transient expression of TgPK in tobacco led to increased pyruvate kinase activity and amino acid content (histidine, tryptophan, and serine). Additionally, dual-luciferase assays and yeast one-hybrid results demonstrated that TgWRKY21 positively regulates TgPK expression by directly binding to the TgPK promoter. These findings not only demonstrate the nutritional differences between nuts from young and old trees but also offer fresh insights into the development of nutritional sources and functional components based on nuts from old trees, enriching our understanding of the potential benefits of utilizing nuts from older trees.


Ketogenic diet in the treatment of epilepsy in children under the age of 2 years: study protocol for a randomised controlled trial.
The incidence of epilepsy is greatest in the first 2 years of life, an age group where there is generally a poor prognosis for both seizure control and neurodevelopmental outcome. Early control of seizures can be associated with better developmental outcome but many of the epilepsies presenting in infancy are poorly responsive to antiepileptic medication. The ketogenic diet (KD) is a high-fat, low-carbohydrate diet designed to mimic the effects of starvation on the body. Dietary fat is converted into ketones in the body and used as an energy source by the brain. The KD has been shown to be successful in controlling seizures in many observational studies, and in two randomised controlled trials (RCTs) in older children. However, little evidence is available in the very young. An open-label RCT where eligible children (age 3 months to 2 years with epilepsy who have failed two antiepileptic drugs (AEDs)) undergo baseline assessment, including medical and seizure history. Participants then start an observation period (7 or 14 days) with documentation of seizure frequency. Randomisation will occur on day 8 or day 15 to receive the KD or a further AED; the allocated treatment will commence on day 15, with instruction and training. A second assessment (4 weeks after start of treatment) will include a clinical review and tolerability questionnaire (modified Hague Scale of Side Effects - for those allocated to the KD group). Assessments will be repeated at 8 weeks after the start of treatment including biochemical investigations, after which, according to patient response, KD (diet group) or AED (standard AED group) will then be continued or changed. Those in the AED group who have failed to achieve seizure control at the 8-week assessment will then be offered KD outside the context of the trial. Those in the KD arm who fail to achieve seizure control will be changed to standard clinical management. All patients will be followed up for 12 months from randomisation for retention, seizure outcome, quality of life and neurodevelopmental status. The slow rate of recruitment is an ongoing practical issue. There is a limitation to the number of eligible patients compared to what was predicted, mainly due to the nature of this patient group. After a substantial amendment to widen inclusion criteria and reduce the baseline period to 7 days for patients with a high seizure burden, the rate of recruitment steadily increased. A number of operational concerns regarding dietetic time were also highlighted impacting on the recruitment rate. However, the combination of a low dropout rate and the opening of further centres, the trial should successfully meet the final recruitment target. All nine centres are now recruiting and we hope to open further centres within the UK. ClinicalTrials.gov, identifier: NCT02205931 . Registered on 16 December 2013.


Can a coarse-grained water model capture the key physical features of the hydrophobic effect?
Coarse-grained (CG) molecular dynamics can be a powerful method for probing complex processes. However, most CG force fields use pairwise nonbonded interaction potentials sets, which can limit their ability to capture complex multi-body phenomena such as the hydrophobic effect. As the hydrophobic effect primarily manifests itself due to the nonpolar solute affecting the nearby hydrogen bonding network in water, capturing such effects using a simple one CG site or "bead" water model is a challenge. In this work, we systematically test the ability of CG one site water models for capturing critical features of the solvent environment around a hydrophobe as well as the potential of mean force (PMF) of neopentane association. We study two bottom-up models: a simple pairwise (SP) force-matched water model constructed using the multiscale coarse-graining method and the Bottom-Up Many-Body Projected Water (BUMPer) model, which has implicit three-body correlations. We also test the top-down monatomic (mW) and the Machine Learned mW (ML-mW) water models. The mW models perform well in capturing structural correlations but not the energetics of the PMF. BUMPer outperforms SP in capturing structural correlations and also gives an accurate PMF in contrast to the two mW models. Our study highlights the importance of including three-body interactions in CG water models, either explicitly or implicitly, while in general highlighting the applicability of bottom-up CG water models for studying hydrophobic effects in a quantitative fashion. This assertion comes with a caveat, however, regarding the accuracy of the enthalpy-entropy decomposition of the PMF of hydrophobe association.


Pursuing passionate professionalism: the journey of one ophthalmic nurse. Part 2.
The final question to be asked: Is nursing a profession worth pursuing with passion in the 21st century? Florence Nightingale left a legacy rich in values, ethics, and standards that still shines light on the paths of 21st century nurses. Florence Nightingale pursued her passion with abandon. She held many positions in her career, but she is remembered for her story. Thank you, Florence Nightingale, for laying the solid foundation of courage, character, and commitment for all nursing generations to come. To this question, I, as an ophthalmic nurse, must answer a resounding yes, and I say that this noble profession is still worth pursuing with passion and commitment. What we nurses do has eternal value. When we pursue our role with passion, character, and integrity, we will influence future generations. We may not be remembered by our titles, but each life we touch will remember our story.


Immunogenic Effect of Hyperthermia on Enhancing Radiotherapeutic Efficacy.
Hyperthermia is a cancer treatment where tumor tissue is heated to around 40 °C. Hyperthermia shows both cancer cell cytotoxicity and immune response stimulation via immune cell activation. Immunogenic responses encompass the innate and adaptive immune systems, involving the activation of macrophages, natural killer cells, dendritic cells, and T cells. Moreover, hyperthermia is commonly used in combination with different treatment modalities, such as radiotherapy and chemotherapy, for better clinical outcomes. In this review, we will focus on hyperthermia-induced immunogenic effects and molecular events to improve radiotherapy efficacy. The beneficial potential of integrating radiotherapy with hyperthermia is also discussed.


Absence of genetic linkage of chromosome 5q31 with asthma and atopy in the general population.
Clinical asthma is associated with increased serum total immunoglobulin E (IgE), atopy (skin prick test positivity to common aeroallergens), and bronchial hyperreactivity (BHR) to non-specific stimuli (positive methacholine challenge test). A region on chromosome 5q31-33 has been linked with increased total serum IgE and BHR. A study of the genetic linkage of this region with clinical asthma and atopy was therefore undertaken. A polymorphic microsatellite marker in chromosome 5q31 (D5S399) was studied in 119 sibling pairs recruited from the general population who shared asthma, atopy, and/or BHR. Based on our population distribution of 13 different alleles, it was expected that by chance alone sibling pairs would share on average 1.24 alleles and that a significant excess would indicate genetic linkage. No evidence of linkage was found in 45 siblings concordant for asthma (shared alleles = 1.09, p = 0.95), in 103 sibling pairs with atopy (shared alleles = 1.18, p = 0.82), in 51 sibling pairs with BHR (shared alleles = 1.22, p = 0.62), or in 68 sibling pairs who shared atopy in the absence of BHR (shared alleles = 1.22, p = 0.61). A slight non-significant excess of shared alleles (1.44, p = 0.11) was observed in siblings who shared BHR without atopy. No evidence of genetic linkage of chromosome 5q31 with either clinical asthma or atopy was therefore detected in the population studied. Linkage between chromosome 5q and BHR needs further investigation.


Fatty acid analogue accumulation: a marker of myocyte viability in ischemic-reperfused myocardium.
A 3-methyl substituted radioiodinated long chain fatty acid analogue was evaluated as an agent for the noninvasive detection of altered fatty acid uptake in reperfused, postischemic myocardium. This iodinated fatty acid analogue, 15-(para-iodophenyl)-3-methyl pentadecanoic acid, was given intravenously at 3 hours of reperfusion following 15 minutes (Group 1, n = 5 dogs) or 60 minutes (Group 2, n = 5 dogs) of left anterior descending coronary artery occlusion. Myocardial blood flow (MBF) was measured during occlusion and reperfusion with radiolabeled microspheres administered via the left atrium. Paired ultrasonic subendocardial crystals were placed in the ischemic perfusion bed to assess regional left ventricular systolic function at baseline, during ischemia and reperfusion. Electron microscopic analysis and staining with triphenyltetrazolium chloride (TTC) was performed. Groups 1 and 2 dogs had similar (p = NS) myocardial blood flows during occlusion. TTC positive 1 g endocardial segments from Group 1 (n = 98) and Group 2 (n = 71) had 37% greater fatty acid analogue activity (0.26 +/- 0.04 vs. 0.19 +/- 0.09 percent injected dose per gram; p less than 0.05) compared with TTC negative segments from Group 2 dogs (n = 37). When fatty acid analogue activity was related to near simultaneous reperfusion blood flow, this ratio was 27% greater (p less than 0.05) in TTC positive segments (0.38 +/- 0.1) compared with TTC negative (0.30 +/- 0.16) segments, and 9% greater than normal (0.35 +/- 0.09; p less than 0.05). While ischemic regions from both Groups 1 and 2 dogs became similarly dyskinetic during occlusion (systolic shortening, -11 +/- 6 vs. -11 +/- 2%; p = NS), TTC negative segments remained akinetic (= 1 +/- 7%) at 3 hours of reperfusion while TTC positive zones had recovered partial systolic function (8 +/- 22%). Electron microscopy confirmed the presence of reversible ultrastructural changes in TTC positive regions. A 60-minute occlusion, 3-hour reperfusion model adapted for in vivo single photon emission computed tomography showed a similar excess of 123I fatty acid activity over flow when compared to perfusion (as measured with 201Tl) in the ischemic border zone of 4/4 canine myocardial infarcts. We conclude that the accumulation of this non-beta-oxidized fatty acid analogue noninvasively identifies zones of discordance between fatty acid and flow distribution that are characteristic of ischemically "stunned" but viable myocardium.


Membrane solubilization in erythrocytes as a measure of radiation exposure to fast neutrons.
Membrane solubilization and osmotic fragility of rat erythrocytes irradiated in vivo with fast neutron fluences ranging from 10(6) to 5 x 10(7) n cm(-2) using a 252Cf source were measured instantaneously using a light scattering technique. The solubilization of erythrocyte membrane by a non-ionic detergent, octylglucoside (OG), was found to exhibit a two stage transition from vesicular form to mixed micellar form in the range of detergent concentrations 1.5-7.8 mM. The coexistence phase, vesicular/mixed micellar, was shifted towards higher detergent concentrations with increase in the neutron fluence, indicating increasing membrane resistance to the detergent and hence change in the natural membrane permeation properties. The technique shows an adequate sensitivity in detecting membrane damage in erythrocytes and has potential as a biophysical marker of radiation exposure. The osmotic fragility of irradiated erythrocytes shows a decreasing trend with increasing irradiation fluence measured directly and two weeks post-irradiation. Blood films photographed two weeks post-irradiation show developed elliptocytosis and crenated cell anaemia.


[Social determinants of odontalgia in epidemiological studies: theoretical review and proposed conceptual model].
The epidemiological literature has been limited by the absence of a theoretical framework reflecting the complexity of causal mechanisms for the occurrence of health phenomena / disease conditions. In the field of oral epidemiology, such lack of theory also prevails, since dental caries the leading topic in oral research has been often studied through a biological and reductionist viewpoint. One of the most important consequences of dental caries is dental pain (odontalgia), which has received little attention in studies with sophisticated theoretical models and powerful designs to establish causal relationships. The purpose of this study is to review the scientific literature on the determinants of odontalgia and to discuss theories proposed for the explanation of the phenomenon. Conceptual models and emerging theories on the social determinants of oral health are revised, in an attempt to build up links with the bio-psychosocial pain model, proposing a more elaborate causal model for odontalgia. The framework suggests causal pathways between social structure and oral health through material, psychosocial and behavioral pathways. Aspects of the social structure are highlighted in order to relate them to odontalgia, stressing their importance in discussions of causal relationships in oral health research.


Prevalence of dry eye during the COVID-19 pandemic: A systematic review and meta-analysis.
During the COVID-19 pandemic, many people devoted longer time to screen viewing due to the need for study, work, and online social activities, instead of outdoor activities, which may have led to an increase in dry eye symptoms. This study aimed to evaluate the prevalence of dry eye during the COVID-19 pandemic. PubMed, Cochrane Library, Embase, and Web of Science were searched from January 1, 2020 to October 20, 2022. Cross-sectional surveys on dry eye prevalence conducted after January 1, 2020 were included. Two review authors independently performed data extraction and assessed study quality. The random-effects model was used to analyze the prevalence of dry eye, and the odds ratio was used to assess the strength of the association between variables. Subgroup analysis was performed to detect heterogeneity, the leave-one-out method for sensitivity analysis, and the Egger test for publication bias. A total of eleven studies with 15692 individuals met the eligibility criteria. The prevalence of dry eye during the COVID-19 pandemic was 61.0% (95%CI: 51.8%-70.2%) globally and 56.7% (95%CI: 45.3%-68.1%) in Asia. The prevalence of dry eye had significant differences in sex and visual display time, with higher prevalence among females and visual display time of more than 4 hours per day. Subgroup analysis was performed based on diagnostic tools, study population, and average age. A significant difference was found in diagnostic tools, but no significant change in heterogeneity (P<0.05). The leave-one-out method showed stable results, and the Egger test identified no significant publication bias. The prevalence of dry eye during the COVID-19 pandemic is significantly higher than before, and a higher prevalence is found among females and those having a visual display time of more than 4 hours per day.


The early toad gets the worm: cane toads at an invasion front benefit from higher prey availability.
In biological invasions, rates of range expansion tend to accelerate through time. What kind of benefits to more rapidly dispersing organisms might impose natural selection for faster rates of dispersal, and hence the evolution of range-edge acceleration? We can answer that question by comparing fitness-relevant ecological traits of individuals at the invasion front compared with conspecifics in the same area a few years post-invasion. In tropical Australia, the rate of invasion by cane toads (Rhinella marina) has increased substantially over recent decades, due to shifts in heritable traits. Our data on field-collected cane toads at a recently invaded site in the Australian wet-dry tropics span a 5-year period beginning with toad arrival. Compared with conspecifics that we monitored in the same sites post-invasion, toads in the invasion vanguard exhibited higher feeding rates, larger energy stores, better body condition and faster growth. Three processes may have contributed to this pattern: (i) higher prey availability at the front (perhaps due to reduced competition from conspecifics); (ii) the lack of viability-reducing parasites and pathogens in invasion-front toads; and (iii) distinctive (active, fast-growing) phenotypes of the invasion-front toads. Nutritional benefits to individuals in the invasion vanguard (whether because of higher prey availability, or lower pathogen levels) thus may have conferred a selective advantage to accelerated dispersal in this system.


Interplay between the plasma membrane and cell-cell adhesion maintains epithelial identity for correct polarised cell divisions.
Polarised epithelial cell divisions represent a fundamental mechanism for tissue maintenance and morphogenesis. Morphological and mechanical changes in the plasma membrane influence the organisation and crosstalk of microtubules and actin at the cell cortex, thereby regulating the mitotic spindle machinery and chromosome segregation. Yet, the precise mechanisms linking plasma membrane remodelling to cell polarity and cortical cytoskeleton dynamics to ensure accurate execution of mitosis in mammalian epithelial cells remain poorly understood. Here, we manipulated the density of mammary epithelial cells in culture, which led to several mitotic defects. Perturbation of cell-cell adhesion formation impairs the dynamics of the plasma membrane, affecting the shape and size of mitotic cells and resulting in defects in mitotic progression and the generation of daughter cells with aberrant architecture. In these conditions, F- actin-astral microtubule crosstalk is impaired, leading to mitotic spindle misassembly and misorientation, which in turn contributes to chromosome mis-segregation. Mechanistically, we identify S100 Ca2+-binding protein A11 (S100A11) as a key membrane-associated regulator that forms a complex with E-cadherin (CDH1) and the leucine-glycine-asparagine repeat protein LGN (also known as GPSM2) to coordinate plasma membrane remodelling with E-cadherin-mediated cell adhesion and LGN-dependent mitotic spindle machinery. Thus, plasma membrane-mediated maintenance of mammalian epithelial cell identity is crucial for correct execution of polarised cell divisions, genome maintenance and safeguarding tissue integrity.


Properties and Applications of Polyvinyl Alcohol, Halloysite Nanotubes and Their Nanocomposites.
The aim of this review was to analyze/investigate the synthesis, properties, and applications of polyvinyl alcohol-halloysite nanotubes (PVA-HNT), and their nanocomposites. Different polymers with versatile properties are attractive because of their introduction and potential uses in many fields. Synthetic polymers, such as PVA, natural polymers like alginate, starch, chitosan, or any material with these components have prominent status as important and degradable materials with biocompatibility properties. These materials have been developed in the 1980s and are remarkable because of their recyclability and consideration of the natural continuation of their physical and chemical properties. The fabrication of PVA-HNT nanocomposites can be a potential way to address some of PVA's limitations. Such nanocomposites have excellent mechanical properties and thermal stability. PVA-HNT nanocomposites have been reported earlier, but without proper HNT individualization and PVA modifications. The properties of PVA-HNT for medicinal and biomedical use are attracting an increasing amount of attention for medical applications, such as wound dressings, drug delivery, targeted-tissue transportation systems, and soft biomaterial implants. The demand for alternative polymeric medical devices has also increased substantially around the world. This paper reviews individualized HNT addition along with crosslinking of PVA for various biomedical applications that have been previously reported in literature, thereby showing the attainability, modification of characteristics, and goals underlying the blending process with PVA.


Reports of substance abuse prevention programming available in schools.
Evaluations of school-based substance abuse prevention programs with schools or school districts randomly assigned to either the treatment or control condition have demonstrated effective strategies over the past 30 years. Although control schools were never considered "pure" (i.e., no other interventions were being offered), school-based programming in the 1980s did not include evidence-based interventions. Since the late 1990s, funding agencies have required schools either to select programming from approved lists of prevention strategies or to demonstrate the efficacy of the strategies that would be used. This has increased the number of schools delivering evidence-based programs to their students. As a result, "treatment as usual" is more challenging to researchers. This paper describes exposure to prevention programming as reported by 204 school administrators from 83 districts and their 19,200 students who are participating in the Adolescent Substance Abuse Prevention Study, a national randomized evaluation trial of the program, Take Charge of Your Life. In order to determine the extent of student exposure to prevention programming in both the control and treatment schools, data were collected in each of the 5 years of the study from two sources: principals and prevention coordinators and from students. The data provided by the principals and prevention coordinators indicate that the vast majority of schools assigned to the control condition offered students drug prevention programming. This finding has implications for the evaluation of Take Charge of Your Life but also for other evaluation studies. The students were asked questions regarding participation in drug education posed on annual surveys. When their responses were compared to the reports from their school principals and prevention coordinators, it was found that the students underreported exposure to drug education. A follow-up qualitative study of a sample of students suggests the need for rewording of the questions for students in future studies. The implications of our findings for evaluation studies are discussed.


Assessing contribution of bottled water in nutrient absorption using the bottled water nutritional quality index (BWNQI) in Iran.
In this study, the contribution of bottled water in the absorption of nutritional minerals in Iran has been investigated. To calculate the nutritional quality index of bottled water (BWNQI) and evaluate the contribution of bottled water in nutrient absorption; the concentration of nutrient minerals, the standard level of these elements in bottled water, the recommended amount of nutrient mineral and the total consumption of drinking water in different age-sex groups were analyzed. The results showed that the average contribution of bottled water in absorbing the recommended amount of the nutrients of fluoride (F), magnesium (Mg), calcium (Ca), sodium (Na), copper (Cu), zinc (Zn) and manganese (Mn) was 12.16, 4.98, 4.85, 2.12, 0.49, 0.33 and 0.02%, respectively. According to the BWNQI index, the bottled water quality was as follows: 53.5% poor, 36.6% marginal, 7% fair, 2.81% good. Although most of the bottled water studied in this research were mineral water, a significant portion of them had poor nutritional quality, so the addition of minerals needed by the body through bottled water should be given more attention by the bottled water manufacturers and suppliers.


Histopathology and bacteriology of postinfusion phlebitis.
Veins from patients with postinfusion phlebitis showed three types of histopathologic changes--mild, moderate and severe. Severe change was usually associated with thrombus formation. Bacterial culture of tips of devices used in infusion yielded only 10% of positive cultures. This supports the view that postinfusion phlebitis is probably due to an irritating foreign body type reaction rather than infection.


Supply and Perceived Demand for Teleophthalmology in Triage and Consultations in California Emergency Departments.
Determining the perceived supply and potential demand for teleophthalmology in emergency departments could help mitigate coverage gaps in emergency ophthalmic care. To evaluate the perceived current need for and availability of ophthalmologist coverage in California emergency departments and the potential effect of telemedicine for ophthalmology triage and consultation. Surveys were remotely administered to 187 of the 254 emergency departments throughout California via the telephone and Internet from June 30 to September 23, 2014. Emergency department nurse managers and physicians from all emergency departments listed in the California Office of Statewide Health Planning and Development database were individually surveyed to assess facility characteristics and resources as well as the perceived usefulness of teleophthalmology consultation. Data analysis was conducted from June 30, 2014, to March 11, 2015. Perceived availability of ophthalmology consultation coverage and perceived effect of telemedicine ophthalmology consultation at each facility. Of the 187 emergency departments surveyed, 18 of 37 rural facilities (48.6%) reported availability of emergency ophthalmology coverage, compared with 112 of 150 nonrural facilities (74.7%). Rural facilities reported a mean (SD) of 23.72 (14.15) miles between the facility and referral location, while nonrural facilities reported a mean of 4.41 (10.23) miles (19.3% difference). On a scale of 1 to 5 (where 1 signifies very low value and 5 signifies very high value), 124 of 187 nurse managers (66.3%) and 80 of 121 physicians (66.1%) rated teleophthalmology as having high or very high value for triage purposes. The most frequently cited potential advantage of emergency teleophthalmology was assistance in patient triage and immediate real-time electronic communication, and the most frequently cited potential disadvantages were unknown cost of contracting and maintenance and concern that eye trauma might make photographs or videos less conclusive. Availability of ophthalmology coverage for emergency eye care is limited, particularly among rural emergency departments in California. Surveyed emergency department nurse managers and physicians indicated moderately high interest and perceived value for a teleophthalmology solution for remote triage and consultation. Overall, the study suggests that teleophthalmology could play a role in mitigating coverage gaps in emergency ophthalmic care and could be further investigated through similar studies in other regions.


Synergistically interacting dopamine D1 and NMDA receptors mediate nonvesicular transporter-dependent GABA release from rat striatal medium spiny neurons.
Given the complex interactions between dopamine D1 and glutamate NMDA receptors in the striatum, we investigated the role of these receptors in transporter-mediated GABA release from cultured medium spiny neurons of rat striatum. Like NMDA receptor-mediated [(3)H]-GABA release, that induced by prolonged (20 min) dopamine D1 receptor activation was enhanced on omission of external calcium, was action potential-independent (tetrodotoxin-insensitive), and was diminished by the GABA transporter blocker nipecotic acid, indicating the involvement of transporter-mediated release. Interestingly, lowering the external sodium concentration only reduced the stimulatory effect of NMDA. Blockade of Na(+)/K(+)-ATPase by ouabain enhanced NMDA-induced but abolished dopamine-induced release. Moreover, dopamine appeared to potentiate the effect of NMDA on [(3)H]-GABA release. These effects of dopamine were mimicked by forskolin. mu-Opioid receptor-mediated inhibition of adenylyl cyclase by morphine reduced dopamine- and NMDA-induced release. These results confirm previous studies indicating that NMDA receptor activation causes a slow action potential-independent efflux of GABA by reversal of the sodium-dependent GABA transporter on sodium entry through the NMDA receptor channel. Moreover, our data indicate that activation of G-protein-coupled dopamine D1 receptors also induces a transporter-mediated increase in spontaneous GABA release, but through a different mechanism of action, i.e., through cAMP-dependent inhibition of Na(+)/K(+)-ATPase, inducing accumulation of intracellular sodium, reversal of the GABA carrier, and potentiation of NMDA-induced release. These receptor interactions may play a crucial role in the behavioral activating effects of psychostimulant drugs.


Dosage effects on heritability and maternal effects in diploid and triploid Chinook salmon (Oncorhynchus tshawytscha).
Induced triploidy (3N) in salmon results from a blockage of maternal meiosis II, and hence provides a unique opportunity to study dosage effects on phenotypic variance. Chinook salmon families were bred using a paternal half-sib breeding design (62 females and 31 males) and half of each resulting family was treated to induce triploidy. The paired families were used to test for dosage effects (resulting from triploidy) on (1) the distribution and magnitude of phenotypic variation, (2) narrow-sense heritability and (3) maternal effects in fitness-related traits (i.e., survival, size-at-age, relative growth rate and serum lysozyme activity). Quantitative genetic analyses were performed separately for diploid and triploid family groups. Triploidization resulted in significantly higher levels of phenotypic variance and substantial differences in patterns of variance distribution for growth and survival-related traits, although the patterns were reversed for lysozyme activity. Triploids exhibited higher narrow sense heritability values relative to diploid Chinook salmon. However, maternal effects estimates were generally lower in triploids than in diploids. Thus, the dosage effects resulting from adding an extra set of chromosomes to the Chinook salmon genome are primarily additive. Somewhat counterintuitively, however, the relative magnitude of the combined effects of dominance, epistasis and maternal effects is not affected by dosage. Our results indicate that inheritance of fitness-related quantitative traits is profoundly affected by dosage effects associated with induced triploidy, and that triploidization can result in unpredictable performance and fitness outcomes.


In Spinal Anaesthesia for Cesarean Section the Temperature of Bupivacaine Affects the Onset of Shivering but Not the Incidence: A Randomized Control Trial.
Postoperative shivering is a frequent event after cesarean section under spinal anaesthesia. Shivering is uncomfortable for the patient and may interfere with monitoring. The exact aetiology of shivering is unknown and therefore has no definite treatment. The temperature of injectate affects the spread of drug and so its effect. Therefore the aim of this study was to compare the effect of temperature of bupivacaine on post-spinal shivering in cesarean section. In this prospective, randomized, controlled, double-blind clinical trial 105 ASA-I/II pregnant women scheduled for caesarean section under spinal anaesthesia were selected and randomized into three groups of 35 each. In all pregnant women spinal anaesthesia was achieved with 2.2 ml of 0.5% hyperbaric bupivacaine given either at L3-L4 or L4-L5 interspace. The temperature of bupivacaine was adjusted to 4°C (group T4), 22°C (group T22) and 37°C (group T37). Shivering characteristic, onset and incidence was noted. All three groups were compared using analysis of variance (ANOVA), adverse effects was compared using chi-square test and Kruskal-Wallis H-test. The p-value < 0.05-considered as significant and p-value <0.01-considered highly significant. There were no differences between the groups regarding age, weight, height, amount of fluid used and blood loss. The incidence of shivering was 51.42%, 51.42% and 45.71% in group T4, group T22 and group T37 respectively, this difference in the incidence was statistically not significant (p=0.858). However, the onset of shivering was earliest (9.87±1.82 min) in group T4 as compared to 14.27±3.02 min and 12.16±2.89 min in group T22 and group T37 respectively and this difference in the onset was highly significant (p= 0.0001). In spinal anaesthesia for cesarean section, the temperature of bupivacaine does not influence the overall incidence of post spinal shivering; however cold bupivacaine can provoke early onset of shivering.


Lipid-lowering drugs: are adverse effects predictable and reversible?
Most of the currently available statins and fibric acid derivatives have been implicated in causing complications either in monotherapy or in combination therapy. Adverse events occur more often with the statins that are metabolized via the CYP enzyme system and its 3A4, 2C9 or 2C19 paths. All compounds interfering with the same cytochrome system may either impair or enhance the elimination of statins. Other factors predisposing to adverse effects are age, female sex, renal insufficiency, electrolyte disturbances, infections and trauma. Complications chiefly concern the hepatic function, skeletal muscles and peripheral nerves. The major adverse effect is myopathy, up to rhabdomyolysis with ensuing acute renal insufficiency. Fibrates bind to peroxisome proliferator-activated nuclear receptors alpha, with subsequent stimulation of fatty acid oxidation and reduction in the rate of hepatic lipid generation. Fibrates are associated with a number of adverse effects, including liver enzyme elevations, gastrointestinal side effects and rhabdomyolysis. The combination of statins with fibrates may cause serious complications and should be avoided when possible. In order to prevent or minimize adverse clinical outcomes, patients should be closely monitored and informed of the most common symptoms.


Myoglobin expression in prostate cancer is correlated to androgen receptor expression and markers of tumor hypoxia.
Recent studies identified unexpected expression and transcriptional complexity of the hemoprotein myoglobin (MB) in human breast cancer but its role in prostate cancer is still unclear. Expression of MB was immunohistochemically analyzed in three independent cohorts of radical prostatectomy specimens (n = 409, n = 625, and n = 237). MB expression data were correlated with clinicopathological parameters and molecular parameters of androgen and hypoxia signaling. Expression levels of novel tumor-associated MB transcript variants and the VEGF gene as a hypoxia marker were analyzed using qRT-PCR. Fifty-three percent of the prostate cancer cases were MB positive and significantly correlated with androgen receptor (AR) expression (p < 0.001). The positive correlation with CAIX (p < 0.001) and FASN (p = 0.008) as well as the paralleled increased expression of the tumor-associated MB transcript variants and VEGF suggest that hypoxia participates in MB expression regulation. Analogous to breast cancer, MB expression in prostate cancer is associated with steroid hormone signaling and markers of hypoxia. Further studies must elucidate the novel functional roles of MB in human carcinomas, which probably extend beyond its classic intramuscular function in oxygen storage.


Mesenchymal stromal cells in renal ischemia/reperfusion injury.
Ischemia/reperfusion (I/R) injury is an inevitable consequence of organ transplantation and a major determinant of patient and graft survival in kidney transplantation. Renal I/R injury can lead to fibrosis and graft failure. Although the exact sequence of events in the pathophysiology of I/R injury remains unknown, the role of inflammation has become increasingly clear. In this perspective, mesenchymal stromal cells (MSCs) are under extensive investigation as potential therapy for I/R injury, since MSCs are able to exert immune regulatory and reparative effects. Various preclinical studies indicate the beneficial effects of MSCs in ameliorating renal injury and accelerating tissue repair. These versatile cells have been shown to migrate to sites of injury and to enhance repair by paracrine mechanisms instead of by differentiating and replacing the injured cells. The first phase I studies of MSCs in human renal I/R injury and kidney transplantation have been started, and results are awaited soon. In this review, preliminary results and opportunities of MSCs in human renal I/R injury are summarized. We might be heading towards a cell-based paradigm shift in the treatment of renal I/R injury.


Lipopolysaccharide chemotypes of Burkholderia cepacia.
Burkholderia cepacia is an important pathogen in patients with cystic fibrosis (CF) and much is now known of its epidemiology. In contrast, its virulence mechanisms are poorly understood. The lipopolysaccharide (LPS) of B. cepacia, a well-recognised virulence factor of other gram-negative bacteria, is known to be strongly endotoxic in vitro. The aim of this study was to observe if there were any links between the structure of B. cepacia LPS and virulence. This has been investigated by polyacrylamide gel electrophoresis and immunoblotting to define the chemotype and antigenic cross reactivity of B. cepacia LPS. Strains (16) belonging to different genomovars of the B. cepacia complex were selected to represent epidemic and non-epidemic clinical isolates and environmental strains. All strains belonging to genomovars I and II (the latter now renamed B. multivorans) had smooth LPS. However, isolates belonging to genomovar III, the group to which most of the epidemic CF isolates belong - including the highly transmissible strain (ET 12) which has been found in both the UK and North America - were of either rough or smooth LPS chemotype. In this study, B. cepacia J2315 represents the ET 12 lineage, and has a rough chemotype. Rabbit antiserum raised to strain J2315 revealed that the LPS core of this strain was antigenically related to some but not all other genomovar III strains, but it also cross-reacted strongly with all B. multivorans (genomovar II) and most genomovar I strains. Intra-strain phenotypic variation was demonstrated between bacteria grown in broth or on solid agar with a concomitant variation in antigenic cross reactivity. There was no clear evidence to associate any particular LPS phenotype with epidemic or non-epidemic strains, but changes in phenotype in vitro may provide clues to the survival and adaptability of B. cepacia in hostile environments and possibly to its ability to produce an inflammatory response in vivo.


Devolution to democratic health authorities in Saskatchewan: an interim report.
In 1995 Saskatchewan adopted a district health board structure in which two-thirds of members are elected and the rest are appointed. This study examines the opinions of board members about health care reform and devolution of authority from the province to the health districts. All 357 members of Saskatchewan district health boards were surveyed in 1997; 275 (77%) responded. Analyses included comparisons between elected and appointed members and between members with experience as health care providers and those without such experience, as well as comparisons with hypotheses about how devolution would develop, which were advanced in a 1997 report by another group. Most respondents felt that devolution had resulted in increased local control and better quality of decisions. Ninety-two percent of respondents believed extensive reforms were necessary and 83% that changes made in the previous 5 years had been for the best. However, 56% agreed that there was no clear vision of the reformed system. A small majority (59%) perceived health care reform as having been designed to improve health rather than reduce spending, contrary to a previous hypothesis. Many respondents (76%) thought that boards were legally responsible for things over which they had insufficient control, and 63% perceived that they were too restricted by rules laid down by the provincial government, findings that confirm the expectation of tensions surrounding the division of authority. Respondents with current or former experience as health care providers were less likely than nonprovider respondents to believe that nonphysician health care providers support decisions made by the regional health boards (45% v. 63%, p = 0.02), a result that confirmed the contention that the role of health care providers on the boards would be a source of tension. Members of Saskatchewan district health boards supported the general goals of health care reform and believed that changes already undertaken had been positive. There were few major differences in views between appointed and elected members and between provider and nonprovider members. However, tensions related to authority and representation will require resolution.


A rare case of silent transmural myocardial infarction with diffuse ST elevations complicated by concomitant severe hyperkalemia.
It is well described that certain group of patients do not display the typical symptoms of myocardial infarction (MI). Elderly patients, diabetics and those with previous coronary artery bypass graft surgery are at high risk for silent MI. The diagnosis of Acute MI in the emergency room (ER) is mainly based on the electrocardiogram (EKG) findings of ST elevations or new onset left bundle branch block which is supported by the clinical presentation and positive biomarkers when present. The diagnoses can sometimes become challenging when the patient is asymptomatic and has coincidental finding of hyperkalemia with diffuse ST segment elevations simulating that seen with electrolyte disturbance. Despite the well known pseudoinfarction pattern of hyperkalemia, acute MI should be ruled out first. A high index of suspicion is needed, especially in high risk patients. We think that in rare clinical situation when the diagnosis is in doubt, MI should be ruled out, as time has a high impact on patient mortality. An urgent bedside echocardiogram is very beneficial in excluding regional wall motion abnormalities and preventing any delay in destination therapy for transmural MI. We present a 67 years old female with history of diabetes and chronic kidney disease sent by her nephrologist to the ER for severe hyperkalemia (Potassium 7.2 milliequivalent/L). She was found to have ST elevations on EKG despite having no chest pain or distress. On cardiac catheterization she had a total occlusion of the proximal left circumflex artery, with a filling defect consistent with large thrombus.


A proposal for a nonreinstatement rule in unfair labor practice cases involving patient abuse.
Nursing home discharges of employees based on patient abuse raise a difficult issue when the motivating factor for the disciplinary action is union activism. A tension is created between the rights of employees to engage in protected concerted activity and the rights of patients to quality care. In 1974, Congress passed the Health Care Institutions Amendments, which granted to non-profit health care workers collective organizing and bargaining rights substantially similar to those which workers in other industries had enjoyed for decades under the National Labor Relations Act. Congress intended to give health care workers only that degree of parity, however, which is compatible with the provision of high quality patient care. The agency charged with enforcing the Act, the National Labor Relations Board (NLRB), has failed to distinguish employee misconduct in industrial settings from patient abuse in health care institutions when fashioning remedies for discriminatorily discharged union activists. The NLRB typically has ordered the reinstatement, with back pay, of the patient abuser as the patient's primary care-giver. This Article suggests that a front pay remedy is more appropriate to these cases because it protects the patient's right to be free from abuse without sacrificing employee unionization rights.


Diet quality as assessed by the Healthy Eating Index, the Alternate Healthy Eating Index, the Dietary Approaches to Stop Hypertension score, and health outcomes: a systematic review and meta-analysis of cohort studies.
Dietary patterns consider synergistic effects compared with isolated foods or nutrients on health outcomes. The aim of this systematic review and meta-analysis was to examine the associations of diet quality as assessed by the Healthy Eating Index (HEI), the Alternate Healthy Eating Index (AHEI), and the Dietary Approaches to Stop Hypertension (DASH) score and the risk of all-cause mortality, cardiovascular mortality or incidence, cancer mortality or incidence, type 2 diabetes mellitus, and neurodegenerative diseases. A literature search was performed using the electronic databases MEDLINE, SCOPUS, and EMBASE with an end date of May 10, 2014. Study-specific risk ratios were pooled using a random effect model by the Cochrane software package Review Manager 5.2. Fifteen cohort studies (34 reports), including 1,020,642 subjects, met the criteria and were included in the meta-analysis. Diets of the highest quality, as assessed by the HEI, AHEI, and DASH score, resulted in a significant risk reduction (RR) for all-cause mortality (RR 0.78, 95% CI 0.76 to 0.80; P<0.00001; I²=61%, 95% CI 20% to 81%), cardiovascular disease (incidence or mortality) (RR 0.78, 95% CI 0.75 to 0.81; P<0.00001; I²=45%, 95% CI 13% to 66%), cancer (incidence or mortality) (RR 0.85, 95% CI 0.82 to 0.88; P<0.00001; I²=77%, 95% CI 68% to 84%), and type 2 diabetes mellitus (RR 0.78, 95% CI 0.72 to 0.85; P<0.00001; I²=74%, 95% CI 52% to 86%). Differences observed for neurodegenerative diseases were not significant. Egger regression tests provided no evidence of publication bias. Diets that score highly on the HEI, AHEI, and DASH are associated with a significant reduction in the risk of all-cause mortality, cardiovascular disease, cancer, and type 2 diabetes mellitus by 22%, 22%, 15%, and 22%, respectively, and therefore is of high public health relevance.


[Immune system and alcoholic liver disease].
It is well established that alcoholism is associated with imbalanced immune responses. To date, most relevant finding reported is the existence of an immunodepressed state which leads to a higher risk of suffering from severe infections in alcoholic patients. However, recent studies have shown that ethanol intake is followed by changes involving the synthesis and serum levels of specific cytokines as well as the activation of several different subsets of cytotoxic lymphocytes, that could be involved in the development of alcoholic liver disease. Accordingly, tumor necrosis factor-alpha plays a key role in the development of alcoholic liver damage through the induction of both apoptosis and necrosis of hepatocytes. This cytokine, together with interleukin (IL) 1, IL6 and several chemokines, facilitate the development of inflammation of the liver. Additionally, both transforming growth factor-beta and platelet-derived growth factor, act over stellate cells favouring hepatic fibrogenesis. The advances in the knowledge of the immunological mechanisms involved in alcoholic liver disease may lead to the discovery of new potential therapeutic targets, which may modify disease outcome in the near future.


Impaired TrkB Signaling Underlies Reduced BDNF-Mediated Trophic Support of Striatal Neurons in the R6/2 Mouse Model of Huntington's Disease.
The principal projection neurons of the striatum are critically dependent on an afferent supply of brain derived neurotrophic factor (BDNF) for neurotrophic support. These neurons express TrkB, the cognate receptor for BDNF, which activates signaling pathways associated with neuronal survival and phenotypic maintenance. Impairment of the BDNF-TrkB pathway is suspected to underlie the early dysfunction and prominent degeneration of striatal neurons in Huntington disease (HD). Some studies in HD models indicate that BDNF supply is reduced, while others suggest that TrkB signaling is impaired earlier in disease progression. It remains important to determine whether a primary defect in TrkB signaling underlies reduced neurotrophic support and the early vulnerability of striatal neurons in HD. Using the transgenic R6/2 mouse model of HD we found that prior to striatal degeneration there are early deficits in striatal protein levels of activated phospho-TrkB and the downstream-regulated protein DARPP-32. In contrast, total-TrkB and BDNF protein levels remained normal. Primary neurons cultured from R6/2 striatum exhibited reduced survival in response to exogenous BDNF applications. Moreover, BDNF activation of phospho-TrkB and downstream signal transduction was attenuated in R6/2 striatal cultures. These results suggest that neurotrophic support of striatal neurons is attenuated early in disease progression due to defects in TrkB signal transduction in the R6/2 model of HD.


How Can Single Sensory Neurons Predict Behavior?
Single sensory neurons can be surprisingly predictive of behavior in discrimination tasks. We propose this is possible because sensory information extracted from neural populations is severely restricted, either by near-optimal decoding of a population with information-limiting correlations or by suboptimal decoding that is blind to correlations. These have different consequences for choice correlations, the correlations between neural responses and behavioral choices. In the vestibular and cerebellar nuclei and the dorsal medial superior temporal area, we found that choice correlations during heading discrimination are consistent with near-optimal decoding of neuronal responses corrupted by information-limiting correlations. In the ventral intraparietal area, the choice correlations are also consistent with the presence of information-limiting correlations, but this area does not appear to influence behavior, although the choice correlations are particularly large. These findings demonstrate how choice correlations can be used to assess the efficiency of the downstream readout and detect the presence of information-limiting correlations.


Lack of TRPV1 inhibits cystitis-induced increased mechanical sensitivity in mice.
Transient receptor potential vanilloid 1 (TRPV1) is highly expressed in primary afferent neurons. Tissue damage generates an array of chemical mediators that activate and sensitize afferent nerve fibers, and sensitization of afferent nerve fibers plays an important role in development of visceral pain. We investigated participation of TRPV1 in visceral pain associated with bladder inflammation induced in mice by systemic treatment with cyclophosphamide or intravesical instillation of acrolein. The effects of experimental cystitis on bladder function (an indicator of visceral pain) and the threshold of response to mechanical or thermal stimuli of the hind paws were investigated using TRPV1 knock-out (KO) and congenic wild-type (WT) mice. We found that cystitis induced bladder mechanical hyperreactivity and increased mechanical sensitivity of hind paws in WT, but not in TRPV1 KO mice. Lack of functional TRPV1 did not inhibit development of histological evidence of bladder inflammation, or increased expression of mRNAs for nerve growth factor, endothelial nitric oxide synthase, cyclooxygenase-2 and bradykinin receptors in urothelium. Cystitis did not affect the threshold of response to thermal stimuli in WT or KO mice. These results suggest that TRPV1 is essential for cystitis-induced bladder mechanical hyperreactivity. Also, TRPV1 participates in development of visceral pain, as reflected by referred increased mechanosensitivity in peripheral tissues in the presence of visceral inflammation.


Cerebrovascular Physiology During Pediatric Extracorporeal Membrane Oxygenation: A Multicenter Study Using Transcranial Doppler Ultrasonography.
To explore changes to expected, age-related transcranial Doppler ultrasound variables during pediatric extracorporeal membrane oxygenation. Prospective, observational, multicenter study. Tertiary care PICUs. Children 1 day to 18 years old requiring veno arterial extracorporeal membrane oxygenation. Participants underwent daily transcranial Doppler ultrasound measurement of bilateral middle cerebral artery flow velocities. Acute neurologic injury was diagnosed if seizures, cerebral hemorrhage, or diffuse cerebral ischemia was detected. Fifty-two children were enrolled and analyzed. In the 44 children without acute neurologic injury, there was a significant reduction in systolic flow velocity and mean flow velocity compared with predicted values over time (F [8, 434] = 60.44; p ≤ 0.0001, and F [8, 434] = 17.61; p ≤ 0.0001). Middle cerebral artery systolic flow velocity was lower than predicted on extracorporeal membrane oxygenation days 1-5, and mean flow velocity was lower than predicted on extracorporeal membrane oxygenation days 1-3. In the six infants less than 90 days old suffering diffuse cerebral ischemia, middle cerebral artery systolic flow velocity, mean flow velocity, and diastolic flow velocity from extracorporeal membrane oxygenation days 1-9 were not significantly different when compared with children of similar age in the cohort that did not suffer acute neurologic injury (systolic flow velocity F [8, 52] = 0.6659; p = 0.07 and diastolic flow velocity F [8, 52] = 1.4; p = 0.21 and mean flow velocity F [8, 52] = 1.93; p = 0.07). Pulsatility index was higher in these infants over time than children of similar age in the cohort on extracorporeal membrane oxygenation that did not suffer acute neurologic injury (F [8, 52] = 3.1; p = 0.006). No patient in the study experienced cerebral hemorrhage. Flow velocities in the middle cerebral arteries of children requiring extracorporeal membrane oxygenation are significantly lower than published normative values for critically ill, mechanically ventilated, sedated children. Significant differences in measured systolic flow velocity, diastolic flow velocity, and mean flow velocity were not identified in children suffering ischemic injury compared with those who did not. However, increased pulsatility index may be a marker for ischemic injury in young infants on extracorporeal membrane oxygenation.


Management of a swallowed denture: our experience with 34 patients.
Background: Swallowed dentures can present with upper aerodigestive tract obstruction needing urgent intervention. Removing such an ingested denture can prove challenging and needs careful planning. Aim: To share our experience of managing patients with a swallowed denture focusing on the practical aspects of denture removal along with relevant literature review. We aim to raise a public health message on the safety aspect of usage of dentures. Subjects and methods: A retrospective analysis of the patients managed by our team in the ENT department at two hospitals in Scotland, over 10 years (2009-2019), who were found to have swallowed a denture. Data on demographics, clinical history, examination findings and management of patients were collected and analysed using Microsoft Excel. Results: A total of 34 patients were admitted with a swallowed denture, of which 24 (71%) were male and 10 (29%) were female. The mean age was 60 years (range 17-83). Of the 34 patients, 2 had a feeling of something stuck in the throat but were able to eat and drink; the rest of the patients complained about dysphagia and pain in the throat, with 2 patients also showing signs of respiratory distress. Twenty-four (71%) patients required denture removal under general anaesthetic in the theatre; 20 (59%) by rigid oesophagoscopy, 1 with tracheostomy (3%), 1 with (3%) laparoscopy and gastrostomy, and 2 (6%) with external neck exploration. Seven (20%) patients were taken to the theatre and the denture was removed with Magill forceps under light sedation using intubating laryngoscope or video laryngoscope. In 1 patient (3%), the denture material was successfully removed under flexible pharyngolaryngoscopy guidance in the clinic without sedation. The final 2 (6%) patients were reassured as no foreign body was seen on flexible laryngoscopy. Conclusion: In the absence of a clear evidence of denture ingestion, a detailed history and examination are needed to identify this serious pathology. Once confirmed, the ingested denture should be removed as soon as possible to minimize the risk of serious complications.


THE EFFECTS OF ANTIMONY UPTAKE ON THE LOCATION AND PAIRING OF SCHISTOSOMA MANSONI.
The rate of uptake and elimination of antimony by both male and female schistosomes has been measured. It has been confirmed that the female schistosome is more susceptible to antimony therapy than the male since she absorbs more drug. The changes in location of the worms and the separation of the sexes following one or more injections of antimony potassium tartrate have been related to the different rates of uptake and elimination of the drug by the two sexes. The suppressive treatment of schistosomiasis is discussed in the light of these observations.


Effects of tocopherol (vitamin E) acid succinate on morphological alterations and growth inhibition in melanoma cells in culture.
The effects of various forms of tocopherol (vitamin E) on the growth and differentiation of mouse melanoma (B-16) and mouse fibroblast (L-cells) cells in culture were studied. D-alpha-tocopherol acid succinate induced morphological alterations and growth inhibition in melanoma cells. When vitamin E acid succinate was removed 4 days after treatment, the above changes remained irreversible for a period of 24 hr, after which resistant cells and partially affected cells renewed cell division and eventually reached confluency. The relative efficacy of D and DL forms of vitamin E acid succinate remains to be evaluated. However, other forms of vitamin E such as DL-alpha-tocopherol free alcohol, Aquasol DL-alpha-tocopherol acetate, DL-alpha-tocopherol nicotinate, or sodium succinate with an equivalent volume of ethanol, at similar concentrations, were ineffective. Vitamin E acid succinate at similar concentrations did not induce morphological changes in fibroblasts. Melanoma cells were about 2-fold more sensitive to vitamin E acid succinate than were fibroblasts for the criterion of growth inhibition. Vitamin E acid succinate-induced morphological changes and growth inhibition in melanoma cells were expressed in hormone-supplemented serum-free medium, but the concentration requirement was about 5 times less than that needed in serum-supplemented medium. Although cyclic adenosine 3': 5'-monophosphate-stimulating agents are known to cause growth inhibition and morphological changes in melanoma cells in culture, vitamin E acid succinate-induced morphological alterations in melanoma cells are no mediated by a rise in cellular cyclic adenosine 3':5'-monophosphate. Ethanol was sufficient to increase the melanin content in melanoma cells. These data show that vitamin E acid succinate may be a potentially useful tumor therapeutic agent.


The role of the adenosinergic pathway in immunosuppression mediated by human regulatory T cells (Treg).
Tumor-induced dysfunction of immune cells is a common problem in cancer. Tumors induce immune suppression by many different mechanisms, including accumulation of regulatory T cells (Treg). Adaptive Treg (Tr1) generated in the tumor microenvironment express CD39 and CD73 ectonucleotidases, produce adenosine and are COX2+PGE2+. Adenosine and PGE2 produced by Tr1 or tumor cells bind to their respective receptors on the surface of T effector cells (Teff) and cooperate in up-regulating cytosolic 3'5'-cAMP levels utilizing adenylyl cyclase isoform 7 (AC-7). In Teff, increased cAMP mediates suppression of anti-tumor functions. Treg, in contrast to Teff, seem to require high cAMP levels for mediating suppression. This differential requirement of Treg and Teff for cAMP offers an opportunity for pharmacologic interventions using selected inhibitors of the adenosine/PGE2 pathways. Blocking of adenosine/PGE2 production by Tr1 or blocking binding of these factors to their receptors on T cells or inhibition of cAMP synthesis in Teff all represent novel therapeutic strategies that used in combination with conventional therapies could restore anti-tumor functions of Teff . At the same time, these inhibitors could disarm Tr1 cells by depriving them of the factors promoting their generation and activity or by down-regulating 3'5'-cAMP levels. Thus, the pharmacologic control of Treg-Teff interactions offers a novel strategy for restoration of anti-tumor Teff functions and silencing of Treg. Used in conjunction with anti-cancer drugs or with immune therapies, this strategy has a potential to improve therapeutic effects by preventing or reversing tumor-induced immune suppression.


Virtual reality enhancing medical education and practice: Brief communication.
The COVID-19 pandemic has become a major cause of rapid globalization and digitization of educational institutions, including medical education. The adaptation to digital technologies is the purpose of best education and training practices in the development of the academic medical curriculum. Virtual reality (VR) is embraced by the 3D environment and network resources which allow the expansion of VR from the entertainment industry to the education industry. This brief communication explains our understanding and the challenges in adopting VR technologies for medical training at an academic medical center. Advancement in VR technology assists medical institutes to strategize for the further development of medical training and education. There is a timely need for persistence to make the VR content accessible widely and open source. There is an urgent need for collaboration of medical institutes and technology industries on the development of education-related VR content and simulations.


Difficulties accessing health care services during the COVID-19 pandemic in Canada: examining the intersectionality between immigrant status and visible minority status.
Difficulties accessing health care services can result in delaying in seeking and obtaining treatment. Although these difficulties are disproportionately experienced among vulnerable groups, we know very little about how the intersectionality of realities experienced by immigrants and visible minorities can impact their access to health care services since the pandemic. Using Statistics Canada's Crowdsourcing Data: Impacts of COVID-19 on Canadians-Experiences of Discrimination, we combine two variables (i.e., immigrant status and visible minority status) to create a new variable called visible minority immigrant status. This multiplicative approach is commonly used in intersectionality research, which allows us to explore disadvantages experienced by minorities with multiplicative identities. Main results show that, compared to white native-born, visible minority immigrants are less likely to report difficulties accessing non-emergency surgical care (OR = 0.55, p < 0.001), non-emergency diagnostic test (OR = 0.74, p < 0.01), dental care (OR = 0.71, p < 0.001), mental health care (OR = 0.77, p < 0.05), and making an appointment for rehabilitative care (OR = 0.56, p < 0.001) but more likely to report difficulties accessing emergency services/urgent care (OR = 1.46, p < 0.05). We conclude that there is a dynamic interplay of factors operating at multiple levels to shape the impact of COVID-19 related needs to be addressed through changes in social policies, which can tackle unique struggles faced by visible minority immigrants.


Finite Element Simulation of Ionic Electrodiffusion in Cellular Geometries.
Mathematical models for excitable cells are commonly based on cable theory, which considers a homogenized domain and spatially constant ionic concentrations. Although such models provide valuable insight, the effect of altered ion concentrations or detailed cell morphology on the electrical potentials cannot be captured. In this paper, we discuss an alternative approach to detailed modeling of electrodiffusion in neural tissue. The mathematical model describes the distribution and evolution of ion concentrations in a geometrically-explicit representation of the intra- and extracellular domains. As a combination of the electroneutral Kirchhoff-Nernst-Planck (KNP) model and the Extracellular-Membrane-Intracellular (EMI) framework, we refer to this model as the KNP-EMI model. Here, we introduce and numerically evaluate a new, finite element-based numerical scheme for the KNP-EMI model, capable of efficiently and flexibly handling geometries of arbitrary dimension and arbitrary polynomial degree. Moreover, we compare the electrical potentials predicted by the KNP-EMI and EMI models. Finally, we study ephaptic coupling induced in an unmyelinated axon bundle and demonstrate how the KNP-EMI framework can give new insights in this setting.


Psychophysical test for the independence of perception and action.
Research has suggested that perception and action are independent (see M. A. Goodale & A. Haffenden, 1998). The authors used the Ebbinghaus illusion to test this hypothesis in 2 experiments. Verbal reports of perceived size were compared with maximum grip aperture during grasping (Experiment 1) and manual reports of perceived size (Experiment 2). A multidimensional signal detection analysis was used to distinguish among the possible interactions between the two processes in each experiment (H. Kadlec & J. T. Townsend, 1992a, 1992b). In Experiment 1, the percepts were independent, and there were no interactions between processes for verbal and visuomotor responses. In Experiment 2, the percepts associated with verbal and manual reports were independent, but the processes interacted at the levels of the stimulus information and the decision rules used to transform each percept into a response.


Amino acid kinetics in the critically ill.
Stable isotope methods have been used for many years to assess whole body protein and amino acid kinetics in critically ill patients. In recent years, new isotope approaches and tracer insights have been developed. The tracer pulse approach has some advantages above the established primed-continuous tracer infusion approach because of the high amount of metabolic information obtained, easy applicability, and low tracer costs. Effects of disease severity and sex on amino acid kinetics in ICU patients will also be addressed. Current knowledge was synthesized on specific perturbations in amino acid metabolism in critically ill patients, employing novel methodologies such as the pulse tracer approach and computational modeling. Variations were evaluated in amino acid production and linked to severity of critical illness, as measured by SOFA score, and sex. Production of the branched-chain amino acids (BCAAs), glutamine, tau-methylhistidine and hydroxyproline were elevated in critical illness, likely related to increased transamination of the individual BCAAs or increased breakdown of proteins. Citrulline production was reduced, indicative of impaired gut mucosa function. Sex and disease severity independently influenced amino acid kinetics in ICU patients. Novel tracer and computational approaches have been developed to simultaneously measure postabsorptive kinetics of multiple amino acids that can be used in critical illness. The collective findings lay the groundwork for targeted individualized nutritional strategies in ICU settings aimed at enhancing patient outcomes taking into account disease severity and sex.


On the role of alpha 1-acid glycoprotein in lignocaine accumulation following myocardial infarction.
1 Blood plasma and free lignocaine concentrations have been measured 12 h after beginning a constant infusion of 2 mg/min and again at the end of the infusion (36-72 h) in five patients with myocardial infarction (MI) and compared with five control patients who did not develop objective evidence of MI. 2 In MI patients, total plasma concentration rose significantly between 12 h and the end of infusion. Because of an increase in alpha 1 acid glycoprotein (AAG) plasma binding increased, so that free drug concentration did not change. The rise in whole blood concentration was less than that in plasma as a result of drug redistribution out of red cells due to enhanced binding. 3 In control patients, neither blood nor plasma concentrations changed with time and plasma binding remained constant. Free drug concentrations, however, rose slightly. 4 The concentrations of GX and MEGX remained unchanged in all patients, but the ratio of lignocaine/MEGX concentrations fell in controls but rose in MI patients. 5 Pharmacokinetic modelling suggested that at least some of the rise in blood lignocaine concentration was due to reduced clearance resulting from enhanced plasma binding. 6 We conclude that the rise in AAG following MI is responsible for increased plasma binding and drug redistribution within blood. These changes, together with a reduction in lignocaine clearance, can explain much of the phenomenon of lignocaine accumulation in MI.


Stress and safety of maternal and newborn healthcare workers early in the COVID-19 pandemic: a repeat cross-sectional analysis from a global online survey from March 2020 to March 2021.
This study aims to characterise the physical and psychological well-being of maternal and newborn healthcare workers (MNHCWs) during the COVID-19 pandemic. Observational repeated cross-sectional study. An online questionnaire was distributed to MNHCWs around the globe in three separate rounds from March 2020 to March 2021. Total samples of N=1357 (round 1) and N=420 (round 3) primarily consisted of doctors, midwives and nurses in maternal and newborn specialties. Samples represented all WHO regions, with 33% (round 1) and 42% (round 3) from low- or middle-income countries (LMICs). Responses from rounds 1 (March-June 2020) and 3 (December 2020-March 2021) were analysed to measure self-reported levels of relative stress and workplace protection from COVID-19, while associated factors were determined through multivariable ordinal logistic regression. In round 1, 90% of MNHCWs reported increased stress levels and 45% reported insufficient personal protective equipment (PPE) access. Nurses and physicians were less likely to report increased stress than midwives at the pandemic onset. Factors associated with increased stress included being female, being from an LMIC and insufficient PPE. In round 3, 75% reported similar or increased stress while 10% reported insufficient PPE. In both rounds, over 50% of MNHCWs felt relatively or completely unprotected from COVID-19 in the workplace. Those from LMICs were more likely to report feeling unprotected, while receiving organisational information that valued safety was associated with better feelings of protection in the workplace. Among our international sample of MNHCWs, we observed high rates of self-reported stress increase at the start of the pandemic with persistence or increase up to a year later. High rates of feeling unprotected persisted even as PPE became more available. These results may inform interventions needed to support and protect MNHCWs during this and future pandemics.


Metabolism of benzo[a]pyrene by human melanocytes in culture.
Benzo[a]pyrene (B[a]P) is a ubiquitous environmental pollutant and known skin carcinogen. In the present study, in vitro addition of [3H]B[a]P to normal human melanocytes in culture, isolated from adult human skin, resulted in the metabolism of [3H]B[a]P both intracellularly and extracellularly. HPLC analysis showed that [3H]B[a]P-9,10- and 7,8-diol were the major intracellular and extracellular metabolites followed by 3,6-quinone, 9-hydroxy and 3-hydroxy metabolites. Significant amounts of the [3H]B[a]P metabolites were found to be present in the sonicated cell suspension and culture medium as the glucuronide and sulfate conjugates. In total 37.3% of the [3H]B[a]P added in the culture medium was metabolized by melanocytes, of which 21.1% was quantified as the intracellular and 16.2% as the extracellular metabolites. Our data show that human melanocytes are capable of metabolizing B[a]P.


Factors associated with sources, transport, and fate of chloroform and three other trihalomethanes in untreated groundwater used for drinking water.
Multiple lines of evidence for indicating factors associated with the sources, transport, and fate of chloroform and three other trihalomethanes (THMs) in untreated groundwater were revealed by evaluating low-level analytical results and logistic regression results for THMs. Samples of untreated groundwater from wells used for drinking water were collected from 1996-2007 from 2492 wells across the United States and analyzed for chloroform, bromodichloromethane, dibromochloromethane, and bromoform by a low-level analytical method implemented in April 1996. Using an assessment level of 0.02 μg/L, chloroform was detected in 36.5% of public-well samples and 17.6% of domestic-well samples, with most concentrations less than 1 μg/L. Brominated THMs occurred less frequently than chloroform but more frequently in public-well samples than domestic-well samples. For both public and domestic wells, THMs occurred most frequently in urban areas. Logistic regression analyses showed that the occurrence of THMs was related to nonpoint sources such as urban land use and to point sources like septic systems. The frequent occurrence and concentration distribution pattern of THMs, as well as their frequent co-occurrence with other organic compounds and nitrate, all known to have anthropogenic sources, and the positive associations between THM occurrence and dissolved oxygen and recharge indicate the recycling of water that contains THMs and other anthropogenic contaminants.


Lysine Demethylase LSD1 Coordinates Glycolytic and Mitochondrial Metabolism in Hepatocellular Carcinoma Cells.
The hallmark of most cancer cells is the metabolic shift from mitochondrial to glycolytic metabolism for adapting to the surrounding environment. Although epigenetic modification is intimately linked to cancer, the molecular mechanism, by which epigenetic factors regulate cancer metabolism, is poorly understood. Here, we show that lysine-specific demethylase-1 (LSD1, KDM1A) has an essential role in maintaining the metabolic shift in human hepatocellular carcinoma cells. Inhibition of LSD1 reduced glucose uptake and glycolytic activity, with a concurrent activation of mitochondrial respiration. These metabolic changes coexisted with the inactivation of the hypoxia-inducible factor HIF1α, resulting in a decreased expression of GLUT1 and glycolytic enzymes. In contrast, during LSD1 inhibition, a set of mitochondrial metabolism genes was activated with the concomitant increase of methylated histone H3 at lysine 4 in the promoter regions. Consistently, both LSD1 and GLUT1 were significantly overexpressed in carcinoma tissues. These findings demonstrate the epigenetic plasticity of cancer cell metabolism, which involves an LSD1-mediated mechanism.


How to design a hospitalist service. An academic health system case study.
Medical centers throughout the country are employing hospitalists--dedicated inpatient specialists--to care for patients during hospitalization in place of their primary care provider. There has been expansive growth of hospitalist programs as the number of practicing hospitalists has doubled in two years from approximately 1,500 to over 3,000. In July 1997, the department of medicine at University Hospitals of Cleveland, the primary teaching affiliate of Case Western Reserve University, created a hospitalist service. This paper will discuss the design of a hospitalist service at a major academic medical center, the factors that were considered during the design stage and financial feasibility.


The Novel Atypical Antipsychotic Lurasidone Affects Cytochrome P450 Expression in the Liver and Peripheral Blood Lymphocytes.
Lurasidone is a novel atypical antipsychotic drug acting on dopaminergic, serotonergic and noradrenergic receptors; it is applied for the long-term treatment of schizophrenia and depression in patients with bipolar disorders. We aimed at performing a comparative study on the influence of chronic treatment with lurasidone on the expression of cytochrome P450 enzymes in the liver and in peripheral blood lymphocytes, and to evaluate the relationship between changes in the expression of CYP enzymes in the two experimental models. The obtained results show a fairly similar expression pattern of the main CYP enzymes in the rat livers and lymphocytes, and they indicate that in the liver, lurasidone exerts an inhibitory effect on the activity, protein and mRNA levels of CYP2B1/2 (not CYP2B2 mRNA), CYP2C11 and CYP2E1, while in the case of CYP3A1 and CYP3A2, it causes enzyme induction. At the same time, lurasidone decreases the expression of CYP2B, CYP2C11 (CYP2C11 protein only) and CYP2E1 but increases that of CYP3A2 (not CYP3A1) in lymphocyte cells. In conclusion, chronic treatment with lurasidone simultaneously and in the same way influences the expression and activity of CYP2B, CYP2C11, CYP2E1 and CYP3A2 in the liver and peripheral blood lymphocytes of rats. Thus, the lymphocyte cytochrome P450 profile may be utilized as an indicator of the hepatic cytochrome P450 profile in further clinical studies with lurasidone, and lymphocytes may serve as easily available surrogates for examining the impact of new drugs and chronic in vivo treatments on CYP enzyme expression, as well as to estimate drug-drug interactions and toxicity risk.


Green fluorescent protein-mTalin causes defects in actin organization and cell expansion in Arabidopsis and inhibits actin depolymerizing factor's actin depolymerizing activity in vitro.
Expression of green fluorescent protein (GFP) linked to an actin binding domain is a commonly used method for live cell imaging of the actin cytoskeleton. One of these chimeric proteins is GFP-mTalin (GFP fused to the actin binding domain of mouse talin). Although it has been demonstrated that GFP-mTalin colocalizes with the actin cytoskeleton, its effect on actin dynamics and cell expansion has not been studied in detail. We created Arabidopsis (Arabidopsis thaliana) plants harboring alcohol inducible GFP-mTalin constructs to assess the effect of GFP-mTalin expression in vivo. We focused on the growing root hair as this is a model cell for studying cell expansion and root hair tip growth that requires a highly dynamic and polar actin cytoskeleton. We show that alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization, resulting in either the termination of growth, cell death, and/or changes in cell shape. Fluorescence recovery after photobleaching experiments demonstrate that the interaction of GFP-mTalin and actin filaments is highly dynamic. To assess how GFP-mTalin affects actin dynamics we performed cosedimentation assays of GFP-mTalin with actin on its own or in the presence of the actin modulating protein, actin depolymerizing factor. We show that that GFP-mTalin does not affect actin polymerization but that it does inhibit the actin depolymerizing activity of actin depolymerizing factor. These observations demonstrate that GFP-mTalin can affect cell expansion, actin organization, and the interaction of actin binding proteins with actin.


Modular evolution and the origins of symmetry: reconstruction of a three-fold symmetric globular protein.
The high frequency of internal structural symmetry in common protein folds is presumed to reflect their evolutionary origins from the repetition and fusion of ancient peptide modules, but little is known about the primary sequence and physical determinants of this process. Unexpectedly, a sequence and structural analysis of symmetric subdomain modules within an abundant and ancient globular fold, the β-trefoil, reveals that modular evolution is not simply a relic of the ancient past, but is an ongoing and recurring mechanism for regenerating symmetry, having occurred independently in numerous existing β-trefoil proteins. We performed a computational reconstruction of a β-trefoil subdomain module and repeated it to form a newly three-fold symmetric globular protein, ThreeFoil. In addition to its near perfect structural identity between symmetric modules, ThreeFoil is highly soluble, performs multivalent carbohydrate binding, and has remarkably high thermal stability. These findings have far-reaching implications for understanding the evolution and design of proteins via subdomain modules.


Novel LIPA mutations in Mexican siblings with lysosomal acid lipase deficiency.
Lysosomal acid lipase (LAL) deficiency is an under-recognized lysosomal disease caused by deficient enzymatic activity of LAL. In this report we describe two affected female Mexican siblings with early hepatic complications. At two months of age, the first sibling presented with alternating episodes of diarrhea and constipation, and later with hepatomegaly, elevated transaminases, high levels of total and low-density lipoprotein cholesterol, and low levels of high-density lipoprotein. Portal hypertension and grade 2 esophageal varices were detected at four years of age. The second sibling presented with hepatomegaly, elevated transaminases and mildly elevated low-density lipoprotein and low high-density lipoprotein at six months of age. LAL activity was deficient in both patients. Sequencing of LIPA revealed two previously unreported heterozygous mutations in exon 4: c.253C>A and c.294C>G. These cases highlight the clinical continuum between the so-called Wolman disease and cholesteryl ester storage disease, and underscore that LAL deficiency represents a single disease with a degree of clinical heterogeneity.


Experimental study of fuzzy-rule based management of tropical diseases: case of malaria diagnosis.
The application of the conventional symbolic rules found in knowledge base technology to the management of a disease suffers from its inability to evaluate the degree of severity of a symptom and by extension the degree of the illness. Fuzzy logic technology provides a simple way to arrive at a definite conclusion from vague, ambiguous, imprecise and noisy data (as found in medical data) using linguistic variables that are not necessary precise. In order to achieve this, a study of a knowledge base system for the management of diseases was undertaken. The Root Sum Square of drawing inference was employed to infer the data from the rules developed. This resulted in the establishment of some degrees of influence on the diseases. Using malaria as a case study, a system that uses Visual Basic .Net development environment was developed and the results of the computations are presented in this research.


Identification of Bartonella Species Isolated from Rodents from Yucatan, Mexico, and Isolation of Bartonella vinsonii subsp. yucatanensis subsp. nov.
Bartonella species are highly endemic among wild rodents in many parts of the world. Blood and/or blood clot cultures from 38 rodents, including 27 Yucatan deer mouse (Peromyscus yucatanicus), 7 Gaumer's spiny pocket mouse (Heteromys gaumeri), 2 black rats (Rattus rattus) and 2 big-eared climbing rats (Ototylomys phyllotis) captured near Merida, Yucatan, Mexico, led to the isolation in 3-4 days of small gram-negative bacilli, which were identified as Bartonella spp. based on colony morphology. DNA extraction and PCR testing were also performed from heart samples of 35 of these 38 rodents. Overall, Bartonella spp. were isolated from the blood/blood clots of 22 (58%) rodents. All Bartonella-positive rodents were Yucatán deer mice from San José Pituch. Sequencing of a fragment of the gltA gene showed that all but one rodent isolates were closest to B. vinsonii subsp. vinsonii and one isolate was intermediate between B. vinsonii subsp. berkhoffii and B. vinsonii subsp. arupensis. Further analysis of concatenated housekeeping genes (gltA, ftsZ, rpoB, and 16S rRNA) suggests that this outlier isolate is a new subspecies within the B. vinsonii genogroup, for which we proposed the name B. vinsonii subsp. yucatanensis.


A designed ankyrin repeat protein selected to bind to tubulin caps the microtubule plus end.
Microtubules are cytoskeleton filaments consisting of αβ-tubulin heterodimers. They switch between phases of growth and shrinkage. The underlying mechanism of this property, called dynamic instability, is not fully understood. Here, we identified a designed ankyrin repeat protein (DARPin) that interferes with microtubule assembly in a unique manner. The X-ray structure of its complex with GTP-tubulin shows that it binds to the β-tubulin surface exposed at microtubule (+) ends. The details of the structure provide insight into the role of GTP in microtubule polymerization and the conformational state of tubulin at the very microtubule end. They show in particular that GTP facilitates the tubulin structural switch that accompanies microtubule assembly but does not trigger it in unpolymerized tubulin. Total internal reflection fluorescence microscopy revealed that the DARPin specifically blocks growth at the microtubule (+) end by a selective end-capping mechanism, ultimately favoring microtubule disassembly from that end. DARPins promise to become designable tools for the dissection of microtubule dynamic properties selective for either of their two different ends.