Characterization of dopamine receptors mediating inhibition of adenylate cyclase activity in rat striatum.
In the presence of SCH 23390, a potent blocker of D1 dopamine receptors, dopamine inhibits adenylate cyclase activity of synaptic plasma membranes isolated from rat striatum. Maximal inhibition corresponds to a 20-25% decrease of basal enzyme activity and is reached with 100 microM dopamine. The apparent IC50 of dopamine is 2.5 microM. The inhibitory effect of dopamine is mimicked by various dopamine receptor agonists with the following rank order of potency: (-)-propylnorapomorphine greater than or equal to bromocriptine greater than (+/-)-2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene = (-)-apomorphine greater than dopamine greater than LY 171555 greater than l-noradrenaline greater than l-phenylephrine. Clonidine and l-isoproterenol are inactive at 100 microM. Bromocriptine and LY 171555, two agents which stimulate selectively D2 receptors, inhibit striatal adenylate cyclase activity in the absence of SCH 23390. However, bromocriptine behaves like a partial agonist. A variety of neuroleptic drugs antagonize the dopamine inhibition with a rank order of potency which qualitatively correlates with their relative affinity for D2 receptors. l-Sulpiride (EC50 = 210 nM) and (+)-butaclamol (EC50 = 130 nM) are severalfold more potent than d-sulpiride (EC50 = 5 microM) and (-)-butaclamol (EC50 = 10 microM). The inhibitory effect of dopamine on striatal adenylate cyclase activity is dependent on the presence of GTP, with half-maximal inhibition occurring at 1 microM GTP. In the absence of SCH 23390, dopamine stimulates adenylate cyclase activity, reaching a maximum at 1 microM GTP. At higher concentrations of the nucleotide, the dopamine-stimulated enzyme activity decreases, and this decline is antagonized by the D2 receptor blocker l-sulpiride. Guanyl-5'-yl imidodiphosphate, a stable analogue of GTP, has a biphasic effect on the striatal adenylate cyclase activity, inhibiting at low concentration (from 1 to 100 nM) and stimulating at higher concentrations. Selective activation of D2 receptors by LY 171555 does not increase the extent of enzyme inhibition elicited by guanyl-5'-yl imidodiphosphate. Sodium chloride amplifies the inhibition of striatal adenylate cyclase activity by LY 171555 and reduces the potency of the D2 agonist by a factor of 4. The dopamine-inhibited enzyme activity is lost following intrastriatal injection of kainic acid. The results indicate that in rat striatum dopamine inhibits adenylate cyclase activity by acting on postsynaptic dopamine receptors with pharmacological properties of D2 type.


Segregation of non-p.R132H mutations in IDH1 in distinct molecular subtypes of glioma.
Mutations in the gene encoding the isocitrate dehydrogenase 1 gene (IDH1) occur at a high frequency (up to 80%) in many different subtypes of glioma. In this study, we have screened for IDH1 mutations in a cohort of 496 gliomas. IDH1 mutations were most frequently observed in low grade gliomas with c.395G>A (p.R132H) representing >90% of all IDH1 mutations. Interestingly, non-p.R132H mutations segregate in distinct histological and molecular subtypes of glioma. Histologically, they occur sporadically in classic oligodendrogliomas and at significantly higher frequency in other grade II and III gliomas. Genetically, non-p.R132H mutations occur in tumors with TP53 mutation, are virtually absent in tumors with loss of heterozygosity on 1p and 19q and accumulate in distinct (gene-expression profiling based) intrinsic molecular subtypes. The IDH1 mutation type does not affect patient survival. Our results were validated on an independent sample cohort, indicating that the IDH1 mutation spectrum may aid glioma subtype classification. Functional differences between p.R132H and non-p.R132H mutated IDH1 may explain the segregation in distinct glioma subtypes.


Impact of a mass media campaign to increase public awareness of hypertension.
We evaluated the effect of a nationwide media campaign on hypertension awareness in the population, which was implemented with the aim of spreading key messages related to optimal blood pressure levels and encouraging blood pressure measurements. A nationwide project called "12/8 Awareness Campaign" was implemented between October 2005 and January 2006 using all available mass and outdoor media aiming to improve the knowledge of people on normal blood pressure values and to encourage regular blood pressure measurements. Four survey questions to inquire the level of awareness related to optimal blood pressure levels, hypertension, and hypertension-related disease conditions were directed via face-to-face interviews to two separate samples representing the general Turkish population before (n=1716) and after (n=1725) the campaign, respectively. The answers of the pre- and post-campaign individuals were compared. After the campaign, the percentage of participants who did not know their blood pressure levels decreased from 54.8% to 47.8%, the percentage of those who checked their blood pressure within the past two months increased from 34.3% to 39.6%, and the percentage of those who were aware of the optimal blood pressure levels rose from 51.8% to 58.6% (p<0.001). The campaign contributed significantly to the awareness of hypertension in general population, which is highly encouraging for future efforts for early detection of hypertension and prevention of related morbidity and mortality.


Quantitative analysis of cannabinoids using benchtop NMR instruments.
The quantification of cannabinoids is an essential part of cannabis profiling and testing, whether for medical or recreational use. As regulatory bodies continue to increase testing requirements for these products, it is crucial that alternative and effective analytical methods be developed. Herein, we describe the use of benchtop NMR instruments for the quantification of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) in a variety of cannabis concentrates and compare the values to those obtained using HPLC, the most common approach for the quantification of cannabinoids. Based on the discrepancies observed in test values from different laboratories using only HPLC, the value of orthogonal testing methods has been identified and is increasingly desired.


Energetic and Structural Characterizations of the PET-Water Interface as a Key Step in Understanding Its Depolymerization.
We report molecular simulations of the interaction between poly(ethylene terephthalate) (PET) surfaces and water molecules with a short-term goal to better evaluate the different energy contributions governing the enzymatic degradation of amorphous PET. After checking that the glass transition temperature, density, entanglement mass, and mechanical properties of an amorphous PET are well reproduced by our molecular model, we extend the study to the extraction of a monomer from the bulk surface in different environments, i.e., water, vacuum, dodecane, and ethylene glycol. We complete this energetic characterization by the calculation of the work of adhesion of PET surfaces with water and dodecane molecules and by the determination of the contact angle of water droplets. These calculations are compared with experiments and should help us to better understand the enzymatic degradation of PET from both the thermodynamic and molecular viewpoints.


Identification of novel variants in colorectal cancer families by high-throughput exome sequencing.
Colorectal cancer (CRC) in densely affected families without Lynch Syndrome may be due to mutations in undiscovered genetic loci. Familial linkage analyses have yielded disparate results; the use of exome sequencing in coding regions may identify novel segregating variants. We completed exome sequencing on 40 affected cases from 16 multicase pedigrees to identify novel loci. Variants shared among all sequenced cases within each family were identified and filtered to exclude common variants and single-nucleotide variants (SNV) predicted to be benign. We identified 32 nonsense or splice-site SNVs, 375 missense SNVs, 1,394 synonymous or noncoding SNVs, and 50 indels in the 16 families. Of particular interest are two validated and replicated missense variants in CENPE and KIF23, which are both located within previously reported CRC linkage regions, on chromosomes 1 and 15, respectively. Whole-exome sequencing identified DNA variants in multiple genes. Additional sequencing of these genes in additional samples will further elucidate the role of variants in these regions in CRC susceptibility. Exome sequencing of familial CRC cases can identify novel rare variants that may influence disease risk.


A scalable variational approach to characterize pleiotropic components across thousands of human diseases and complex traits using GWAS summary statistics.
Genome-wide association studies (GWAS) across thousands of traits have revealed the pervasive pleiotropy of trait-associated genetic variants. While methods have been proposed to characterize pleiotropic components across groups of phenotypes, scaling these approaches to ultra large-scale biobanks has been challenging. Here, we propose FactorGo, a scalable variational factor analysis model to identify and characterize pleiotropic components using biobank GWAS summary data. In extensive simulations, we observe that FactorGo outperforms the state-of-the-art (model-free) approach tSVD in capturing latent pleiotropic factors across phenotypes, while maintaining a similar computational cost. We apply FactorGo to estimate 100 latent pleiotropic factors from GWAS summary data of 2,483 phenotypes measured in European-ancestry Pan-UK BioBank individuals (N=420,531). Next, we find that factors from FactorGo are more enriched with relevant tissue-specific annotations than those identified by tSVD (P=2.58E-10), and validate our approach by recapitulating brain-specific enrichment for BMI and the height-related connection between reproductive system and muscular-skeletal growth. Finally, our analyses suggest novel shared etiologies between rheumatoid arthritis and periodontal condition, in addition to alkaline phosphatase as a candidate prognostic biomarker for prostate cancer. Overall, FactorGo improves our biological understanding of shared etiologies across thousands of GWAS.


Creating NADP+ -Specific Formate Dehydrogenases from Komagataella phaffii by Enzymatic Engineering.
Most natural formate dehydrogenases (FDHs) exhibit NAD+ specificity, making it imperative to explore the engineering of FDH cofactor specificity for NADPH regeneration systems. The endogenous FDH of Komagataella phaffii (K. phaffii), termed KphFDH, is a typical NAD+ -specific FDH. However, investigations into engineering the cofactor specificity of KphFDH have yet to be conducted. To develop an NADP+ -specific variant of KphFDH, we selected D195, Y196, and Q197 as mutation sites and generated twenty site-directed variants. Through kinetic characterization, KphFDH/V19 (D195Q/Y196R/Q197H) was identified as the variant with the highest specificity towards NADP+ , with a ratio of catalytic efficiency (kcat /KM )NADP+ /(kcat /KM )NAD+ of 129.226. Studies of enzymatic properties revealed that the optimal temperature and pH for the reduction reaction of NADP+ catalyzed by KphFDH/V19 were 45 °C and 7.5, respectively. The molecular dynamics (MD) simulation was performed to elucidate the mechanism of high catalytic activity of KphFDH/V19 towards NADP+ . Finally, KphFDH/V19 was applied to an in vitro NADPH regeneration system with Meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum (StDAPDH/H227V). This study successfully created a KphFDH variant with high NADP+ specificity and demonstrated its practical applicability in an in vitro NADPH regeneration system.


Differences in glucose values obtained from point-of-care glucose meters and laboratory analysis in critically ill patients.
Blood for glucose analysis is often obtained interchangeably from indwelling catheters and fingersticks. To determine the level of agreement between glucose values obtained by laboratory analysis and with a point-of-care device for blood from 2 different sources: fingerstick and a central venous catheter. A method-comparison design was used. Point-of-care values for blood from fingersticks and catheters were compared with laboratory values for blood from catheters in a convenience sample of 67 critically ill patients. The effects of hematocrit level and finger edema on differences in glucose values between the 2 methods were also evaluated. A t test was used to determine differences in glucose values obtained via the 2 methods. Differences and limits of agreement were also calculated. Laboratory glucose values for blood from a catheter differed significantly from point-of-care values for blood from the catheter (t(1,66) = -9.18; P < .001) and from a fingerstick (t(1,66) = 6.53; P < .001). Glucose values for the 2 methods differed by 20 mg/dL or more for 1 of 6 patients (15%) for catheter samples and for 1 of 5 (21%) for fingerstick samples. Point-of-care glucose values for fingerstick and catheter samples did not differ (P = .98). Hematocrit level significantly explained the difference in glucose values between the 2 methods for both catheter (R(2) = 0.288; P < .001) and fingerstick (R(2) = 0.280; P = .02) samples. Use of a commonly used point-of-care device when precise glucose values are needed may lead to faulty treatment decisions.


High plasma levels of heparin-binding epidermal growth factor are associated with a more stable plaque phenotype and reduced incidence of coronary events.
Rupture of atherosclerotic plaques is the major cause of acute coronary events (CEs). Plaque destabilization is the consequence of an imbalance between inflammatory-driven degradation of fibrous tissue and smooth muscle cell-dependent tissue repair. Proinflammatory factors have been documented extensively as biomarkers of cardiovascular risk but factors that contribute to stabilization of atherosclerotic plaques have received less attention. The present study aimed to investigate whether plasma levels of the smooth muscle cell growth factor epidermal growth factor (EGF), heparin-binding-EGF (HB-EGF), and platelet-derived growth factor correlate with plaque phenotype and incidence of CEs. HB-EGF, EGF and platelet-derived growth factor were measured in plasma from 202 patients undergoing carotid endarterectomy and in 384 incident CE cases and 409 matched controls recruited from the Malmö Diet and Cancer cohort. Significant positive associations were found between the plasma levels of all 3 growth factors and the collagen and elastin contents of the removed plaques. CE cases in the Malmö Diet and Cancer cohort had lower levels of HB-EGF in plasma, whereas no significant differences were found for EGF and platelet-derived growth factor. After adjusting for cardiovascular risk factors in a Cox proportional hazard model, the hazard ratio for the highest HB-EGF tertile was 0.61 (95% confidence interval, 0.47-0.82; P<0.001). The associations between high levels of smooth muscle cell growth factors in plasma and a more fibrous plaque phenotype as well as the association between low levels of HB-EGF and incident CEs point to a potential clinically important role for factors that contribute to plaque stabilization by stimulating smooth muscle cells.


Body mass index and obesity-related metabolic disorders in Taiwanese and US whites and blacks: implications for definitions of overweight and obesity for Asians.
Recommendations based on scanty data have been made to lower the body mass index (BMI; in kg/m(2)) cutoff for obesity in Asians. The goal was to compare relations between BMI and metabolic comorbidity among Asians and US whites and blacks. We compared the prevalence rate, sensitivity, specificity, predictive values, and impact fraction of comorbidities at each BMI level and the BMI-comorbidity relations across ethnic groups by using data from the third National Health and Nutrition Examination Survey and the Nutrition and Health Survey in Taiwan (1993-1996). For most BMI values, the prevalences of hypertension, diabetes, and hyperuricemia were higher for Taiwanese than for US whites. In addition, increments of BMI corresponded to higher odds ratios in Taiwanese than in US whites for hypertriglyceridemia (P = 0.01) and hypertension (P = 0.075). BMI-comorbidity relations were stronger in Taiwanese than in US blacks for all comorbidities studied. BMIs of 22.5, 26, and 27.5 were the cutoffs with the highest sum of positive and negative predictive value for Taiwanese, US white, and US black men, respectively. The same order was observed for women. For BMIs >27, >85% of Taiwanese, 66% of whites, and 55% of blacks had at least one of the studied comorbidities. However, a cutoff close to the median of the studied population was often found by maximizing sensitivity and specificity. Reducing BMI from >25 to <25 in persons in the United States could eliminate 13% of the obesity comorbidity studied. The corresponding cutoff in Taiwan is slightly <24. These data suggest a possible need to set lower BMI cutoffs for Asians, but where to draw the line is a complex issue.


Addressing Profiles of Systemic Inflammation Across the Different Clinical Phenotypes of Acutely Decompensated Cirrhosis.
Background: Patients with acutely decompensated cirrhosis (AD) may or may not develop acute-on-chronic liver failure (ACLF). ACLF is characterized by high-grade systemic inflammation, organ failures (OF) and high short-term mortality. Although patients with AD cirrhosis exhibit distinct clinical phenotypes at baseline, they have low short-term mortality, unless ACLF develops during follow-up. Because little is known about the association of profile of systemic inflammation with clinical phenotypes of patients with AD cirrhosis, we aimed to investigate a battery of markers of systemic inflammation in these patients. Methods: Upon hospital admission baseline plasma levels of 15 markers (cytokines, chemokines, and oxidized albumin) were measured in 40 healthy controls, 39 compensated cirrhosis, 342 AD cirrhosis, and 161 ACLF. According to EASL-CLIF criteria, AD cirrhosis was divided into three distinct clinical phenotypes (AD-1: Creatinine<1.5, no HE, no OF; AD-2: creatinine 1.5-2, and or HE grade I/II, no OF; AD-3: Creatinine<1.5, no HE, non-renal OF). Results: Most markers were slightly abnormal in compensated cirrhosis, but markedly increased in AD. Patients with ACLF exhibited the largest number of abnormal markers, indicating "full-blown" systemic inflammation (all markers). AD-patients exhibited distinct systemic inflammation profiles across three different clinical phenotypes. In each phenotype, activation of systemic inflammation was only partial (30% of the markers). Mortality related to each clinical AD-phenotype was significantly lower than mortality associated with ACLF (p < 0.0001 by gray test). Among AD-patients baseline systemic inflammation (especially IL-8, IL-6, IL-1ra, HNA2 independently associated) was more intense in those who had poor 28-day outcomes (ACLF, death) than those who did not experience these outcomes. Conclusions: Although AD-patients exhibit distinct profiles of systemic inflammation depending on their clinical phenotypes, all these patients have only partial activation of systemic inflammation. However, those with the most extended baseline systemic inflammation had the highest the risk of ACLF development and death.


Dynamics of calcium regulation in Paramecium and possible morphogenetic implication.
This paper is the first report of the use of a fluorescent indicator (Dextran-coupled calcium green-1) for imaging of cytosolic free calcium in ciliate cells. Using this technique in Paramecium, we show that a very transient increase in the mean intracellular calcium concentration accompanied exocytosis. It has long been postulated based on indirect experimental evidence, that a calcium wave which would spread across the cortex at the time of cell division, would be the primary event that triggers morphogenesis in these species. We theoretically show that a unifying interpretation can be given for the possible occurrence of a single wave and that of multiple oscillations of cytosolic calcium: both of which correspond to two different behaviors of the same dynamic system. Experimental conditions allowing the visualization of possible calcium periodicities in the interphase Paramecium cell are much more easily fulfilled than those permitting the observation of a single wave at the time of cell division. Hence, experiments were performed on interphase cells. After microinjection of calcium indicator into a mutant strain which is defective in exocytosis, we observed Ca2+ oscillations with a period close to 2 minutes. Hence, we conclude that Paramecium possesses all the dynamic elements required to generate, at the time of cell division, a morphogenetic calcium wave.


Artificially intelligent chatbots in digital mental health interventions: a review.
Increasing demand for mental health services and the expanding capabilities of artificial intelligence (AI) in recent years has driven the development of digital mental health interventions (DMHIs). To date, AI-based chatbots have been integrated into DMHIs to support diagnostics and screening, symptom management and behavior change, and content delivery. We summarize the current landscape of DMHIs, with a focus on AI-based chatbots. Happify Health's AI chatbot, Anna, serves as a case study for discussion of potential challenges and how these might be addressed, and demonstrates the promise of chatbots as effective, usable, and adoptable within DMHIs. Finally, we discuss ways in which future research can advance the field, addressing topics including perceptions of AI, the impact of individual differences, and implications for privacy and ethics. Our discussion concludes with a speculative viewpoint on the future of AI in DMHIs, including the use of chatbots, the evolution of AI, dynamic mental health systems, hyper-personalization, and human-like intervention delivery.


[Anti-candidal activity of clinical Pseudomonas aeruginosa strains and in vitro inhibition of Candida biofilm formation].
It has been shown recently that signal-mediated interactions between the opportunistic pathogens Pseudomonas aeruginosa and Candida albicans affect virulence features in both organisms. It has been emphasized that the anti-candidal activity of P.aeruginosa is especially due to its pyocyanin pigment. The structure and the properties of monospecies biofilms and their role in infectious disease have been extensively studied. However, there are only few studies on interactions between P.aeruginosa ve Candida spp. İn mixed biofilms. Therefore, in this study, the investigation of anti-candidal activity of P.aeruginosa against Candida spp. And the effect on C.albicans biofilm were aimed. A total of 58 P.aeruginosa strains isolated from several clinical specimens and one of each C.albicans ATCC 90028, C.parapsilosis ATCC 22019, C.glabrata ATCC 90030 and a C.tropicalis clinical isolate were included in the study. The anti-candidal activity of P.aeruginosa strains were investigated on Sabouraud dextrose agar using Kerr's method. The absence of the growth of Candida strains were evaluated as total inhibition. The effect of two P.aeruginosa strains showing total inhibition and one isolate without inhibition effect on C.albicans biofilm was investigated by microplate method. For the evaluation of monospecies or mixed species biofilms of Pseudomonas and C.albicans, the number of colony forming units (cfu/ml) after 90 minutes (adhesion phase), 24 hours and 48 hours (biofilm phase) incubation were determined. Then, cfu numbers of monospecies and mixed species of Pseudomonas and C.albicans were compared. Total inhibition rates of Pseudomonas strains against C.albicans, C.tropicalis, C.glabrata and C.parapsilosis were determined as 63.8%, 75.9%, 65.5% and 72.4%, respectively. A significant reduction was determined in cfu/ml numbers of mixed species of Pseudomonas and C.albicans compared to cfu numbers in monospecies biofilm after 90 minutes, 24 hours and 48 hours (p< 0.05). In addition to the inhibitory effect of P.aeruginosa on Candida growth, in a dual species environment both organisms mutually suppressed each others' biofilm development quantitatively. These findings indicated that molecular basis of signal-mediated interaction between Pseudomonas and Candida spp. Should be clarified for better understanding of the pathogenesis of mixed bacterial-fungal infections.


Finger-Based Numerical Skills Link Fine Motor Skills to Numerical Development in Preschoolers.
Previous studies investigating the association between fine-motor skills (FMS) and mathematical skills have lacked specificity. In this study, we test whether an FMS link to numerical skills is due to the involvement of finger representations in early mathematics. We gave 81 pre-schoolers (mean age of 4 years, 9 months) a set of FMS measures and numerical tasks with and without a specific finger focus. Additionally, we used receptive vocabulary and chronological age as control measures. FMS linked more closely to finger-based than to nonfinger-based numerical skills even after accounting for the control variables. Moreover, the relationship between FMS and numerical skill was entirely mediated by finger-based numerical skills. We concluded that FMS are closely related to early numerical skill development through finger-based numerical counting that aids the acquisition of mathematical mental representations.


MR imaging of common and uncommon large pelvic masses.
Magnetic resonance (MR) imaging is often used in the detection and staging of large pelvic masses. Many large masses in the female pelvis arise from the reproductive organs (eg, uterus, cervix, ovaries, fallopian tubes). In addition, these masses may arise from the gastrointestinal system, urinary system, adjacent soft tissues, peritoneum, or retroperitoneum or from metastases. The majority of large masses in the female pelvis represent such commonly encountered entities as uterine fibroid tumor, dermoid tumor, ovarian cyst, and ovarian cancer. However, uncommon pelvic masses such as mesothelioma, adenocarcinoma, carcinosarcoma, leiomyosarcoma, and desmoid tumor may also be seen. Thus, the differential diagnosis for female pelvic masses is extensive. However, the site of origin, MR imaging characteristics, and clinical history may all help narrow the differential diagnosis. Although with large tumors it may not always be possible to determine the site of origin or distinguish between various tumors at radiology, familiarity with the clinicopathologic and MR imaging features of common and uncommon pelvic masses is important for diagnosis and treatment.


[Phenocopying action of ether on mn d/SM5-strain Drosophila embryos].
Lower sensitivity of the mn d/SM5 strain flies to methyl ether as against the Canton-S flies is established as regarded to induction of bithorax (bx) phenocopies. It is shown that there are no considerable differences in frequencies of bx-phenocopies, abdomen anomalies and ether-induced embryo death between mn d/mn d and mn d/SM5 individuals. It indicates that these parameters are determined by the genotype of the strain on the whole rather than by mn, d, SM5 mutations. Phenocopies of lethal mutations of the pair-rule, Notch, Abnormal abdomen type are established to arise in embryos of both studied strains. In addition disturbances of two other classes are observed among dead embryos of the mn d/SM5 strain. One of them is probably characteristic of SM5/SM5 individuals, the other is phenocopies of mutations disturbing the segment polarity.


In vitro methotrexate polyglutamate synthesis by rat liver folylpolyglutamate synthetase and inhibition by bromosulfophthalein.
We have investigated the properties of the rat liver folylpolyglutamate synthetase using methotrexate (MTX; 4-NH2-10-CH3-PteGlu) as a substrate. Many characteristics of the synthetase (e.g., the apparent Km values for L-glutamate and ATP, and the optimal concentrations of KC1 and 2-mercaptoethanol) are virtually identical whether MTX or tetrahydrofolate is the "folate" substrate. There are, however, several significant differences between the reactions catalyzed with these two substrates. The length of products synthesized from tetrahydrofolate are inversely related to the initial monoglutamate concentration. Low tetrahydrofolate concentrations allow synthesis of longer (n greater than or equal to 3) polyglutamates, up to pentaglutamate length, while high concentrations lead to predominantly diglutamate synthesis. However, 4-NH2-10-CH3-PteGlu2 predominates regardless of the initial MTX concentration, under otherwise identical conditions. Also, tetrahydrofolate can be readily converted to pentaglutamate lengths, the same as predominates in rat liver in vivo. In contrast, MTX forms species containing only up to a total of three glutamates, i.e., 4-NH2-10-CH3-PteGlu3. Finally, the ultimate product of synthesis from tetrahydrofolate, H4PteGlu5, is a fairly good inhibitor of synthetase activity with either MTX or tetrahydrofolate as the substrate. The ultimate product of MTX synthesis, 4-NH2-10-CH3-PteGlu3, however, is a poor inhibitor of activity with either substrate. We have also investigated the inhibitory effect of bromsulfophthalein (BSP) on the rat liver synthetase. Gewirtz et al., showed that this organic anion inhibited the uptake of MTX and 5-CH3-tetrahydrofolate into hepatocytes, and presented indirect evidence that BSP effected polyglutamate biosynthesis. We have demonstrated that BSP is a potent (Ki = 1 microM) inhibitor of the rat liver synthetase. Inhibition is noncompetitive with rspect to L-glutamate, ATP, and MTX. Pre-incubation and time course experiments demonstrated that inhibition is not stoichiometric and is not caused by slow inactivation of the synthetase. Since BSP is transported into mammalian cells, it is the first inhibitor of polyglutamate biosynthesis which has potential for use in in vivo studies.


Crystal structure of an RluF-RNA complex: a base-pair rearrangement is the key to selectivity of RluF for U2604 of the ribosome.
Escherichia coli pseudouridine synthase RluF is dedicated to modifying U2604 in a stem-loop of 23S RNA, while a homologue, RluB, modifies the adjacent base, U2605. Both uridines are in the same RNA stem, separated by approximately 4 A. The 3.0 A X-ray crystal structure of RluF bound to the isolated stem-loop, in which U2604 is substituted by 5-fluorouridine to prevent catalytic turnover, shows RluF distinguishes closely spaced bases in similar environments by a selectivity mechanism based on a frameshift in base pairing. The RNA stem-loop is bound to a conserved binding groove in the catalytic domain. A base from a bulge in the stem, A2602, has folded into the stem, forcing one strand of the RNA stem to translate by one position and thus positioning U2604 to flip into the active site. RluF does not modify U2604 in mutant stem-loops that lack the A2602 bulge and shows dramatically higher activity for a stem-loop with a mutation designed to facilitate A2602 refolding into the stem with concomitant RNA strand translation. Residues whose side chains contact rearranged bases in the bound stem-loop, while conserved among RluFs, are not conserved between RluFs and RluBs, suggesting that RluB does not bind to the rearranged stem loop.


Effect of postextraction algal residue supplementation on the ruminal microbiome of steers consuming low-quality forage.
Cattle consuming low-quality forages (LQF) require protein supplementation to increase forage utilization via ruminal fermentation. Biofuel production from algal biomass results in large quantities of postextraction algal residue (PEAR), which has the potential to elicit LQF utilization responses similar to cottonseed meal (CSM); however, its effect on ruminal bacterial communities is unknown. Five ruminally and duodenally cannulated Angus steers in a 5 × 5 Latin square had ad libitum access to oat straw diets. Treatments were infused ruminally and consisted of an unsupplemented control; PEAR at 50, 100, and 150 mg N/kg BW; and CSM at 100 mg N/kg BW. Ruminal samples were collected 4 h after supplementation on d 14 of each period and separated into solid and liquid fractions. Each sample was extracted for genomic DNA, PCR amplified for the V4 to V6 region of the 16S rRNA, sequenced on the 454 Roche pyrosequencing platform, and analyzed using the QIIME pipeline. Weighted UniFrac analysis and Morisita-Horn index demonstrated different community composition between liquid and solid fractions. Measures of richness including observed operational taxonomic units (OTU) and abundance coverage estimator metric decreased with greater PEAR provision (P ≤ 0.09). There were 42 core microbiome OTU observed in all solid fraction samples while the liquid fraction samples contained 30 core OTU. Bacteroidetes was the predominant phylum followed by Firmicutes in both fractions, which together characterized more than 90% of sequences. Relative abundance of Firmicutes increased with PEAR supplementation in the liquid fraction (linear, P = 0.02). Among Firmicutes, Lachnospiraceae, Ruminococcaceae, and Clostridiaceae families increased in the liquid fraction with greater PEAR supplementation (linear, P ≤ 0.03). Prevotella represented over 25% of sequences in all treatments, and relative abundance decreased in the solid fraction with increasing PEAR provision (linear, P = 0.01). Fibrobacter and Treponema decreased in the liquid fraction with increasing PEAR (linear, P < 0.10). Results suggest PEAR supplementation increased forage utilization by increasing members of Firmicutes within the liquid fraction of the rumen microbiome.


The Duke University Religion Index (DUREL): validation and reliability of the Farsi version.
This study examined the validation and reliability of the Farsi version of the Duke University Religion Index (FDUREL), a brief measure designed to evaluate the primary dimensions of religiosity. The study was conducted in two phases. In the first phase, after translation of the original version of DUREL by using standard forward-backward translation, the FDUREL was administered to 427 medical students at different training levels. Reliability of the FDUREL was assessed by internal consistency and test-retest reliability. Principal components factor analysis was employed to assess the construct validity of the measure. In the second phase, 557 medical students were asked to fill out the FDUREL and Hoge Intrinsic Religiosity Scale to examine concurrent validity. The FDUREL was unidimensional and had good internal consistency and test-retest reliability. Results suggest that the FDUREL is a reliable and valid measure of religiosity in Farsi-speaking populations.


Timing and genetic rapport between growth in skeletal maturity and height around puberty: similarities and differences between girls and boys.
We have analyzed longitudinal twin data by using a multivariate normal model to identify and quantify genetic effects over time on two main aspects of growth, height and skeletal maturity. The largest genetic contribution to the variance in both height and skeletal maturity coincided with the respective ages of peak growth velocity. The highest genetic covariance between these two traits coincided with the age of greatest acceleration of growth in height. These findings imply the existence of regulatory or structural genes that influence growth in both height and skeletal maturity. We also found sex differences in the rapport between velocities for height and skeletal maturity. These are consistent with a predominant role of estrogen in accelerating skeletal maturation in females, and the existence of additional mechanisms in males which may promote growth in height independently of the effects of gonadal sex steroids.


Slow, stochastic transgene repression with properties of a timer.
When gene expression varies unpredictably between genetically identical organisms, this is sometimes ascribed as stochastic. With the prevalence of retroviral vectors, stochastic repression is often observed and can complicate the interpretation of outcomes. But it may also faithfully reflect characteristics of sites in the genome. We created and identified several cell clones in which, within a given cell, retroviral transcription of a transgene was repressed heritably and essentially irreversibly. This repression was relatively slow; total repression in all cells took months. We observed the dynamics of repression and found that they were ergodic, that is, tending with a probability to a final state independent of previous conditions. Different positions of the transgene in the genome demonstrated different dynamics. At a position on mouse chromosome 9, repression abided by near perfect first-order kinetics and was highly reproducible, even under conditions where the number of cell generations per day varied. We propose that such a cell division independent 'off' mechanism could play a role in endogenous gene expression, potentially providing an epigenetically based timer for extended periods.


Exposure of farmers to dust on private farms of various production profiles.
In the light of studies of dustiness in agriculture conducted in Poland and abroad, which deal exclusively with dust risk while performing selected occupations, the results presented in this article are a subsequent attempt to recognize annual exposure to dust at workplaces in agriculture. This recognition concerns the workplace of private farmers--typical of Polish agriculture--on family farms of various profiles of production. The study covered 10 selected farms: four animal breeding farms, four specialised cultivation farms, and two traditional mixed-production farms comprising animal breeding and plant cultivation. The results of the study showed an unequal distribution of the working time load and exposure to dust among the farmers examined during the whole year. The monthly working time limit ranged from 53.6-208.8% of the legal working time. The values of the mean weighted monthly concentrations of total dust ranged within 1.2-33.9 mg x m(-3), and those of respirable dust - 0.3-4.0 mg x m(-3), the highest values being observed in August and September. Mean weighted monthly concentrations describing an average level of farmers' exposure to total dust (7.7-21.9 mg x m(-3)), together with the confidence intervals, remain above the occupational exposure limit (4.0 mg x m(-3)), which is equivalent to hazardous conditions. In the case of respirable dust the results obtained show allowable conditions. Despite this, the dusty working conditions of the farmers in the study should be regarded as hazardous, due to the high level of exposure to total dust observed and potentially high contents of pathogenic components. The highest level of exposure was noted on farms engaged in potato production and marketing, followed by traditional farms carrying out mixed production, while the lowest level of exposure was observed on farms engaged in dairy cattle breeding.


The effect of prematurity on milk composition and its physiological basis.
The composition of milk obtained from mothers giving birth prematurely is of current interest because of the use of this milk as a nutrient source for the premature infant. During the past 5 years research has established that the milk produced by mothers of infants of very low birth weight (e.g., 28-32 wk of gestation) is different from the milk of women giving birth at term. Preterm milk of early lactation is generally found to have higher levels of protein, nonprotein nitrogen, fat, energy, sodium, and chloride, but lower levels of lactose and greater interindividual variation. However, by the end of the first month of lactation the preterm mother's milk is very similar in nutrient composition to term milk, at least based on the incomplete information available at present. Many of the initial differences in composition of preterm compared with term milk may be explained provisionally from animal studies of mammary gland development. Possible explanations include reduced blood flow to the gland and hence milk volume, incomplete differentiation of mammary epithelial cells, and the absence of tight junctions between epithelial cells.


Enhancing esophageal repair with bioactive bilayer mesh containing FGF.
We aimed to prepare a bioactive and biodegradable bilayer mesh formed by fibroblast growth factor (FGF) loaded gelatin film layer, and poly ε-caprolactone (PCL) film layer, and to investigate its treatment efficacy on esophageal anastomosis. It is envisaged that the bioactive mesh in in vivo model would improve tissue healing in rats. The full thickness semicircular defects of 0.5 × 0.5 cm2 were created in anterior walls of abdominal esophagus. The control group had abdominal esophagus isolated with distal esophageal blunt dissection, and sham group had primary anastomosis. In the test groups, the defects were covered with bilayer polymeric meshes containing FGF (5 μg/2 cm2), or not. All rats were sacrificed for histopathology investigation after 7 or 28 days of operation. The groups are coded as FGF(-)-7th day, FGF(+)-7th day, and FGF(+)-28th day, based on their content and operation day. Highest burst pressures were obtained for FGF(+)-7th day, and FGF(+)-28th day groups (p < 0.005) and decreased inflammation grades were observed. Submucosal and muscular collagen deposition scores were markedly increased in these groups compared to sham and FGF(-)-7th day groups having no FGF (p = 0.002, p = 0.001, respectively). It was proved that FGF loaded bioactive bilayer mesh provided effective repair, reinforcement and tissue healing of esophageal defects.


Female dominance in captive gray mouse lemurs (Microcebus murinus).
Female dominance or female feeding priority seem to be characteristic for many lemur species, but are rare traits in other primates and mammals in general. The nocturnal lemur species, however, are underrepresented in the quantitative studies on social dominance. The aim of this study is to investigate the pattern of intersexual dominance relationships in the gray mouse lemur (Microcebus murinus), a species that is generally thought to possess a number of ancestral lemur traits. The context, distribution, and outcome of intersexual conflicts are analyzed in four captive groups of gray mouse lemurs. Intersexual conflicts occurred in the study groups in different behavioral contexts and were mostly spatial interactions (chasing/fleeing, approach/avoidance). The majority of conflicts were decided, and were in all but one case won by females. This is the first evidence suggesting unconditional female dominance in a cheirogaleid primate. The existence of female dominance in most families of the Lemuriformes suggests it is an ancient trait that evolved in their common ancestor.


Postcranial diversity and recent ecomorphic impoverishment of North American gray wolves.
Recent advances in genomics and palaeontology have begun to unravel the complex evolutionary history of the gray wolf, Canis lupus Still, much of their phenotypic variation across time and space remains to be documented. We examined the limb morphology of the fossil and modern North American gray wolves from the late Quaternary (<ca 70 ka) to better understand their postcranial diversity through time. We found that the late-Pleistocene gray wolves were characterized by short-leggedness on both sides of the Cordilleran-Laurentide ice sheets, and that this trait survived well into the Holocene despite the collapse of Pleistocene megafauna and disappearance of the 'Beringian wolf' from Alaska. By contrast, extant populations in the Midwestern USA and northwestern North America are distinguished by their elongate limbs with long distal segments, which appear to have evolved during the Holocene possibly in response to a new level or type of prey depletion. One of the consequences of recent extirpation of the Plains (Canis lupus nubilus) and Mexican wolves (C. l. baileyi) from much of the USA is an unprecedented loss of postcranial diversity through removal of short-legged forms. Conservation of these wolves is thus critical to restoration of the ecophenotypic diversity and evolutionary potential of gray wolves in North America.


D1 versus D2 lymphadenectomy for gastric cancer.
Significant variability exists throughout the world in the extent of lymphadenectomy that is performed for gastric adenocarcinoma. D2 lymphadenectomy is the standard lymphadenectomy performed in high incidence countries such as Japan and South Korea, and less extensive lymphadenectomies are often performed in lower incidence countries such as the Unites States. This article reviews the evidence on the extent of lymphadenectomy that should be performed for gastric adenocarcinoma.


Vitamin D receptor regulates high-level glucose induced retinal ganglion cell damage through STAT3 pathway.
To explore the protective role of Vitamin D Receptor (VDR) in retinal ganglion cells (RGCs) after high-level glucose induction, and to investigate its underlying mechanism. Primary RGCs were isolated from 24-hour-old Sprague Dawley (SD) rats and cultured in 50 mmol/L glucose. The expression of VDR in RGCs induced by 50 mmol/L glucose at different time points was determined by Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, respectively. Subsequently, VDR siRNA was transfected into RGCs. Transfection efficiency was determined by qRT-PCR and Western blot, respectively. The protein expressions levels of VDR, signal transducer and activator of transcription 3 (STAT3) and p-STAT3 in RGCs after VDR knockdown were determined by Western blot. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and cell counting kit-8 (CCK-8) assay were conducted to access the viability of RGCs after high-level glucose induction. Ki-67 staining was performed to detect the proliferation of RGCs. Meanwhile, the apoptosis of RGCs was evaluated by using Annexin-V FITC/PI and TUNEL (terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling) assay, respectively. In addition, caspase-3 activity in RGCs was detected by relative commercial kit. After 4 days of high-level glucose induction, the viability of RGCs was remarkably decreased. VDR was highly expressed in RGCs during high-level glucose culture. The mRNA and protein expression levels of VDR were both significantly downregulated after the transfection of VDR siRNA in RGCs. Meanwhile, VDR knockdown in RGCs significantly increased the viability and proliferative ability of RGCs, whereas significantly decreased apoptotic rate and caspase-3 activity. In addition, the protein level of p-STAT3 in RGCs was remarkably downregulated after VDR knockdown. Inhibition of VDR exerts a protective role in high-level glucose induced RGCs damage by activating the STAT3 pathway.


[Algorithm for radiodiagnosis of blunt neck injury].
The paper analyzes optimal options to use different radiodiagnostic methods for acute blunt injury of the neck. The findings have been used to derive an algorithm for effective radiodiagnostic techniques for blunt neck injury.


Good nutritional control may prevent polyneuropathy after bariatric surgery.
Previously we showed that peripheral neuropathy occurs after bariatric surgery and was associated with malnutrition (mainly sensory polyneuropathy). This study asks whether a multidisciplinary approach to bariatric surgery lowers risk for developing peripheral neuropathy. We performed a retrospective cohort study of all patients with bariatric surgery at the Mayo Clinic between 1985 and 2002. Patients underwent intensive nutritional management before and after surgery. Potential risk factors were analyzed using life-table methods (Cox regression). Univariate analysis showed the following risk factors: increased serum glycosylated hemoglobin and triglycerides, prolonged hospitalization, postoperative gastrointestinal symptoms, and nausea and vomiting. Peripheral neuropathy occurred less frequently (7% vs. 13%, P < 0.01) and specifically the sensory polyneuropathy subtype (1% vs. 7%, P < 0.0001) than in our prior cohort. A systematic, multidisciplinary approach of intensive nutritional management before and after surgery with frequent follow-up greatly decreased development of peripheral neuropathy (especially sensory polyneuropathy) in patients receiving bariatric surgery.


Spike-timing codes enhance the representation of multiple simultaneous sound-localization cues in the inferior colliculus.
To preserve multiple streams of independent information that converge onto a neuron, the information must be re-represented more efficiently in the neural response. Here we analyze the increase in the representational capacity of spike timing over rate codes using sound localization cues as an example. The inferior colliculus receives convergent input from multiple auditory brainstem nuclei, including sound localization information such as interaural level differences (ILDs), interaural timing differences (ITDs), and spectral cues. Virtual space techniques were used to create stimulus sets varying in two sound-localization parameters each. Information about the cues was quantified using a spike distance metric that allows one to separate contributions to the information arising from spike rate and spike timing. Spike timing enhances the representation of spectral and ILD cues at timescales averaging 12 ms. ITD information, however, is carried by a rate code. Comparing responses to frozen and random noise shows that the temporal information is mainly attributable to phase locking to temporal stimulus features, with an additional first-spike latency component. With rate-based codes, there is significant confounding of information about two cues presented simultaneously, meaning that the cues cannot be decoded independently. Spike-timing-based codes reduce this confounded information. Furthermore, the relative representation of the cues often changes as a function of the time resolution of the code, implying that information about multiple cues can be multiplexed onto individual spike trains.


Residual DNA methylation at remission is prognostic in adult Philadelphia chromosome-negative acute lymphocytic leukemia.
Pretreatment aberrant DNA methylation patterns are stable at time of relapse in acute lymphocytic leukemia (ALL). We hypothesized that the detection of residual methylation alterations at the time of morphologic remission may predict for worse prognosis. We developed a real-time bisulfite polymerase chain reaction assay and analyzed the methylation levels of p73, p15, and p57(KIP2) at the time of initial remission in 199 patients with Philadelphia chromosome-negative and MLL(-) ALL. Residual p73 methylation was detected in 18 (9.5%) patients, p15 in 33 (17.4%), and p57(KIP2) in 7 (3.7%); 140 (74%) patients had methylation of 0 genes and 48 (25%) of more than or equal to 1 gene. In 123 (65%) patients, matched pretreatment samples were also studied and compared with remission ones: in 82 of those with initial aberrant methylation of at least one gene, 59 (72%) had no detectable methylation at remission and 23 (28%) had detectable residual methylation. By multivariate analysis, the presence of residual p73 methylation was associated with a significant shorter duration of first complete remission (hazard ratio=2.68, P= .003) and overall survival (hazard ratio=2.69, P= .002). In conclusion, detection of epigenetic alterations allows the identification of patients with ALL with standard risk but with poor prognosis.


Identifying HIV-induced subgraph patterns in brain networks with side information.
Investigating brain connectivity networks for neurological disorder identification has attracted great interest in recent years, most of which focus on the graph representation alone. However, in addition to brain networks derived from the neuroimaging data, hundreds of clinical, immunologic, serologic, and cognitive measures may also be documented for each subject. These measures compose multiple side views encoding a tremendous amount of supplemental information for diagnostic purposes, yet are often ignored. In this paper, we study the problem of subgraph selection from brain networks with side information guidance and propose a novel solution to find an optimal set of subgraph patterns for graph classification by exploring a plurality of side views. We derive a feature evaluation criterion, named gSide, to estimate the usefulness of subgraph patterns based upon side views. Then we develop a branch-and-bound algorithm, called gMSV, to efficiently search for optimal subgraph patterns by integrating the subgraph mining process and the procedure of discriminative feature selection. Empirical studies on graph classification tasks for neurological disorders using brain networks demonstrate that subgraph patterns selected by the multi-side-view-guided subgraph selection approach can effectively boost graph classification performances and are relevant to disease diagnosis.


Clinical and epidemiological profile of cleft lip and palate patients in Peru, 2006 - 2019.
The purpose of this study was to determine the clinical-epidemiological profile of patients with cleft lip and / or palate in Peru from 2006 to 2019. This retrospective and cross-sectional study analyzed 3,923 patients with cleft lip and palate attended by surgical missions of the Operación Sonrisa Perú from January 2006 to December 2019. The clinical profile of the patients treated included: type of cleft (cleft lip CL, cleft palate CP, cleft lip and palate CLP and submucosal SM), surgery performed (cheiloplasty, palatoplasty, cleft rhinoplasty, fistula repair, pharyngeal flap), surgical time according to number of interventions. Likewise, affiliation variables such as sex, age and birthplace were recorded. Descriptive analysis was performed. Associations were determined using the Pearson's Chi-square test and Two-sample test of proportions were used for comparing the percentages during time. A p value <0.005 was considered significant. The most frequent diagnosis was CP (n = 1411, 35.97%). We identified a statistically significant association between the diagnosis of CL, CP and gender (p = 0.045), being more prevalent in males. A higher prevalence of CL was also observed on the left side and in males (n = 183). Cheiloplasty was the most frequent first surgical intervention performed (n = 837, 47.42%) followed by fistula repair as the second intervention (n = 428, 42.29%). Cleft lip and palate are more frequent in males, with CP being the most frequent. CL is more frequent on the left side and the first surgical approach in these patients is lip closure. Key words:Epidemiology, cleft lip and palate, surgical missions.


[Regulation of extracellular phosphohydrolase biosynthesis in bacilli].
Under phosphate-deficient conditions, B. intermedius, B. pumilus, and B. thuringiensis secrete phosphohydrolases, including phosphomono-, phosphodiesterases, and guanyl-specific ribonucleases which cleave RNA molecules to nucleoside-3'-phosphatases. The enzymes are synthesized by phosphate-starved vegetative cells, which is not associated with sporulation. Using B. subtilis strains with mutation in the regulatory protein genes phoP and phoR, it was shown that these proteins regulate expression of B. intermedius, B. pumilus, and B. thuringiensis ribonuclease genes in B. subtilis cells. Genes of heterologous RNAses were activated in recombinant B. subtilis strains simultaneously with its own PHO regulon genes. Presumably a regulatory system homologous to B. subtilis two-component PhoP-PhoR signal transduction system functions in other representatives of the Bacillus genus.


Successful treatment for a patient with hemophagocytic syndrome after a small-for-size graft liver transplantation.
Hemophagocytic syndrome (HPS) is a hematological disorder caused by activated T lymphocytes, which leads to the proliferation of stimulated macrophages that phagocytose and destroy circulating blood elements and their precursors within bone marrow, and lead to the further production of inflammatory cytokines. Living donor liver transplantation (LDLT) between adults has been performed to compensate for the shortage of available organs. There have been some reports concerning HPS after LDLT; however, its prognosis is disappointingly poor. In particular, there is no report of treated HPS developed after LDLT using small-for-size left lobe grafts. We herein report a case of HPS in a 63-year old woman who underwent LDLT using left lobe graft weighing only 330g. The HPS was diagnosed on postoperative day 13, and was successfully treated using a combination of intravenous immunoglobulin, granulocyte colony stimulating factor, conversion of calcineurin inhibitor and steroid pulse. The trigger of HPS may not only be systemic infection, but also hypercytokinemia caused by various factors. HPS is a fatal complication in immunologically compromised patients; however, early and accurate diagnosis could lead to an adequate treatment and improve the outcome.


A new item response theory model for rater centrality using a hierarchical rater model approach.
Rater centrality, in which raters overuse middle scores for rating, is a common rater error which can affect test scores and subsequent decisions. Past studies on rater errors have focused on rater severity and inconsistency, neglecting rater centrality. This study proposes a new model within the hierarchical rater model framework to explicitly specify and directly estimate rater centrality in addition to rater severity and inconsistency. Simulations were conducted using the freeware JAGS to evaluate the parameter recovery of the new model and the consequences of ignoring rater centrality. The results revealed that the model had good parameter recovery with small bias, low root mean square errors, and high test score reliability, especially when a fully crossed linking design was used. Ignoring centrality yielded poor item difficulty estimates, person ability estimates, rater errors estimates, and underestimated reliability. We also showcase how the new model can be used, using an empirical example involving English essays in the Advanced Placement exam.


The obese brain athlete: self-regulation of the anterior insula in adiposity.
The anterior insular cortex (AIC) is involved in emotional processes and gustatory functions which can be examined by imaging techniques. Such imaging studies showed increased activation in the insula in response to food stimuli as well as a differential activation in lean and obese people. Additionally, studies investigating lean subjects established the voluntary regulation of the insula by a real-time functional magnetic resonance imaging-brain computer interface (rtfMRI-BCI) approach. In this exploratory study, 11 lean and 10 obese healthy, male participants were investigated in a rtfMRI-BCI protocol. During the training sessions, all obese participants were able to regulate the activity of the AIC voluntarily, while four lean participants were not able to regulate at all. In successful regulators, functional connectivity during regulation vs. relaxation between the AIC and all other regions of the brain was determined by a seed voxel approach. Lean in comparison to obese regulators showed stronger connectivity in cingular and temporal cortices during regulation. We conclude, that obese people possess an improved capacity to self-regulate the anterior insula, a brain system tightly related to bodily awareness and gustatory functions.


Calculations on tunneling in the reactions of noradamantyl carbenes.
Noradamantylchlorocarbene has been found experimentally to undergo ring expansion to 2-chloroadamantene at cryogenic temperatures. The rate constant, calculated with inclusion of small-curvature tunneling, is within a factor of 2 of the rate constant measured at 9 K in a nitrogen matrix. Our calculations predict that noradamantylfluorocarbene will not be found to rearrange under these conditions. The rate constant for carbon tunneling in the ring expansion of noradamantylmethylcarbene (1d) to 2-methyladamantene at T </~ 10 K is calculated to be lower by more than 8 orders of magnitude than the rate constant for formation of 3-vinylnoradamantane from 1d by hydrogen migration.


Sensorineural hearing loss and Kawasaki disease: a prospective study.
Kawasaki disease (KD) is an acute, self-limited vasculitis of infants and children that is now the most common cause of acquired heart disease in the pediatric age group in the United States and Japan. Reports have documented the association of acute KD with sensorineural hearing loss. To assess the prevalence of hearing loss following acute KD in a geographically and ethnically diverse population, a prospective, multicenter study of hearing loss in patients with KD was conducted. Patients with acute KD were enrolled in 7 clinical centers and underwent a primary audiologic evaluation within 30 days of the onset of fever. Patients were subsequently reevaluated after resolution of the acute phase of the disease. A questionnaire assessing risk factors for hearing loss was also administered. A total of 62 patients were evaluated during the 29-month study period. At the first audiologic evaluation, 19 patients (30.6%) had sensorineural hearing loss, 6 patients (9.7%) had conductive hearing loss, 17 patients (27.4%) had normal hearing, and 20 patients (32.3%) had inconclusive studies. Overall, 2 of 36 patients (5.5%) had sensorineural hearing loss documented on their second audiologic evaluation. No risk factors for hearing loss were identified by the questionnaire. Transient sensorineural hearing loss (20 to 35 dB) is a frequent complication of acute KD and may be related to salicylate toxicity in some patients. Persistent sensorineural hearing loss is uncommon. Parents and primary care providers should be made aware of the potential for persistent sensorineural hearing loss following resolution of KD, but routine audiologic screening of this patient population does not appear to be warranted.


Photonic Dirac cavities with spatially varying mass term.
In recent years, photonics has proven itself as an excellent platform for emulation of relativistic phenomena. Here, we show an example of relativistic-like trapping in photonic system that realizes Dirac-like dispersion with spatially inhomogeneous mass term. The modes trapped by such cavities, their energy levels, and corresponding orbitals are then characterized through optical imaging in real and momentum space. The fabricated cavities host a hierarchy of photonic modes with distinct radiation profiles directly analogous to various atomic orbitals endowed with unique characteristics, such as pseudo-particle-hall symmetry and spin degeneracy, and they carry topological charge which gives rise to radiative profiles with angular momentum. We demonstrate that these modes can be directionally excited by pseudo-spin-polarized boundary states. In addition to the fundamental interest in the structure of these pseudo-relativistic orbitals, the proposed system offers a route for designing new types of nanophotonic devices, spin-full resonators and topological light sources compatible with integrated photonics platforms.


How to implement patient experience surveys and use their findings for service improvement: a qualitative expert consultation study in Australian general practice.
To identify barriers (patient, provider, practice and system levels) to consider when implementing patient experience surveys in Australian general practice and enablers of their systematic use to inform service improvement in clinical practice as well as the broader health system. An expert consultation and qualitative content analysis of cross-sectional, open-text survey data. Data were collected from key international and Australian experts in the areas of measurement and quality improvement in general practice. Responses from 20 participants from six countries were included in the study. Participants discussed the importance of ensuring value and relevance of surveys to stakeholders. Lack of resources, IT infrastructure, capacity building and sustained funding were identified as barriers to implementing surveys. Participants discussed the importance of clearly defining and communicating the purpose of surveys and agreed on the value of using patient experience to inform reflective, team-based learning at the practice level. Opinions differed on the use of patient experience data at the system level, with some questioning its utility or fairness for external performance reporting. Others recommended the aggregation and reporting of these data under certain conditions, including for the purpose of triangulation with other quality and outcome data. The study identified an evidence gap in the assessment and interpretation of patient experience data at the practice and system levels, including the analysis and contextualisation of survey findings at the system level. Patient experience surveys have potential for guiding practice level quality improvement, but many barriers to their implementation remain. There is need for greater research and policy efforts to understand how this information can be used at the system level for improving Australian general practice.


Linkage of Infection to Adverse Systemic Complications: Periodontal Disease, Toll-Like Receptors, and Other Pattern Recognition Systems.
Toll-like receptors (TLRs) are a group of pattern recognition receptors (PRRs) that provide innate immune sensing of conserved pathogen-associated molecular patterns (PAMPs) to engage early immune recognition of bacteria, viruses, and protozoa. Furthermore, TLRs provide a conduit for initiation of non-infectious inflammation following the sensing of danger-associated molecular patterns (DAMPs) generated as a consequence of cellular injury. Due to their essential role as DAMP and PAMP sensors, TLR signaling also contributes importantly to several systemic diseases including cardiovascular disease, diabetes, and others. The overlapping participation of TLRs in the control of infection, and pathogenesis of systemic diseases, has served as a starting point for research delving into the poorly defined area of infection leading to increased risk of various systemic diseases. Although conflicting studies exist, cardiovascular disease, diabetes, cancer, rheumatoid arthritis, and obesity/metabolic dysfunction have been associated with differing degrees of strength to infectious diseases. Here we will discuss elements of these connections focusing on the contributions of TLR signaling as a consequence of bacterial exposure in the context of the oral infections leading to periodontal disease, and associations with metabolic diseases including atherosclerosis and type 2 diabetes.


Genistein modulation of streptozotocin diabetes in male B6C3F1 mice can be induced by diet.
Diet and phytoestrogens affect the development and progression of diabetes. The objective of the present study was to determine if oral exposure to phytoestrogen genistein (GE) by gavage changed blood glucose levels (BGL) through immunomodulation in streptozotocin (STZ)-induced diabetic male B6C3F1 mice fed with three different diets. These three diets were: NTP-2000 diet (NTP), soy- and alfalfa-free 5K96 diet (SOF) and high fat diet (HFD) with 60% of kcal from fat, primarily rendered fat of swine. The dosing regimen for STZ consisted of three 100mg/kg doses (i.p.): the first dose was administered at approximately 2weeks following the initiation of daily GE (20mg/kg) gavage, and the second dose was on day 19 following the first dose, and the third dose was on day 57 following the first dose. In mice on the NTP diet, GE treatment decreased BGL with statistical significances observed on days 33 and 82 following the first STZ injection. In mice fed the HFD diet, GE treatment produced a significant decrease and a significant increase in BGL on days 15 and 89 following the first STZ injection, respectively. In mice fed the SOF diet, GE treatment had no significant effects on BGL. Although GE treatment affected phenotypic distributions of both splenocytes (T cells, B cells, natural killer cells and neutrophils) and thymocytes (CD4/CD8 and CD44/CD25), and their mitochondrial transmembrane potential and generation of reactive oxygen species, indicators of cell death (possibly apoptosis), GE modulation of neutrophils was more consistent with its diabetogenic or anti-diabetic potentials. The differential effects of GE on BGL in male B6C3F1 mice fed with three different diets with varied phytoestrogen contents suggest that the estrogenic properties of this compound may contribute to its modulation of diabetes.


The effect of HMO penetration on physician retirement.
To examine the effect of HMO penetration on physician retirement. We linked together historical data from the Physician Masterfile of the American Medical Association for successive years to track changes in physicians' activity status between 1980 and 1997. We used a multivariate discrete-time survival model to examine how the probability of physician retirement was affected by the level of HMO penetration in the physician's market area, controlling for other physician and market characteristics. The study population included all active allopathic patient-care physicians in the United States who reached age 55 between the years of 1980 and 1996. The main outcome measure was physician retirements as reported on the Physician Masterfile. HMO penetration had a statistically significant positive effect on the retirement probabilities of generalists and medical/surgical specialists, but it s effect on hospital-based specialists and psychiatrists was not significant . For generalists regression-adjusted retirement probabilities were roughly 13 percent greater in high-penetration markets (HMO penetration of 45 percent ) than in low-penetration markets (HMO penetration of 5 percent ). For medical/surgical specialist s regression-adjusted retirement probabilities were roughly 17 percent greater in high-penetration markets than in low-penetration markets. Our findings suggest that many older physicians have found it preferable to retire rather than adapt their practices to an environment with a high degree of managed care penetration . Because the number of physicians entering the older age categories will increase rapidly over the next 20 years, the growth of managed care and other influences on physician retirement will play an increasingly important role in determining the size of the physician workforce.


Investigation of Avian Influenza H5N6 Virus-like Particles as a Broad-Spectrum Vaccine Candidate against H5Nx Viruses.
Highly pathogenic avian influenza (HPAI) clade 220.127.116.11 viruses have been reported to be the source of infections in several outbreaks in the past decades. In a previous study, we screened out a broad-spectrum virus strain, H5N6-Sichuan subtype, by using a lentiviral pseudovirus system. In this project, we aimed to investigate the potential of H5N6 virus-like particles (VLPs) serving as a broad-spectrum vaccine candidate against H5Nx viruses. We cloned the full-length M1 gene and H5, N6 genes derived from the H5N6-Sichuan into pFASTBac vector and generated the VLPs using the baculovirus-insect cell system. H5N6 VLPs were purified by sucrose gradient centrifugation, and the presence of H5, N6 and M1 proteins was verified by Western blot and SDS-PAGE. The hemagglutination titer of H5N6 VLPs after purification reached 5120 and the particle structure remained as viewed by electron microscopy. The H5N6 VLPs and 293T mammalian cell-expressed H5+N6 proteins were sent for mice immunization. Antisera against the H5+N6 protein showed 80 to 320 neutralizing antibody titers to various H5Nx pseudoviruses. In contrast, H5N6 VLPs not only elicited higher neutralizing antibody titers, ranging from 640 to 1280, but also induced higher IL-2, IL-4, IL-5, IFN-γ and TNF production, thus indicating that H5N6 VLPs may be a potential vaccine candidate for broad-spectrum H5Nx avian influenza vaccines.


[Structural, functional and molecular characteristics of the central nervous system of young fish raised in ecologically different conditions].
On example of wild and hatchery (cultivated in ponds and pools) young Russian sturgeons and white salmons, it is shown that fishes cultivated in more complicated ecological conditions (in natural reservoir in comparison to artificial and in ponds in comparison to pools) have greater sizes of CNS structures connected with the development of the analyzer leading in the life of this species (the midbrain--in white salmon, the forebrain--in sturgeon) and are characterized by a higher content of DNA in the brain and of RNA at the unit of DNA. This testified to more intensive processes of replication and transcription in CNS cells, more expressed manifestation of motor activity and higher parameters of elaboration and retention of a new habit. All this points to the necessity of enrichment of the medium in the hatchery reservoirs and the expediency of using of genetic-neurobiological criteria of hatching the young.


Reducing the risk for distal forearm fracture: preserve bone mass, slow down, and don't fall!
A case-control study of 1,150 female and male distal forearm cases and 2,331 controls of age 45 years and older was undertaken from 1996-2001 in five Northern California Kaiser Permanente Medical Centers. Most information on possible risk factors was obtained by an interviewer-administered questionnaire, supplemented by a few tests of lower extremity neurological function. Previous fractures since 45 years of age, a rough marker of osteoporosis, were associated with an increased risk (adjusted odds ratio [OR] [95% confidence interval] = 1.48 [1.20-1.84 ] per previous fracture). Several factors thought to protect against low bone mass were associated with a reduced risk, including current use of menopausal hormone therapy (adjusted OR = 0.60 [0.49-0.74]), ever used thiazide diuretics or water pills for at least 1 year (adjusted OR = 0.79 [0.64-0.97]), high body mass index (weight in kg/height in m2) (adjusted OR = 0.96 [0.89-1.04] per 5 unit increase), and high dietary calcium intake (adjusted OR = 0.88 [0.75-1.03] per 500 mg/day). Falls in the past year and conditions associated with falling, such as epilepsy and/or use of seizure medication (adjusted OR = 2.07 [1.35-3.17]) and a history of practitioner-diagnosed depression (adjusted OR = 1.40 [1.13-1.73]), were associated with increased risks. Having difficulty performing physical functions and all lower-extremity problems measured in this study were associated with reduced risks. The results from this and other studies indicate that distal forearm fractures tend to occur in people with low bone mass who are otherwise in relatively good health and are physically active, but who are somewhat prone to falling (particularly on an outstretched hand), and whose movements are not slowed by lower extremity problems and other debilities. Thus, measures to decrease fall frequency and to slow down the pace of relatively healthy people with low bone mass should lead to a lower frequency of distal forearm fracture.


Pseudomonas aeruginosa antibodies in blood spots from patients with cystic fibrosis.
The formation of antibodies to Pseudomonas aeruginosa may be the earliest indicator of pulmonary infection in patients with cystic fibrosis. To enable easy sampling in babies and young children an enzyme linked immunosorbent assay (ELISA) based on a blood spot sample taken on to blotting paper was developed. A sample of approximately 20 microliters of blood was required. A high correlation and level of absolute agreement was shown between paired finger prick and venepuncture blood spots, and between blood spot, serum spot, and serum samples. Healthy controls and non-infected patients with cystic fibrosis had low titres of antibody compared with patients with intermittent and chronic infection. The latter groups had significantly greater antibody titres than normal controls. This assay permits serial measurement of antibodies to P aeruginosa in patients of all ages with cystic fibrosis and may provide a means of assessing the value of such measurements in the detection and management of early infection.


Human rights, health and the state in Bangladesh.
This paper broadly discusses the role of the State of Bangladesh in the context of the health system and human rights. The interrelation between human rights, health and development are well documented. The recognition of health as a fundamental right by WHO and subsequent approval of health as an instrument of welfare by the Universal Declaration of Human Rights (UDHR) and the International Covenant on Social, Economic and Cultural Rights (ICSECR) further enhances the idea. Moreover, human rights are also recognized as an expedient of human development. The state is entrusted to realize the rights enunciated in the ICSECR. In exploring the relationship of the human rights and health situation in Bangladesh, it is argued, in this paper, that the constitution and major policy documents of the Bangladesh government have recognized the health rights and development. Bangladesh has ratified most of the international treaties and covenants including ICCPR, ICESCR; and a signatory of international declarations including Alma-Ata, ICPD, Beijing declarations, and Millennium Development Goals. However the implementation of government policies and plans in the development of health institutions, human resources, accessibility and availability, resource distribution, rural-urban disparity, the male-female gap has put the health system in a dismal state. Neither the right to health nor the right to development has been established in the development of health system or in providing health care. The development and service pattern of the health system have negative correlation with human rights and contributed to the underdevelopment of Bangladesh. The government should take comprehensive approach in prioritizing the health rights of the citizens and progressive realization of these rights.


[The power of the notion of resistance in the mental health field: a case study on the life of rural communities whitin the Colombian armed conflict].
The objective of this study is to identify individual, family and/or community responses of resistance within protracted armed conflict. We conducted a case study with members of rural communities in the municipality of San Francisco, in the eastern area of Antioquia, Colombia, combining biographical and ethnographical approaches. The primary results show that, along with the suffering generated by the experience of armed conflict, rural community members also display a repertoire of multiple and diverse resistance strategies. Resistance is for them an active response and a way to re-weave the fabric torn by the experience. As a type of affirmative opposition, resistance is a powerful category for the entire mental health field, in that it highlights the creativity and capacity for transformation of individuals. In this way, the category allows for overcoming the limits of the conventional biomedical view that tends to pathologize individual and social responses in scenarios of severe distress.


Non-visual exploration of novel objects increases the levels of plasticity factors in the rat primary visual cortex.
Historically, the primary sensory areas of the cerebral cortex have been exclusively associated with the processing of a single sensory modality. Yet the presence of tactile responses in the primary visual (V1) cortex has challenged this view, leading to the notion that primary sensory areas engage in cross-modal processing, and that the associated circuitry is modifiable by such activity. To explore this notion, here we assessed whether the exploration of novel objects in the dark induces the activation of plasticity markers in the V1 cortex of rats. Adult rats were allowed to freely explore for 20 min a completely dark box with four novel objects of different shapes and textures. Animals were euthanized either 1 (n = 5) or 3 h (n = 5) after exploration. A control group (n = 5) was placed for 20 min in the same environment, but without the objects. Frontal sections of the brains were submitted to immunohistochemistry to measure protein levels of egr-1 and c-fos, and phosphorylated calcium-dependent kinase (pCaKMII) in V1 cortex. The amount of neurons labeled with monoclonal antibodies against c-fos, egr-1 or pCaKMII increased significantly in V1 cortex after one hour of exploration in the dark. Three hours after exploration, the number of labeled neurons decreased to basal levels. Our results suggest that non-visual exploration induces the activation of immediate-early genes in V1 cortex, which is suggestive of cross-modal processing in this area. Besides, the increase in the number of neurons labeled with pCaKMII may signal a condition promoting synaptic plasticity.


A new pleiotropic effect of statins in elderly: modulation of telomerase activity.
Recent evidence suggests a link between statins and telomere biology. Whether statin treatment may modulate telomerase activity and affect telomere erosion rate is unknown. We aimed at investigating the potential impact of statin therapy on peripheral blood mononuclear cells telomerase activity, its implication on LTL variability, and its association with telomere shortening rates along with aging. The cross-sectional study was conducted in 230 subjects (age range: 30-86 y) stratified according to statins treatment. LTL was measured by quantitative polymerase chain reaction and telomerase activity by a PCR-ELISA protocol. Subjects on statin treatment showed higher telomerase activity (P<0.0001) and longer LTL (P=0.028) levels compared to the nonstatin group. Statin therapy was associated with higher telomerase activity independently of multiple covariates, including age, gender, smoking habits, lipid, systemic inflammation, glucose, and blood pressure levels (P=0.019). Indeed, subjects on statin treatment showed significant lower telomere erosion along with aging. Every 1 y increment in age, LTL decreases by 0.058 Kb in no statin and 0.033 Kb in statin groups, respectively, as well as the major difference in telomere attrition between groups was found after the age of 65 yr (P<0.0001). In summary, statins, modulating telomerase activity, affect telomere erosion along with aging.


An epidural initial dose is unnecessary in combined spinal epidural anesthesia for Caesarean section.
Combined spinal epidural anesthesia is widely used for Caesarean section. Bolus administration of an epidural initial dose introduces the risk of drug flux from the epidural space to the subarachnoid space, and the volume effect of the initial dose may cause epidural top-up and extension of subarachnoid blockade. These problems may be avoided if the initial dose is not administered. This study investigated whether epidural continuous infusion without an initial dose (continuous group) can decrease postoperative pain as well as an epidural continuous infusion with an initial dose (initial dose group). Sixty-one patients undergoing elective Caesarean section were randomly assigned to the initial dose group or the continuous group. Twenty patients undergoing emergency Caesarean section with spinal anesthesia (spinal group) were also investigated to confirm that epidural block is effective for postoperative pain. Data in this study were obtained retrospectively from each patient's records. Between the initial dose group and the continuous group, there was no significant difference in the number of times flurbiprofen or pentazocine were used for postoperative pain relief. However, the number of times that pentazocine was used was significantly higher in the spinal group than in other groups. This finding suggests that an epidural initial dose is unnecessary for postoperative pain relief in combined spinal epidural anesthesia for Caesarean section.


Vascular calcification and left ventricular hypertrophy in hemodialysis patients: interrelationship and clinical impacts.
Background: We examined the relationship and combined effect of vascular calcification (VC) and left ventricular hypertrophy (LVH) on deaths and cardiovascular events (CVEs) in hemodialysis (HD) patients. Methods: Maintenance HD patients (n=341) were included. Echocardiography data and plain chest radiographs were used to assess LVH and aortic arch VC. Results: VC was found in 100 patients (29.3%). LVH was more prevalent in patients with VC compared with those without VC (70% vs. 50.2%, P=0.001). VC was independently associated with a 2.42-fold increased risk of LVH (95% CI, 1.26-4.65). In multivariate analysis, compared with patients with neither VC nor LVH, the coexistence of VC and LVH was independently associated with CVE (HR, 2.01; 95% CI, 1.09-3.72), whereas VC or LVH alone was not. Patients with both VC and LVH had the highest risk for a composite event of deaths or CVE (HR, 1.88; 95% CI, 1.15-3.06). Significant synergistic interaction was observed between VC and LVH (P for interaction=0.039). Conclusions: VC was independently associated with LVH. The coexistence of VC and LVH was associated with higher risk of deaths and CVEs than either factor alone. VC and LVH showed a synergistic interaction for the risk of deaths and CVEs.


Tiletamine is a potent inhibitor of N-methyl-aspartate-induced depolarizations in rat hippocampus and striatum.
N-methyl-D,L-aspartate (NMA) antagonists are of potential value in the treatment of epilepsy and ischemia, but commonly utilized compounds are of low potency and poorly penetrate the brain. Tiletamine hydrochloride is a lipophilic and potent veterinary anesthetic. This study shows tiletamine to be similar to ketamine and to phencyclidine, agents known to interact with the NMA receptor. Effects of tiletamine on synaptic transmission and on direct excitatory responses to exogenous amino acids were examined in rat hippocampal and striatal slices. In striatal slices, tiletamine inhibited the NMA-mediated, but not the spontaneous, release of [3H]acetylcholine, with an IC50 of 70 nM. In hippocampal CA1 cells, 3 microM tiletamine in the perfusate reversibly blocked the intracellularly recorded responses to ionophoretically applied NMA, but not to glutamate, quisqualate and kainate. Tiletamine, 3 to 100 microM, had no effect on the orthodromically elicited excitatory postsynaptic potential, action potential amplitude or duration, resting membrane potential, or input resistance. In Mg++-free perfusate, the excitatory postsynaptic potential was greatly augmented to give a paroxysmal depolarization shift and was reversibly blocked by 10 microM tiletamine. Our results show that tiletamine is a potent and reversible antagonist of NMA-mediated responses without itself having major effects in low concentrations on normal membrane and synaptic pyramidal cell properties.


Identifying correlates of Guinea worm (Dracunculus medinensis) infection in domestic dog populations.
Few human infectious diseases have been driven as close to eradication as dracunculiasis, caused by the Guinea worm parasite (Dracunculus medinensis). The number of human cases of Guinea worm decreased from an estimated 3.5 million in 1986 to mere hundreds by the 2010s. In Chad, domestic dogs were diagnosed with Guinea worm for the first time in 2012, and the numbers of infected dogs have increased annually. The presence of the parasite in a non-human host now challenges efforts to eradicate D. medinensis, making it critical to understand the factors that correlate with infection in dogs. In this study, we evaluated anthropogenic and environmental factors most predictive of detection of D. medinensis infection in domestic dog populations in Chad. Using boosted regression tree models to identify covariates of importance for predicting D. medinensis infection at the village and spatial hotspot levels, while controlling for surveillance intensity, we found that the presence of infection in a village was predicted by a combination of demographic (e.g. fishing village identity, dog population size), geographic (e.g. local variation in elevation), and climatic (e.g. precipitation and temperature) factors, which differed between northern and southern villages. In contrast, the presence of a village in a spatial infection hotspot, was primarily predicted by geography and climate. Our findings suggest that factors intrinsic to individual villages are highly predictive of the detection of Guinea worm parasite presence, whereas village membership in a spatial infection hotspot is largely determined by location and climate. This study provides new insight into the landscape-scale epidemiology of a debilitating parasite and can be used to more effectively target ongoing research and possibly eradication and control efforts.


Scepticism about the virtue ethics approach to nursing ethics.
Nursing ethics centres on how nurses ought to respond to the moral situations that arise in their professional contexts. Nursing ethicists invoke normative approaches from moral philosophy. Specifically, it is increasingly common for nursing ethicists to apply virtue ethics to moral problems encountered by nurses. The point of this article is to argue for scepticism about this approach. First, the research question is motivated by showing that requirements on nurses such as to be kind, do not suffice to establish virtue ethics in nursing because normative rivals (such as utilitarians) can say as much; and the teleology distinctive of virtue ethics does not transpose to a professional context, such as nursing. Next, scepticism is argued for by responding to various attempts to secure a role for virtue ethics in nursing. The upshot is that virtue ethics is best left where it belongs - in personal moral life, not professional ethics - and nursing ethics is best done by taking other approaches.


Quantitative effects of P elements on hybrid dysgenesis in Drosophila melanogaster.
Genetic analyses involving chromosomes from seven inbred lines derived from a single M' strain were used to study the quantitative relationships between the incidence and severity of P-M hybrid dysgenesis and the number of genomic P elements. In four separate analyses, the mutability of snw, a P element-insertion mutation of the X-linked singed locus, was found to be inversely related to the number of autosomal P elements. Since snw mutability is caused by the action of the P transposase, this finding supports the hypothesis that genomic P elements titrate the transposase present within a cell. Other analyses demonstrated that autosomal transmission ratios were distorted by P element action. In these analyses, the amount of distortion against an autosome increased more or less linearly with the number of P elements carried by the autosome. Additional analyses showed that the magnitude of this distortion was reduced when a second P element-containing autosome was present in the genome. This reduction could adequately be explained by transposase titration; there was no evidence that it was due to repressor molecules binding to P elements and inhibiting their movement. The influence of genomic P elements on the incidence of gonadal dysgenesis was also investigated. Although no simple relationship between the number of P elements and the incidence of the trait could be discerned, it was clear that even a small number of elements could increase the incidence markedly. The failure to find a quantitative relationship between P element number and the incidence of gonadal dysgenesis probably reflects the complex etiology of this trait.


Design and performance evaluation of a coplanar multimodality scanner for rodent imaging.
This work reports on the development and performance evaluation of the VrPET/CT, a new multimodality scanner with coplanar geometry for in vivo rodent imaging. The scanner design is based on a partial-ring PET system and a small-animal CT assembled on a rotatory gantry without axial displacement between the geometric centers of both fields of view (FOV). We report on the PET system performance based on the NEMA NU-4 protocol; the performance characteristics of the CT component are not included herein. The accuracy of inter-modality alignment and the imaging capability of the whole system are also evaluated on phantom and animal studies. Tangential spatial resolution of PET images ranged between 1.56 mm at the center of the FOV and 2.46 at a radial offset of 3.5 cm. The radial resolution varies from 1.48 mm to 1.88 mm, and the axial resolution from 2.34 mm to 3.38 mm for the same positions. The energy resolution was 16.5% on average for the entire system. The absolute coincidence sensitivity is 2.2% for a 100-700 keV energy window with a 3.8 ns coincident window. The scatter fraction values for the same settings were 11.45% for a mouse-sized phantom and 23.26% for a rat-sized phantom. The peak noise equivalent count rates were also evaluated for those phantoms obtaining 70.8 kcps at 0.66 MBq/cc and 31.5 kcps at 0.11 MBq/cc, respectively. The accuracy of inter-modality alignment is below half the PET resolution, and the image quality of biological specimens agrees with measured performance parameters. The assessment presented in this study shows that the VrPET/CT system is a good performance small-animal imager, while the cost derived from a partial ring detection system is substantially reduced as compared with a full-ring PET tomograph.


ActRIIB:ALK4-Fc alleviates muscle dysfunction and comorbidities in murine models of neuromuscular disorders.
Patients with neuromuscular disorders suffer from a lack of treatment options for skeletal muscle weakness and disease comorbidities. Here, we introduce as a potential therapeutic agent a heterodimeric ligand-trapping fusion protein, ActRIIB:ALK4-Fc, which comprises extracellular domains of activin-like kinase 4 (ALK4) and activin receptor type IIB (ActRIIB), a naturally occurring pair of type I and II receptors belonging to the TGF-β superfamily. By surface plasmon resonance (SPR), ActRIIB:ALK4-Fc exhibited a ligand binding profile distinctly different from that of its homodimeric variant ActRIIB-Fc, sequestering ActRIIB ligands known to inhibit muscle growth but not trapping the vascular regulatory ligand bone morphogenetic protein 9 (BMP9). ActRIIB:ALK4-Fc and ActRIIB-Fc administered to mice exerted differential effects - concordant with SPR results - on vessel outgrowth in a retinal explant assay. ActRIIB:ALK4-Fc induced a systemic increase in muscle mass and function in wild-type mice and in murine models of Duchenne muscular dystrophy (DMD), amyotrophic lateral sclerosis (ALS), and disuse atrophy. Importantly, ActRIIB:ALK4-Fc improved neuromuscular junction abnormalities in murine models of DMD and presymptomatic ALS and alleviated acute muscle fibrosis in a DMD model. Furthermore, in combination therapy ActRIIB:ALK4-Fc increased the efficacy of antisense oligonucleotide M12-PMO on dystrophin expression and skeletal muscle endurance in an aged DMD model. ActRIIB:ALK4-Fc shows promise as a therapeutic agent, alone or in combination with dystrophin rescue therapy, to alleviate muscle weakness and comorbidities of neuromuscular disorders.


Influence of glycosaminoglycan identity on vocal fold fibroblast behavior.
Poly(ethylene glycol) (PEG) hydrogels have recently begun to be studied for the treatment of scarred vocal fold lamina propria due, in part, to their tunable mechanical properties, resistance to fibroblast-mediated contraction, and ability to be polymerized in situ. However, pure PEG gels lack intrinsic biochemical signals to guide cell behavior and generally fail to mimic the frequency-dependent viscoelastic response critical to normal superficial lamina propria function. Recent results suggest that incorporation of viscoelastic bioactive substances, such as glycosaminoglycans (GAGs), into PEG networks may allow these gels to more closely approach the mechanical responses of normal vocal fold lamina propria while also stimulating desired vocal fold fibroblast behaviors. Although a number of vocal fold studies have examined the influence of hyaluronan (HA) on implant mechanics and vocal fold fibroblast responses, the effects of other GAG types have been relatively unexplored. This is significant, since recent studies have suggested that chondroitin sulfate C (CSC) and heparan sulfate (HS) are substantially altered in scarred lamina propria. The present study was therefore designed to evaluate the effects of CSC and HS incorporation on the mechanical response of PEG gels and vocal fold fibroblast behavior relative to HA. As with PEG-HA, the viscoelasticity of PEG-CSC and PEG-HS gels more closely approached that of the normal vocal fold lamina propria than pure PEG hydrogels. In addition, collagen I deposition and fibronectin production were significantly higher in CSC than in HA gels, and levels of the myofibroblast marker smooth muscle α-actin (SM α-actin) were greater in CSC and HS gels than in HA gels. Since collagen I, fibronectin, and SM α-actin are generally elevated in scarred lamina propria these results suggest that CSC and HS may be undesirable for vocal fold implants relative to HA. Investigation of various signaling intermediates indicated that alterations in NFκB-p50, NFκB-p65, or pERK1/2 levels may underlie the observed differences among the PEG-GAG gels.