Genetic and genomic systems to study methylmalonic acidemia.
Methylmalonic acidemia (MMAemia) is the biochemical hallmark of a group of genetic metabolic disorders that share a common defect in the ability to convert methylmalonyl-CoA into succinyl-CoA. This disorder is due to either a mutant methylmalonyl-CoA mutase apoenzyme or impaired synthesis of adenosylcobalamin, the cofactor for this enzyme. In this article, we will provide an overview of the pathways disrupted in these disorders, discuss the known metabolic blocks with a particular focus on molecular genetics, and review the use of selected model organisms to study features of methylmalonic acidemia.


A unified approach to dose-effect relationships in radiotherapy. I: Modified TDF and linear quadratic equations.
A linear quadratic factor analogue (LQF) to the variable-exponent TDF model is introduced. In both of these models, account is taken of the volume of tissue irradiated. Scaling factors are used such that an LQF or a TDF of 100 represents tolerance for each volume or partial volume of each tissue or organ irradiated. These models are sufficient for tissues which are irradiated fairly homogeneously. Examples illustrate the use of these models.


Multivitamin pill aspiration leading to hemorrhagic bronchial necrosis.
This article highlights the serious but reversible effects of multivitamin pill aspiration into the lungs. If diagnosed early and treated promptly the harmful effects of the pill constituents on the lungs and airways can be attenuated. An early bronchoscopy and a high index of suspicion is required in order not to miss these events. We have described a case of multivitamin pill aspiration in a young healthy male and the good response to early intervention.


Assessment of Hemodynamic Conditions in the Aorta Following Root Replacement with Composite Valve-Conduit Graft.
This paper presents the analysis of detailed hemodynamics in the aortas of four patients following replacement with a composite bio-prosthetic valve-conduit. Magnetic resonance image-based computational models were set up for each patient with boundary conditions comprising subject-specific three-dimensional inflow velocity profiles at the aortic root and central pressure waveform at the model outlet. Two normal subjects were also included for comparison. The purpose of the study was to investigate the effects of the valve-conduit on flow in the proximal and distal aorta. The results suggested that following the composite valve-conduit implantation, the vortical flow structure and hemodynamic parameters in the aorta were altered, with slightly reduced helical flow index, elevated wall shear stress and higher non-uniformity in wall shear compared to normal aortas. Inter-individual analysis revealed different hemodynamic conditions among the patients depending on the conduit configuration in the ascending aorta, which is a key factor in determining post-operative aortic flow. Introducing a natural curvature in the conduit to create a smooth transition between the conduit and native aorta may help prevent the occurrence of retrograde and recirculating flow in the aortic arch, which is particularly important when a large portion or the entire ascending aorta needs to be replaced.


Lipopolysaccharide induces rapid production of IL-10 by monocytes in the presence of apoptotic neutrophils.
There is growing evidence that apoptotic neutrophils have an active role to play in the regulation and resolution of inflammation following phagocytosis by macrophages and dendritic cells. However, their influence on activated blood monocytes, freshly recruited to sites of inflammation, has not been defined. In this work, we examined the effect of apoptotic neutrophils on cytokine production by LPS-activated monocytes. Monocytes stimulated with LPS in the presence of apoptotic neutrophils for 18 h elicited an immunosuppressive cytokine response, with enhanced IL-10 and TGF-beta production and only minimal TNF-alpha and IL-1beta cytokine production. Time-kinetic studies demonstrated that IL-10 production was markedly accelerated in the presence of apoptotic neutrophils, whereas there was a sustained reduction in the production of TNF-alpha and IL-1beta. This suppression of proinflammatory production was not reversible by depletion of IL-10 or TGF-beta or by addition of exogenous IFN-gamma. It was demonstrated, using Transwell experiments, that monocyte-apoptotic cell contact was required for induction of the immunosuppressive monocyte response. The response of monocytes contrasted with that of human monocyte-derived macrophages in which there was a reduction in IL-10 production. We conclude from these data that interaction between activated monocytes and apoptotic neutrophils creates a unique response, which changes an activated monocyte from being a promoter of the inflammatory cascade into a cell primed to deactivate itself and other cells.


Two-component system regulon plasticity in bacteria: a concept emerging from phenotypic analysis of Yersinia pseudotuberculosis response regulator mutants.
In bacteria, the most rapid and efficient means of adapting gene transcription to extracellular stresses often involves sophisticated systems referred to as two-component systems (2CSs). Although highly conserved throughout the bacterial world, some of these systems may control distinct cell events and have differing contributions to virulence, depending on the species considered. This chapter summarizes the work performed by our group--from the initial PhoP-PhoQ and PmrA-PmrB studies to the most recent genome-scale preliminary analyses--in an attempt to highlight the contribution of 2CS regulon plasticity to the acquisition of some of Yersinia pseudotuberculosis' specific features.


The 3-second auditory conditioned stimulus is a more effective stressor than the 20-second auditory conditioned stimulus in male rats.
Using fear-conditioning model, we have used a 3-s auditory conditioned stimulus (CS) as a stressor and observed fear and stress responses during a specific experimental period regardless of the presence or absence of the CS. Because the CS was extremely short compared with the experimental period, we observed responses primarily in the absence of the CS. In contrast, most studies in the literature have analyzed responses in the presence of the CS. Therefore, the characteristics of fear and stress responses in the absence of the CS remain to be clarified. To clarify this, we compared the characteristics of fear and stress responses elicited by a 3-s auditory CS with those observed during a 20-s auditory CS. The basolateral complex of the amygdala (BLA), but not the bed nucleus of the stria terminalis (BNST), participated in the fear response elicited by the 3-s CS, whereas both the BLA and BNST were involved in the response observed during the 20-s CS. Additional analyses revealed that the BNST participated in the fear response during the 20-s CS when the CS was paired with a 0.75-mA, but not with a 0.9-mA, foot shock, and to the contextual CS. In addition, the fear response elicited by the 3-s CS was more resistant to extinction than that during the 20-s CS. Finally, the 3-s CS produced more intense freezing and corticosterone secretion than the 20-s CS. On the basis of these characteristics, we conclude that the 3-s auditory CS is a more effective stressor than the 20-s auditory CS. Our findings also suggest that foot shock intensity is an additional determinant in the type of fear response induced by the CS.


The metabolism of dopamine in the median eminence reflects the activity of tuberoinfundibular neurons.
The purpose of the present study was to characterize the metabolism of dopamine (DA) in tuberoinfundibular (TI) neurons terminating in the median eminence and to examine the effects of procedures that alter the synthesis and turnover of DA in these neurons on the concentrations of dihydroxyphenylacetic acid (DOPAC) in the median eminence. The DA uptake inhibitor nomifensine (25 mg/kg, i.p.; 30 min) failed to alter median eminence DOPAC concentrations indicating that very little released DA is recaptured and metabolized by TIDA neurons. Within 5 min following the administration of the monoamine oxidase inhibitor pargyline (50 mg/kg, i.v.) median eminence DOPAC concentrations declined to 15% of control demonstrating that this metabolite has a high turnover rate and is rapidly removed from the median eminence. Median eminence DOPAC concentrations in diestrous female rats, whose TIDA neuronal activity is higher than in the male, were two-fold greater than in male rats. Prolactin (10 micrograms/rat, i.c.v.; 12 h), which increases TIDA neuronal activity, produced a corresponding increase in median eminence DOPAC concentrations in male rats. Restraint stress (30 min), which decreases TIDA neuronal activity, produced a corresponding decrease in median eminence DOPAC concentrations in diestrous female rats. The results from the present study suggest that DOPAC concentrations in the median eminence can be used as an index of TIDA neuronal activity.


Neuroprotection of rhGLP-1 in diabetic rats with cerebral ischemia/reperfusion injury via regulation of oxidative stress, EAAT2, and apoptosis.
Preclinical Research & Development The purpose of the present study is to evaluate the neuroprotective effect of recombinant human glucagon-like peptide-1 (rhGLP-1) as well as to explore corresponding mechanisms in diabetic rats with cerebral ischemia/reperfusion injury induced by middle cerebral artery occlusion (MCAO). Diabetes mellitus was induced by intraperitoneal injection of streptozotocin. The rats were pretreated with rhGLP-1 (20 μg/kg intraperitoneally, thrice a day) for 14 days. Thereafter, the rats were subjected to MCAO 90 min/reperfusion 24 hr. At 2 and 24 hr of reperfusion, the rats were assessed for neurological deficits and subsequently executed for the evaluation of cerebral infarct volume, oxidative stress parameters, and the expression of excitatory amino acid transporter 2 (EAAT2) and apoptotic markers. Results indicate that rhGLP-1 significantly ameliorated neurological deficits and reduced cerebral infarct volume in diabetic MCAO rats. In addition, oxidative stress parameters in ischemic penumbra were significantly alleviated in rhGLP-1-pretreated diabetic MCAO rats. rhGLP-1 significantly upregulated the ratio of Bcl-2/Bax and EAAT2 expression and downregulated cleaved caspase-3 expression in ischemic penumbra of diabetic MCAO rats. Our results suggest that rhGLP-1 could significantly ameliorate neurological deficits and reduce cerebral infarct volume in diabetic MCAO rats, which may be due to the inhibition of oxidative stress and apoptosis and the promotion of EAAT2 expression.


Analgesic effectiveness of celecoxib and diclofenac in patients with osteoarthritis of the hip requiring joint replacement surgery: a 12-week, multicenter, randomized, double-blind, parallel-group, double-dummy, noninferiority study.
The hip is the second most common large joint that is affected by osteoarthritis (OA), with prevalence ranging from 3% to 11% in patients aged > or = 35 years. OA is often associated with significant pain, disability, and impaired quality of life. Treatment should be tailored according to the level of pain, disability, and handicap. Pharmacologic treatment options for hip OA include acetaminophen (recommended by the European League Against Rheumatism as a first-line treatment), NSAIDs such as diclofenac, and cyclooxygenase-2-selective NSAIDs such as celecoxib. The purpose of this study was to determine whether celecoxib 200 mg QD is noninferior to diclofenac 50 mg TID in the treatment of OA of the hip. This was a 12-week, randomized, double-blind, parallel-group, double-dummy, noninferiority study conducted at 40 centers in the United Kingdom. Patients with OA flare at baseline (determined by visual analog scale [VAS] measurement of > or = 40 to < 90 mm and patient's and physician's global assessments of arthritis ratings of "poor" or "very poor") and awaiting joint replacement surgery were randomized to receive celecoxib QD or diclofenac TID. Patients were excluded if surgery was anticipated within 8 weeks. The United Kingdom National Health Service initiatives on waiting-list times caused a reduction in the number of potential patients available for participation. Therefore, the study protocol was amended such that change from baseline to week 6 (as opposed to week 12) in the patient's assessment of arthritis pain on walking, measured by VAS (0-100 mm), was the primary outcome. Primary analysis was carried out on the evaluable population (subjects with baseline and week 6 arthritis pain on walking VAS scores and no major protocol deviations). Celecoxib was declared noninferior to diclofenac if the upper limit of the 2-sided 95% CI of the treatment difference (celecoxib vs diclofenac) in the mean change from baseline in VAS did not exceed 10 mm. Tolerability was assessed by the documentation of observed and volunteered adverse events (AEs), physical examination findings, sitting blood pressure, and pulse at screening and at the end of the study (week 12 or early withdrawal). A total of 249 patients aged > or = 45 years were randomized to treatment. There were 126 patients in the celecoxib group and 123 patients in the diclofenac group. One patient in the celecoxib group did not receive any treatment and was excluded from analysis. Additionally, 54 patients in the celecoxib group and 45 patients in the diclofenac group discontinued treatment due to AEs and/or lack of treatment effectiveness. Therefore, 71 patients in the celecoxib group and 78 patients in the diclofenac group completed the study. No significant differences in demographic characteristics were observed between treatment groups. The mean (SD) age was 64.0 (9.0) years, 53.9% (76/141) of the patients were men and 46.1% (65/141) were women, and 99.3% (140/141) were white. At weeks 6 and 12, the patient's assessment of arthritis pain on walking (VAS) improved in both groups (-20.0 [23.6] mm in the celecoxib group and -35 [27.0] mm in the diclofenac group [mean treatment difference, 14.4 mm; 95% CI, 6.1 to 22.7]). However, treatment differences in change from baseline favored diclofenac at week 6 (14.4 mm; 95% CI, 6.1 to 22.7) and week 12 (12.2 mm; 95% CI, 2.2 to 22.1). A post hoc analysis, performed after unblinding due to an imbalance in the numbers of patients previously receiving NSAIDs, found a greater treatment difference at week 6 between celecoxib and diclofenac in arthritis pain, favoring diclofenac, in previous nonusers of NSAIDs (n = 49, 18.6 mm; 95% CI, 4.5 to 32.8) compared with previous NSAID users (n = 92, 9.5 mm; 95% CI, -0.4 to 19.3). Celecoxib and diclofenac were generally well tolerated. A similar proportion of patients in both treatment groups experienced AEs (all causality): 67/125 of celecoxib-treated patients (53.6%) compared with 66/123 of diclofenac-treated patients (53.7%). This study did not demonstrate noninferiority of celecoxib 200 mg QD to diclofenac 50 mg TID in treating arthritis pain in patients with OA of the hip requiring joint replacement.


Endocrinological function in schizophrenic patients under haloperidol treatment: plasma PRL, HGH and 5HT levels after L-5HTP loading.
In order to examine 5HT metabolism in the hypothalamo-pituitary (HP) system of chronic schizophrenic patients taking haloperidol for a short or long period, chronological changes of blood 5HT, PRL and HGH were measured after an oral loading dose of L-5hydroxytryptophan (L-5HTP), a precursor of 5HT. The subjects consisted of 8 male patients with chronic schizophrenia, who were divided into the following two groups. The 1st group--4 patients taking haloperidol (5.6 mg/day) for 8 months on an average (short-term treatment). The 2nd group--4 patients taking haloperidol (4.8 mg/day) for 7.5 years on an average (long-term treatment). The control group was made up of 9 healthy male volunteers. As a result, the basal level of blood 5HT in the 1st group was significantly higher than that in the control group. The blood 5HT levels in the 1st and 2nd groups showed an equally remarkable increase as compared with the control group. The basal level of plasma PRL in the 2nd group was significantly lowe than that in the control group. Moreover, in the 2nd group, an increase in the plasma PRL level after the loading was suppressed, but it showed less suppression in the 1st group. There was no significant difference in the basal HGH levels among the control, 1st and 2nd groups. After the loading, an increase in the plasma HGH was suppressed in the 2nd group, but the suppression was less in the 1st group.(ABSTRACT TRUNCATED AT 250 WORDS)


Influence of spermatozoal antibodies in the reproduction of mice.
Under in vitro and in vivo conditions, the influence of spermatozoal antibodies on isolated gametes and early embryonic stages up to implantation and into postimplantation fetal development was tested. In the in vitro experiment, a clear-cut correlation between the amount of antisera added and the reduction of fertilization rates was observed. In the in vivo experiments, female mice were immunized with homologous washed sperms. A correlation between the amount of immunization and the reduction of fertilization rate occurred. In the sperm-immunized female group, the percentage of large blastocysts showed statistically significant reduction in correlation to the amount of immunization. Spermatozoal antibodies had an effect on isolated gametes, on fertilization, and on the early development of the embryo as well as on implantation. No influence of sperm antibodies either in low or high titer sperm antisera on developmental stages occurred. No spermatozoal antibody-induced abortions occurred. However, an influence of high titer antisera on implantation was evident. (Am J Reprod Immunol. 1982; 2:225-232.)


Enzymatic synthesis of oligo- and polysaccharide fatty acid esters.
Amphiphilic oligo- and polysaccharides (e.g. polysaccharide alkyl or alkyl-aryl esters) form a new class of polymers with exceptional properties. They function as polymeric surfactants, whilst maintaining most of the properties of the starting polymeric material such as emulsifying, gelling, and film forming properties combined with partial water solubility or permeability. At present carbohydrate fatty acid esters are generally obtained by chemical methods using toxic solvents and organic and inorganic catalysts that leave residual traces in the final products. Enzymatic reactions offer an attractive alternative route for the synthesis of polysaccharide esters. In this review the state of the art of enzymatic synthesis of oligo- and polysaccharides fatty esters has been described.


The effect of regular nursing rounds on patients' comfort and satisfaction, and violence against nurses in surgical ward.
Patients' satisfaction and comfort are known as the quality indicators of nursing care. Nowadays, violence against nurses has an increasing trend. Regular nursing rounds are one of the caring programs that help improve these indicators. This study aimed to examine the effect of regular nursing rounds on patients' comfort, satisfaction, and violence against nurses. This quasi-experimental study was conducted in two groups consisting of 100 patients and 35 nurses in a surgery ward in the northwest of Iran. The satisfaction with nursing care quality questionnaire, Kolcaba's general comfort questionnaire, and work environment violence were used for data collection. In the present study, the control and intervention groups were selected using a simple sampling method. The control group received routine care only; however, the intervention group received a regular nursing round program every 2 h from the second day of their admission for three days. The satisfaction questionnaire and comfort scale were completed on the second and fifth days of admission, and the evaluation of violence against nurses was performed from the second to the fourth day. The results were analyzed using chi-square, Fisher, independent t, and paired t tests. Before the intervention, no statistically significant difference was observed between the two groups in terms of demographic and dependent variables (p > 0/05). After the intervention, statistically significant differences were observed among the mean scores of satisfaction with nursing care (p < 0/001), comfort (p < 0.001), and violence against nurses (p = 0.041) between the two study groups, so that in the intervention group, the patients' satisfaction and comfort increased and violence against nurses reduced during the intervention period. The use of regular nursing rounds had a positive effect on the study results. Therefore, it is recommended that nurse managers design, implement, encourage, and evaluate regular nursing rounds to improve nursing care.


The reactions to child-bearing and early maternal experience of women with differing marital relationships.
The type of marital relationships of 200 women was significantly associated with the attitudes toward child-bearing and their affective reactions, as measured by the application of twelve content analysis scales to their retrospective accounts of a child-bearing year. Positive marital relationships were linked with calmer reactions. Self-oriented relationships were characterized by mutilation anxiety. In ambivalent and negative relationships early separation anxiety and hostility occurred but only the negative group continued in crisis after birth. Women with positive and ambivalent relationships expressed greater maternal warmth.


Excess Mortality Among Patients in the Veterans Affairs Health System Compared With the Overall US Population During the First Year of the COVID-19 Pandemic.
During the first year of the COVID-19 pandemic, there was a substantial increase in the rate of death in the United States. It is unclear whether those who had access to comprehensive medical care through the Department of Veterans Affairs (VA) health care system had different death rates compared with the overall US population. To quantify and compare the increase in death rates during the first year of the COVID-19 pandemic between individuals who received comprehensive medical care through the VA health care system and those in the general US population. This cohort study compared 10.9 million enrollees in the VA, including 6.8 million active users of VA health care (those with a visit in the last 2 years), with the general population of the US, with deaths occurring from January 1, 2014, to December 31, 2020. Statistical analysis was conducted from May 17, 2021, to March 15, 2023. Changes in rates of death from any cause during the COVID-19 pandemic in 2020 compared with previous years. Changes in all-cause death rates by quarter were stratified by age, sex, race and ethnicity, and region, based on individual-level data. Multilevel regression models were fit in a bayesian setting. Standardized rates were used for comparison between populations. There were 10.9 million enrollees in the VA health care system and 6.8 million active users. The demographic characteristics of the VA populations were predominantly male (>85% in the VA health care system vs 49% in the general US population), older (mean [SD], 61.0 [18.2] years in the VA health care system vs 39.0 [23.1] years in the US population), and had a larger proportion of patients who were White (73% in the VA health care system vs 61% in the US population) or Black (17% in the VA health care system vs 13% in the US population). Increases in death rates were apparent across all of the adult age groups (≥25 years) in both the VA populations and the general US population. Across all of 2020, the relative increase in death rates compared with expected values was similar for VA enrollees (risk ratio [RR], 1.20 [95% CI, 1.14-1.29]), VA active users (RR, 1.19 [95% CI, 1.14-1.26]), and the general US population (RR, 1.20 [95% CI, 1.17-1.22]). Because the prepandemic standardized mortality rates were higher in the VA populations prior to the pandemic, the absolute rates of excess mortality were higher in the VA populations. In this cohort study, a comparison of excess deaths between populations suggests that active users of the VA health system had similar relative increases in mortality compared with the general US population during the first 10 months of the COVID-19 pandemic.


Combined chemoradiotherapy in limited-disease, inoperable non-small cell lung cancer.
Forty-three patients with limited-disease, inoperable non-small cell lung cancer received two intravenous courses of cyclophosphamide, Adriamycin, and cisplatin (CAP) chemotherapy over a 6-week period. This was followed by 5 weeks of combined chemoradiotherapy (CCRT) consisting of low weekly doses of CAP for 5 weeks plus 50 Gy continuous X ray therapy (XRT) to the primary tumor site. Chemotherapy was continued until disease progression occurred or until the total dose of Adriamycin reached 450 mg/m2, whichever came first. CCRT improved the response rate [complete response (CR) plus partial responses (PR)] from 25% after two courses of CAP alone to 65% after CCRT. Previous response to two courses of CAP influences response subsequent to CAP plus XRT. A pretherapy weight loss of 6% or greater had a significant adverse effect on both response and survival time. The median survival time for all patients was 50 weeks; patients whose disease responded to treatment survived significantly longer than patients with nonresponding disease. The median time until disease progression was 37 weeks. Twenty-seven patients relapsed. The first sites of relapse were local in 30% of the patients, distant in 56% of them, and both local and distant in 15%. Severe esophagitis occurred in 30% of the patients and was dose-limiting. The administration of CCRT resulted in an improved response rate compared with the rates reported in previous studies of chemotherapy or radiotherapy alone. Further improvement of the CCRT program is needed to increase long-term survival time and to decrease esophageal toxicity.


Effects of dietary administration of water hyacinth (Eichhornia crassipes) extracts on the immune responses and disease resistance of giant freshwater prawn, Macrobrachium rosenbergii.
The hot-water extract of Eichhornia crassipes leaves (ECE) was produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Survival rates of prawn against Lactococcus garvieae, and its immune parameters including the total haemocyte count (THC), different haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, transglutaminase (TG) activity, haemolymph coagulation time, and phagocytic activity and clearance efficiency against L. garvieae by M. rosenbergii were determined when prawn (23.0 ± 2.8 g) were fed ECE-containing diets at 0, 1.0, 2.0 and 3.0 g kg(-1). Prawn fed a diet containing ECE at 2.0 and 3.0 g kg(-1) for 12 days showed significantly increased THC, HC, GC, PO activity, RBs, SOD activity, GPx activity, and TG activity, and a significantly decreased coagulation time. The phagocytic activity and clearance efficiency against L. garvieae of prawn fed the ECE-containing diets at 1.0, 2.0, and 3.0 g kg(-1) were significantly higher than those of prawn fed the control diet at 3-12 days. Survival rates of M. rosenbergii fed the diet containing ECE at concentrations of 2 and 3 g kg(-1) were significantly higher than those fed the control diet after challenge with L. garvieae for 48-144 h. The relative percentage survival of prawn fed the 1.0, 2.0, and 3.0 g kg(-1) ECE-containing diets for 12 days were 17.5%, 39.1%, and 52.2%. It was concluded that the ECE can be used as an immunostimulant for prawn through dietary administration to enhance immune responses and resistance of M. rosenbergii against L. garvieae.


Investigation of Circulating MicroRNA Levels in Antibody-Mediated Rejection After Kidney Transplantation.
One of the most important possible complications determining long-term graft survival after kidney transplant is antibody-mediated rejection (ABMR). The criterion standard approach to recognize ABMR is currently the kidney biopsy with histopathologic analysis. However, this test has limitations because of difficulties in timing of sampling, the evaluability of histology because of the questionable representativeness of specimens, and the limited number of this intervention. Hence, new reliable, noninvasive biomarkers are required to detect the development of ABMR in time. In this study, we analyzed the clinical data of 45 kidney transplant patients (mean age of 44.51 years, 20 male and 25 female subjects). These participants were recruited into 5 subcohorts based on their clinical status, histologic findings, and level of donor-specific anti-HLA antibodies. Circulating microRNAs (miR-21, miR-181b, miR-146a, miR-223, miR-155, miR-150) in plasma samples were quantified by quantitative polymerase chain reaction and their levels were correlated with the clinical characteristics in different subgroups. The relative expression of plasma miR-155 (P = .0003), miR-223 (P = .0316), and miR-21 (P = .0147) were significantly higher in patients who had subsequent histology-approved ABMR with donor-specific anti-HLA antibody positivity (n = 10) than in the "triple negative" group (n = 21), and miR-155 showed the highest sensitivity (90%) and specificity (81%) to indicate ABMR development based on receiver operating characteristic analysis. According to our preliminary data, plasma miR-155, miR-21, and miR-223 can indicate the development of ABMR after kidney transplant in correlation with classic clinical parameters. However, future studies with larger number of participants are necessary to further evaluate the diagnostic properties of blood miRNAs in prediction of this life-threatening condition.


Characterization of a Glycoside Hydrolase Family 27 α-Galactosidase from Pontibacter Reveals Its Novel Salt-Protease Tolerance and Transglycosylation Activity.
α-Galactosidases are of great interest in various applications. A glycoside hydrolase family 27 α-galactosidase was cloned from Pontibacter sp. harbored in a saline soil and expressed in Escherichia coli. The purified recombinant enzyme (rAgaAHJ8) was little or not affected by 3.5-30.0% (w/v) NaCl, 10.0-100.0 mM Pb(CH3COO)2, 10.0-60.0 mM ZnSO4, or 8.3-100.0 mg mL(-1) trypsin and by most metal ions and chemical reagents at 1.0 and 10.0 mM concentrations. The degree of synergy on enzymatic degradation of locust bean gum and guar gum by an endomannanase and rAgaAHJ8 was 1.22-1.54. In the presence of trypsin, the amount of reducing sugars released from soybean milk treated by rAgaAHJ8 was approximately 3.8-fold compared with that treated by a commercial α-galactosidase. rAgaAHJ8 showed transglycosylation activity when using sucrose, raffinose, and 3-methyl-1-butanol as the acceptors. Furthermore, potential factors for salt adaptation of the enzyme were presumed.


Statistical consequences of a successful lung allocation system - recovering information and reducing bias in models for urgency.
The lung allocation system has reduced the number of waitlist deaths by ranking transplant candidates on the basis of a lung allocation score that requires estimation of the current 1-year restricted mean waitlist survival (urgency). Fewer waitlist deaths and the systematic removal of candidates from the waitlist for transplantation present statistical challenges that must be addressed when using recent waitlist data. Multiple overlapping 1-year follow-up windows are used in a restricted mean model that estimates patient urgency on the basis of updated risk factors at the start of the window. In simulation studies, our proposed multiple imputation procedure was able to produce unbiased parameter estimates with similar efficiency to those obtained if censoring had never occurred. The analysis of 10,740 lung transplant candidates revealed that for most risk factors incorporating additional follow-up windows produced more efficient estimates. Copyright © 2017 John Wiley & Sons, Ltd.


Predicting soil available cadmium by machine learning based on soil properties.
Cadmium (Cd) accumulation in edible plant tissues poses a serious threat to human health through the food chain. Assessing the availability of soil Cd is crucial for evaluating associated environmental risks. However, existing experimental methods and traditional models are time-consuming and inefficient. In this study, we developed machine learning models to predict soil available Cd based on soil properties, using a dataset comprising 585 data points covering 585 soils. Traditional machine learning models exhibited prediction values beyond the theoretical range, urging the need for alternative approaches. To address this, different models were tested, and the post-constraint eXtreme Gradient Boosting (XGBoost) model was found to possess the best predictive performance (R2 =0.81) outperform traditional linear regression model in terms of accuracy. Furthermore, we explored the relationship between soil available Cd and wheat grain Cd and rice grain Cd. Linear regression models were developed using 302 data points for wheat and 563 data points for rice. Results demonstrated a significant correlation between soil available Cd and wheat grain Cd (R2 =0.487) as well as rice grain Cd (R2 =0.43).


Prescribing patterns of antilipemic drugs and prevalence of hypercholesterolemia in the Nova Scotia population more than 65 years old.
To examine the prevalence of antilipemic drug use, demographic characteristics of patients using these drugs, and the prevalence of hypercholesterolemia in the Nova Scotia population over 65 years of age. Information was collected on the prescribing of antilipemic drugs using Nova Scotia Medical Services Insurance Pharmacare program data from October 1991 through March 1992. Pharmacare data were compared with prevalence data on increased low-density lipoprotein (LDL) cholesterol concentrations obtained from the Nova Scotia Heart Health Survey (NSHHS). Pharmacare is a centrally administered drug insurance system maintained in computerized claims files since 1974. It provides prescription drugs to all residents of Nova Scotia who are at least 65 years old and who are insured under the provincial Medicare program. In the 1991-1992 fiscal year, 47,000 men and 65,700 women were eligible for Pharmacare. The NSHHS was administered to a probability sample of 2,108 individuals, representative of the 1986 population aged 18-74 years. Prescriptions for antilipemic agents. The NSHHS data indicated that 3.7% of women and 2.3% of men at least 65 years old and 4.8% of women and 2.8% of men 65-74 years old received a prescription for antilipemic drugs.


Contributions of mechanical and sonochemical effects to cell membrane damage induced by single-shot pulsed ultrasound with adjacent microbubbles.
The objective was to investigate the contributions of mechanical effects due to kinetic force induced by the dynamic behavior of microbubbles and sonochemical effects due to free radicals produced by inertial cavitation to cell membrane damage under sonoporation conditions in which cells with adjacent microbubbles were irradiated with single-shot pulsed ultrasound. The free radical scavenger cysteamine was used to control the occurrence of sonochemical effects, and the ratios of cells with membrane damage to intact cells were compared in the presence and absence of cysteamine. To determine the optimal dose of cysteamine, free radical production on exposure to burst pulse ultrasound was investigated using KI-starch solutions with different concentrations (0-5 mM) of cysteamine. High-speed observation of the dynamic behavior of Levovist microbubbles during ultrasound exposure was also carried out in the presence and absence of cysteamine, and the difference in the ratios of the maximum bubble diameter to the initial diameter was evaluated. Next, human prostate cancer cells with adjacent Levovist microbubbles were exposed to single-shot pulsed ultrasound with a center frequency of 1 MHz, a peak negative pressure of 1.1 MPa, and a pulse width of 3 μs, and the percentages of cells with membrane damage were evaluated by fluorescent microscopy using propidium iodide in the presence and absence of cysteamine. It was confirmed that cysteamine at a concentration of 5 mM completely suppressed sonochemical effects without causing a change in the dynamic response of microbubbles to pulsed ultrasound. The percentages of cells with membrane damage in the presence and absence of cysteamine (5 mM) were 10.3% ± 4.1% (n = 13) and 8.7% ± 3.9% (n = 9), respectively. No significant difference was found (P = 0.36). The results indicate that cell membrane damage induced by single-shot pulsed ultrasound with adjacent microbubbles was due mainly to mechanical effects, not to sonochemical effects.


Idazoxan blocks the nicotine-induced reversal of the memory impairment caused by the NMDA glutamate receptor antagonist dizocilpine.
Alpha2-adrenoreceptor (alpha(2)-AR) antagonists have been shown to improve, while alpha(2)-AR agonists impair cognitive function in subjects with functioning NMDA receptors (NMDAR). In subjects with inhibited NMDAR (a model of schizophrenia) alpha(2)-AR agonists attenuate the cognitive impairments. The effect with alpha(2)-AR antagonists remains unclear. We investigated the effects of the alpha(2)-AR antagonist idazoxan on memory function in rats treated/not treated with NMDAR antagonist dizocilpine or a combination of dizocilpine and nicotine to clarify noradrenergic/cholinergic regulation of memory function. Female Sprague-Dawley rats (n=12) were trained for food reward on the radial maze. Working and reference memory errors and response latency were assessed after injections of idazoxan (0.5, 1.0 mg/kg), dizocilpine (0.05 mg/kg), nicotine (0.2, 0.4 mg/kg) or vehicle, alone or in combination. Dizocilpine potently impaired memory. Nicotine (0.4 mg/kg) reversed this impairment. Idazoxan at the doses tested did not affect performance when given alone or with dizocilpine, but it did block the nicotine reversal of the dizocilpine-induced memory impairment. Three rats after 10-12 drug treatments developed limbic seizures. Our findings suggest that combination of drugs which block alpha(2)-AR with nicotinic agonists in schizophrenia may prevent therapeutic effect of nicotinic agonists and increase risk for convulsive activity with repeated administration.


Rates, Causes, and Reduction of 30-Day Readmissions of Otolaryngology-Head and Neck Surgical Cases.
The aim of this study was to determine risk factors associated with 30-day readmission for patients undergoing inpatient otolaryngologic head and neck surgery. Retrospective cohort study analysis. Study at 2 tertiary hospitals. A 10-year retrospective cohort analysis was performed for 30-day readmissions of otolaryngology surgical cases between July 1, 2006, and June 30, 2016, at Assir Central Hospital and Abha Private Hospital. Data included total number of patients, type of surgical procedure, number of and reasons for readmissions, and length of hospital stay. There were 32,662 discharges for otolaryngology operations over the 10-year period of the study, of which 364 patients were readmitted, giving a rate of 11.14 readmissions per 1000 operative procedures (95% CI, 10.1-12.3). The male:female ratio was 1.4:1. Period of postoperative stay ranged from 1 to 3 days and, after readmission, 2 to 5 days. The main reasons for readmission were bleeding in otolaryngologic cases and wound hematoma in head and neck surgical cases. Overall readmission rates dropped significantly from 12.72 per 1000 operative procedures in the first 5 years to 10.16 in the second 5 years. This study helped to establish special policies and procedures to prevent readmission by utilizing best practices, including addressing quality care, using preadmission clinics, preventing surgical site infection, and improving communication with community physicians. Plans based on these results also include the development of national model for predicting readmission within 30 days of discharge.


Ternary complex formation of pig heart lactate dehydrogenase with spin-labelled coenzyme and inhibitors as studied by electron spin resonance.
The formation of ternary inhibitor and 'dead end' complexes of pig heart lactate dehydrogenase (L-lactate:NAD+ oxidoreductase, EC 18.104.22.168) was studied by means of two NAD derivatives, spin-labelled at N6 and C-8 of the adenine ring. Dissociation constants calculated for the inhibitors oxamate and oxalate from their corresponding ternary complexes are in excellent agreement with data from literature derived from sedimentation experiments. However, the recently postulated enzyme-NADH-sulfite complex was not observed. The mobility of the spin-label, i.e. the protein conformation near the adenine binding pocket in various ternary complexes depends on the type of inhibition or substrate employed.


Polymorphism of glutathione S-transferase P1 gene affects human vitamin C metabolism.
There are large inter-individual differences in the metabolism of vitamin C (VC), which is composed of both ascorbic acid (AsA) and dehydroascorbic acid (DAsA). AsA is oxidized to DAsA in a series of xenobiotic reactions. Thus, the effects of polymorphism A313G (Ile105Val) in the gene for glutathione S-transferases P1 (GSTP1), one of the most active xenobiotic enzymes, on human VC metabolism were studied. The variant frequency of GSTP1 among the present subjects (n=210) was AA 71.0%; GA 27.0% and GG 1.9%. At 24 h after administration of 1 mmol of VC to young women (n=17; age, 21.0+/-1.1 y), total VC excretion (46.7+/-18.1mg) by AA homozygotes of GSTP1 was greater (p<0.0069) than that (28.2+/-14.0 mg) by GA heterozygotes. One hour after administration of VC, blood total VC levels were also significantly different (p<0.0036) between the homozygotes and heterozygotes. The effects of other polymorphisms in xenobiotic enzymes on VC metabolism were small.


Characteristics and prognostic importance of ST-segment elevation on Holter monitoring early after acute myocardial infarction.
The correlation between episodes of ST-segment elevation on Holter monitoring, clinical characteristics, left ventricular function, exercise testing, and long-term prognosis was determined in 123 consecutive patients 55 +/- 8 years old (mean +/- SD) with a first acute myocardial infarction (AMI). During 36 hours of Holter recording 11 +/- 5 days after AMI, 11 patients (9%) had 91 episodes of ST-segment elevation (group 1), whereas 112 patients had no such episodes (group 2). Most episodes of ST-segment elevation occurred in leads with pathologic Q waves or small, indistinct R waves. Large, anterior Q-wave AMIs were more prevalent in group 1 than in group 2, and in-hospital heart failure also occurred more frequently in group 1 patients (82% vs 23%; p < 0.0005). Regional and global left ventricular function was reduced in group 1 compared with group 2: ejection fraction 33 +/- 11% vs 50 +/- 11% (p = 0.0001). All episodes of ST-segment elevation were asymptomatic and did not correlate with different indicators of myocardial ischemia. Indeed, exercise-induced ST-segment depression was more prevalent in group 2 than in group 1: 57 vs 18% (p < 0.035). Over a mean of 5 years (range 4 to 6) of follow-up, an association between episodes of ST-segment elevation on Holter monitoring and (1) cardiac death (Kaplan-Meier analysis; p < 0.005), and (2) cardiac death and nonfatal reinfarction (Kaplan-Meier analysis; p < 0.025) was found.(ABSTRACT TRUNCATED AT 250 WORDS)


Low turnover osteoporosis in sheep induced by hypothalamic-pituitary disconnection.
The hypothalamus is of critical importance in regulating bone remodeling. This is underscored by the fact that intracerebroventricular-application of leptin in ewe leads to osteopenia. As a large animal model of osteoporosis, this approach has some limitations, such as high technical expenditure and running costs. Therefore we asked if a surgical ablation of the leptin signaling axis would have the same effects and would thereby be a more useful model. We analyzed the bone phenotype of ewe after surgical hypothalamo-pituitary disconnection (HPD + OVX) as compared to control ewe (OVX) after 3 and 12 months. Analyses included histomorphometric characterization, micro-CT and measurement of bone turnover parameters. Already 3 months after HPD we found osteopenic ewe with a significantly decreased bone formation (69%) and osteoclast activity (49%). After a period of 12 months the HPD group additionally developed an (preclinical) osteoporosis with significant reduction (33%) of femoral cortical thickness, as compared to controls (OVX). Taken together, HPD leads after 12 month to osteoporosis with a reduction in both trabecular and cortical bone caused by a low bone turnover situation, with reduced osteoblast and osteoclast activity, as compared to controls (OVX). The HPD-sheep is a suitable large animal model of osteoporosis. Furthermore our results indicate that an intact hypothalamo-pituitary axis is required for activation of bone turnover.


PFK2, ISP42, ERG2 and RAD14 are located on the right arm of chromosome XIII.
An 11 kb yeast DNA insertion isolated from a genomic library by complementation of a phosphofructokinase-deficient strain was used as a source for a partial sequence analysis. Four genes were shown to reside on this fragment, namely PFK2, ISP42, ERG2 and RAD14. PFK2 was mapped previously to the right arm of chromosome XIII, locating the latter three genes to the same chromosome.


Isolation and identification of the broad-spectrum high-efficiency phage vB_SalP_LDW16 and its therapeutic application in chickens.
Salmonella infection in livestock and poultry causes salmonellosis, and is mainly treated using antibiotics. However, the misuse use of antibiotics often triggers the emergence of multi-drug-resistant Salmonella strains. Currently, Salmonella phages is safe and effective against Salmonella, serving as the best drug of choice. This study involved 16 Salmonella bacteriophages separated and purified from the sewage and the feces of the broiler farm. A phage, vB_SalP_LDW16, was selected based on the phage host range test. The phage vB_SalP_LDW16 was characterized by the double-layer plate method and transmission electron microscopy. Furthermore, the clinical therapeutic effect of phage vB_SalP_LDW16 was verified by using the pathogenic Salmonella Enteritidis in the SPF chicken model. The phage vB_SalP_LDW16 with a wide host range was identified to the family Siphoviridae and the order Caudoviridae, possess a double-stranded DNA and can lyse 88% (22/25) of Salmonella strains stored in the laboratory. Analysis of the biological characteristics, in addition, revealed the optimal multiplicity of infection (MOI) of vB_SalP_LDW16 to be 0.01 and the phage titer to be up to 3 × 1014 PFU/mL. Meanwhile, the phage vB_SalP_LDW16 was found to have some temperature tolerance, while the titer decreases rapidly above 60 ℃, and a wide pH (i.e., 5-12) range as well as relative stability in pH tolerance. The latent period of phage was 10 min, the burst period was 60 min, and the burst size was 110 PFU/cell. Furthermore, gastric juice was also found to highly influence the activity of the phage. The clinical treatment experiments showed that phage vB_SalP_LDW16 was able to significantly reduce the bacterial load in the blood through phage treatment, thereby improving the pathological changes in the intestinal, liver, and heart damage, and promoting the growth and development of the chicken. The phage vB_SalP_LDW16 is a highly lytic phage with a wide host range, which can be potentially used for preventing and treating chicken salmonellosis, as an alternative or complementary antibiotic treatment in livestock farming.


Geographical inequality in service utilization for primary aldosteronism screening: spatial epidemiological study in Southern Thailand.
Primary aldosteronism (PA), the most common cause of secondary hypertension is considered as a "major public health issue" due to higher risk of cardiovascular complications compared to blood-pressure-match hypertension and increase in prevalence around the world. In Thailand, though PA screening is provided under the universal health coverage, the service can be offered only at some centers. Hence, the service availability affects an accessibility of health care in patients. Our study aimed to evaluate the service utilization in PA screening and diagnosis in terms of geographical inequality in health resources in Southern Thailand. Data of 688 patients who underwent PA screening from 2011 to 2017 were obtained from the electronic database of Songklanagarind Hospital, a super-tertiary center in this region. The patients' residence in the province, district and subdistrict were transformed to a 6-digit numbers corresponding to the global one (GADM©). The areas with PA screening and diagnosis were visualized by disease mapping procedures. A general log linear model was used to identify the factors affecting patient's service accessibility. From the geographic distribution, patients living in or near the area of the super-tertiary center (Songkhla) had high probability of receiving PA screening. The analysis of factors contributing to PA screening by multivariate log-linear model demonstrated that the distance from the super-tertiary center was a predictive factor for screening while the presence of endocrinologists and cultural differences were not. The chance of patients living in Songkhla, living less than 200 km, and more than 200 km from Songkhla to receive PA screening was 100, 82, and 66%, respectively. The crude incidence rate of PA in Southern Thailand was 1.66/106 person-years. The provinces located adjacent to the Andaman Sea had the highest incidences of PA (3.62-5.17 patients/106 person-years). There is still geographical inequality and the strategy to decrease the barrier should be resolved. The policymaker should develop a transfer system of blood tests for PA investigation from the local hospital to reduce the burden such as transportation costs in patients who live far away from the super-tertiary hospital. In addition, PA screening should be implemented in hypertension care plan.


The precipitation of proteins by carboxymethyl cellulose.
Carboxymethyl cellulose (CMC) formed insoluble complexes with beta-lactoglobulin, bovine serum albumin and Na caseinate. Maximum precipitation of the beta-lactoglobulin-CMC complex occurred at pH 3-2, whereas maximum precipitation of the bovine serum albumin. The percentage of protein precipitated by CMC decreased with increasing ionic strength of the solution, the rate of decrease being least for bovine serum albumin. At a given ionic strength, more protein was precipitated by CMC of high degree of substitution than by CMC of low degree of substitution. The change in pH (delta pH) occurring on mixing CMC and unbuffered protein solutions, each initially at the same pH, was measured. delta pH was negative for beta-lactoglobulin-CMC mixtures over the pH range 7--2 (minimum at pH 5-5). For bovine serum albumin--Cmc and Na caseinate--CMC mixtures, delta pH was positive between pH 7 and 3-2 (maximum at pH 4-5), zero at pH 3-2 and negative between pH 3-2 and 2-0 (minimum at pH 2-8).


Human temporal-parietal junction spontaneously tracks others' beliefs: A functional near-infrared spectroscopy study.
Humans have the unique capacity to actively reflect on the thoughts, beliefs, and knowledge of others, but do we also track mental states spontaneously when observing other people? We asked this question by monitoring brain activity in belief-sensitive cortex using functional near-infrared spectroscopy (fNIRS) during free-viewing of social videos. More specifically, we identified a portion of the right temporal-parietal junction (rTPJ) selective for mental state processing using an established, explicit theory of mind task, and then analyzed the brain response in that region of interest (ROI) during free-viewing of video clips involving people producing goal-directed actions. We found a significant increase in oxygenated hemoglobin concentration in our rTPJ ROI during free-viewing for all of our test videos. Activity in this region was further modulated by the extent to which the knowledge state, or beliefs, of the protagonist regarding the location of an object contrasted with the reality of where the object was hidden. Open-ended questioning suggested our participants were not explicitly focusing on belief states of the characters during free-viewing. Further analyses ruled out lower-level details of the video clips or general attentional differences between conditions as likely explanations for the results. As such, these results call into question the traditional characterization of theory of mind as a resource intensive, deliberate process, and, instead, support an emerging view of theory of mind as a foundation for, rather than the pinnacle of, human social cognition.


IGF1 and PPARG polymorphisms are associated with reduced estimated glomerular filtration rate in a cohort of children and adolescents with type 1 diabetes.
Several genetic loci have been associated with diabetic nephropathy; however, the underlying genetic mechanisms are still poorly understood, with no robust candidate genes identified yet. We aimed to determine whether two polymorphisms, previously associated with renal decline, influence kidney impairment evaluating their association with markers of renal function in a pediatric population with type 1 diabetes (T1D). Renal function was evaluated by glomerular filtration rate (eGFR) and albumin-to-creatinine ratio (ACR) in a cohort of pediatric subjects with T1D (n = 278). Risk factors for diabetes complications (diabetes duration, blood pressure, HbA1c) were assessed. The IGF1 rs35767 and PPARG rs1801282 SNPs were genotyped by TaqMan RT-PCR system. An additive genetic interaction was calculated. Association analysis between markers of renal function and both SNPs or their additive interaction were performed. Both SNPs showed a significant association with eGFR: the A allele of rs35767 or the C allele of rs1801282 were associated to reduced eGFR compared to G alleles. Multivariate regression analysis adjusted for age, sex, z-BMI, T1D duration, blood pressure and Hba1c values showed that the additive genetic interaction was independently associated with lower eGFR (β = -3.59 [-6.52 to -0.66], p = 0.017). No associations were detected between SNPs, their additive interaction and ACR. These results provide new insight into the genetic predisposition to renal dysfunction, showing that two polymorphisms in IGF1 and PPARG genes can lead to a reduction in renal filtration rate leading these patients to be exposed to a higher risk of early renal complications.


Machine Learning from Omics Data.
Machine learning (ML) already accelerates discoveries in many scientific fields and is the driver behind several new products. Recently, growing sample sizes enabled the use of ML approaches in larger omics studies. This work provides a guide through a typical analysis of an omics dataset using ML. As an example, this chapter demonstrates how to build a model predicting Drug-Induced Liver Injury based on transcriptomics data contained in the LINCS L1000 dataset. Each section covers best practices and pitfalls starting from data exploration and model training including hyperparameter search to validation and analysis of the final model. The code to reproduce the results is available at https://github.com/Evotec-Bioinformatics/ml-from-omics .


Does a contralateral carotid occlusion adversely impact carotid artery stenting outcomes?
Carotid artery stenting (CAS) has grown as a possible alternative for the treatment of extracranial cerebrovascular disease in the past decade. A preexisting contralateral carotid artery occlusion has been described as a risk factor for inferior outcomes after carotid endarterectomy, but its impact on CAS outcomes is less understood. A retrospective review of 417 CAS procedures performed between May 2001 and July 2010 at a single center using self-expanding nitinol stents and mechanical embolic protection devices was conducted. Patients were divided into two groups, those with a preexisting contralateral carotid occlusion (group A, n = 39) versus those without a contralateral occlusion (group B, n = 378). Patient demographics and comorbidities as well as 30-day and late death, stroke, and myocardial infarction (MI) rates were analyzed. Mean follow-up was 4 years (range: 0-9.4 years). Overall, mean age of the 314 men and 103 women was 70.5 years. In group A, there were two (5.1%) octogenarians and nine patients (23.1%) with symptomatic disease as compared with group B with 53 (14%) octogenarians and 121 (32%) patients with symptomatic disease. The overall 30-day death, stroke, and MI rates were 0.5%, 1.9%, and 0.7%, respectively. When comparing group A with group B, these results were not significantly different: death (0% vs. 0.5%), stroke (2.6% vs. 1.9%), and MI (0% vs. 0.8%). Long-term outcomes for groups A and B were also not significantly different: death (25.6% vs. 22.2%), stroke (5.3% vs. 3.4%), and MI (15.4% vs. 14%) (p = nonsignificant). A preexisting contralateral carotid artery occlusion does not seem to adversely impact CAS outcomes.


Determination of the ideal volume of activating solution and the optimal ratio of spermatozoa per oocyte for Prochilodus brevis fertilization.
Prochilodus brevis is a rheophilic species with a threatened natural population that promotes studies aimed at optimizing reproduction in captivity. The correct quantity of inseminating dose and activating solution volume significantly improves fertilization rates, thereby increasing productivity in captivity. The objective of this study was to determine the proportion of sperm per oocyte and the ideal volume of activating solution to be used in the assisted fertilization of P. brevis. Gametes were collected and fertilization performed in two steps. In step 1, the ideal proportion of spermatozoa was determined based on the fertilization rate:oocyte by testing six doses of semen: D1 = 30 × 103, D2 = 150 × 103, D3 = 300 × 103, D4 = 3 × 106, D5 = 5 × 106, and D6 = 10 × 106. In step 2, the fertilization and hatching rates were evaluated using different volumes of activating solution (V1 - 25 ml, V2 - 50 ml, V3 - 75 ml,V4 - 100 ml, V5 - 125 ml, and V6 - 150 ml). A linear regression equation was estimated from steps 1 and 2. The Student-Newman-Keuls test was used to compare the means. In step 1, the percentage of fertilization increased linearly, reaching a plateau of 51.69%. In step 2, the best fertilization rates were obtained with an estimated ideal volume of 75.64 ml per 2 ml of oocytes. Therefore, the proportion of 928,410.29 sperm:oocyte, associated with the volume of 75.64 ml of water per 2 ml of oocytes, provided the maximum reproductive performance for P. brevis.


An essential arginine residue in human prostatic acid phosphatase.
Treatment of human prostatic acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 220.127.116.11) with either of the arginine-specific modifiers 2,3-butanedione or 1,2-cyclohexanedione in borate buffer at pH 8.1 leads to loss of activity. The inactivation by cyclohexanedione can be partially reversed by 0.2 M hydroxylamine. The rate of inactivation by both modifiers is decreased in the presence of the competitive inhibitors L-(+)-tartrate or inorganic phosphate but not in the presence of the non-inhibitor D-(-)-tartrate. Amino acid analysis of modified acid phosphatase indicates that only arginines are modified and that L-(+)-tartrate protects at least two arginyl residues from modification. A likely role of these arginyl residues is their involvement in binding the negatively charged phosphate group of the substrate.


Intrafamilial transmission of Helicobacter pylori: the association between a parent and an offspring with respect to the presence of anti-CagA antibody.
In this study, we examined whether the transmission of Helicobacter pylori occurred in married couples. We also examined whether a correlation existed between H. pylori seropositivity in a parent and an offspring, which may be related to H. pylori transmission. A cross-sectional study of residents living in a rural area in Japan was conducted in 1998. The study population consisted of 1910 volunteers, aged from 22 to 79 years, residing in this area. We serologically confirmed the presence of the antigen, CagA (cytotoxin-associated gene A), of H. pylori, using a recombinant CagA antigen, in addition to examining for antibodies to H. pylori, as determined by an IgG-specific enzyme immunoassay. The data were analyzed using logistic regression models. A significant association of H. pylori seropositivity was observed ( P < 0.001) in married couples, but no significant difference with respect to the presence of anti-CagA antibodies was observed in the married couples with H. pylori seropositive results ( P = 0.053). The odds ratio was 8.08 (95% confidence interval [CI], 1.39-40.05) for infection in offspring with seropositive mothers and 2.93 (95% CI, 0.25-33.85) for infection in offspring with seropositive fathers when compared with seronegative fathers or mothers as the reference. There was a concordance between the presence or absence of anti-CagA antibodies in 11 of 13 groups of mothers and offspring (84.6%) compared with concordance in 6 of 14 groups of fathers and offspring (42.9%; P < 0.05). Our results suggest that maternal influence is likely to be more powerful than paternal influence in the transmission of H. pylori.


Metagenomic study of endophytic bacterial community of sweet potato (Ipomoea batatas) cultivated in different soil and climatic conditions.
The aim of this study was to clarify effects of soil and climatic conditions on community structure of sweet potato bacterial endophytes by applying locked nucleic acid oligonucleotide-PCR clamping technique and metagenomic analysis. For this purpose, the soil samples in three locations were transferred each other and sweet potato nursery plants from the same farm were cultivated for ca. 3 months. After removal of plastid, mitochondria and undefined sequences, the averaged numbers of retained sequences and operational taxonomic units per sample were 20,891 and 846, respectively. Proteobacteria (85.0%), Bacteroidetes (6.6%) and Actinobacteria (6.3%) were the three most dominant phyla, accounting for 97.9% of the reads, and γ-Proteobacteria (66.3%) being the most abundant. Top 10 genera represented 81.2% of the overall reads in which Pseudomonas (31.9-45.0%) being the most predominant. The overall endophytic bacterial communities were similar among the samples which indicated that the soil and the climatic conditions did not considerably affect the entire endophytic community. The original endophytic bacterial community might be kept during the cultivation period.


Effect of anticonvulsant drugs on interleukins-1, -2 and -6 and monocyte chemoattractant protein-1.
In order to evaluate whether treatment with valproic acid or carbamazepine can modify interleukins and monocyte chemoattractant protein-1, we studied 40 epileptic children and adolescents. We evaluated the patients before and after 1 year of therapy. At the end of follow-up, the patients showed a significant increase of the production of interleukin-1alpha, interleukin-1beta, interleukin-6, and monocyte chemoattractant protein-1; interleukin-2 production was significantly higher only in patients receiving carbamazepine. In conclusion, antiepileptic drugs can influence the immune system by modifying interleukin and chemokine concentrations; these changes seem to be independent of the serum concentrations of these drugs.


Vision loss in tuberculous meningitis.
Vision loss is a disabling complication of tuberculous meningitis. Approximately, 15% of survivors are either completely or partially blind. All structures of the visual pathway may be affected in tuberculous meningitis. Optic nerve and optic chiasma are most frequently and dominantly affected. Thick-gelatinous exudates lying over the base of brain, are the pathological hallmark of tuberculous meningitis and are responsible for almost all of its major complications, including vision loss. Strangulation of optic nerves and optic chiasma by the exudates, compression over optic chiasma by the dilated third ventricle, raised intracranial pressure, endarteritis, shunt failure, bacterial invasion of optic nerves and drug-induced optic nerve damage are important reasons that are considered responsible for vision loss. Prompt antituberculosis treatment is the best management option available. Immunomodulatory drugs and cerebrospinal fluid diversion procedures are of limited help. Early recognition and treatment of tuberculous meningitis is the only way forward to tackle this problem.


Nitric oxide metabolites in hypoxia, freezing, and hibernation of the wood frog, Rana sylvatica.
Nitric oxide (NO) is a gaseous free radical that in diverse organisms performs many signaling and protective functions, such as vasoregulation, inhibition of apoptosis, antioxidation, and metabolic suppression. Increased availability of NO may be especially important during life-history periods when organisms contend with multiple stresses. We investigated dynamics of the NO metabolites, nitrite (NO2-) and nitrate (NO3-), in the blood plasma, heart, liver, and skeletal muscle of the wood frog (Rana sylvatica), an amphibian that endures chronic cold, freezing, hypoxia, dehydration, and extended aphagia during hibernation. We found elevated concentrations of NO2- and/or NO3- in the plasma (up to 4.1-fold), heart (3.1-fold), and liver (up to 4.1-fold) of frogs subjected to experimental hypoxia (24 h, 4 °C), and in the liver (up to 3.8-fold) of experimentally frozen frogs (48 h, - 2.5 °C), suggesting that increased NO availability aids in survival of these stresses. During a 38-week period of simulated hibernation, NO2- and/or NO3- increased in the plasma (up to 10.4-fold), heart (up to 3.3-fold), and liver (5.0-fold) during an initial 5-week winter-acclimatization regimen and generally remained elevated thereafter. In hibernation, plasma NO2- was higher in frogs indigenous to Interior Alaska than in conspecifics from a temperate locale (southern Ohio), suggesting that NO availability is matched to the severity of environmental conditions prevailing in winter. The comparatively high NO availability in R. sylvatica, a stress-tolerant species, together with published values for other species, suggest that the NO protection system is of general importance in the stress adaptation of vertebrates.


Does Subclinical Hypothyroidism Affect Dynamic Thiol / Disulfide Homeostasis and İschemia-modified Albumin Levels in Children?
To determine the effects of subclinical hypothyroidism on oxidative stress in children. A cross-sectional study. Department of Paediatrics, Paediatric Endocrinology, and General Outpatient Clinics, Kirikkale University, School of Medicine, from May 2017 to October 2018. This study included 92 subjects aged between 2 and 18 years. The subjects were divided into two groups. Forty-seven children with subclinical hypothyroidism and 45 healthy controls were evaluated. In order to evaluate oxidative damage, native thiol, total thiol, disulfides, their ratios, and ischemia-modified albumin (IMA) levels were compared between the two groups. The relationship between TSH and IMA levels was assessed. Age and gender were not significantly different in the two groups. Native thiol, total thiol, disulfides and their ratios were similar in the two groups. Ischemia-modified albumin levels were significantly higher in the patient group than the controls (p<0.001). There was no correlation between TSH and IMA levels in the patient group (r=0.069 p=0.645). Subclinical hypothyroidism may be related to the impairment of IMA, and have a neutral effect on thiol/disulfide balance. Further research is needed to explain the effects of oxidative stress in subclinical hypothyroidism. Key Words: Subclinical hypothyroidism, Childhood, Oxidative stress.


Cardiac aorta-derived extracellular matrix scaffold enhances critical mediators of angiogenesis in isoproterenol-induced myocardial infarction mice.
An incapability to improve lost cardiac muscle caused by acute ischemic injury remains the most important deficiency of current treatments to prevent heart failure. We investigated whether cardiomyocytes culturing on cardiac aorta-derived extracellular matrix scaffold has advantageous effects on cardiomyocytes survival and angiogenesis biomarkers' expression. Ten male NMRI mice were randomly divided into two groups: (1) control (healthy mice) and (2) myocardial infarction (MI)-induced model group (Isoproterenol/subcutaneously injection/single dose of 85 mg/kg). Two days after isoproterenol injection, all animals were sacrificed to isolate cardiomyocytes from myocardium tissues. The fresh thoracic aorta was obtained from male NMRI mice and decellularized using 4% sodium deoxycholate and 2000 kU DNase-I treatments. Control and MI-derived cardiomyocytes were seeded on decellularized cardiac aorta (DCA) considered three-dimensional (3D) cultures. To compare, the isolated cardiomyocytes from control and MI groups were also cultured as a two-dimensional (2D) culture system for 14 days. The cell viability was examined by MTT assay. The expression levels of Hif-1α and VEGF genes and VEGFR1 protein were tested by real-time PCR and western blotting, respectively. Moreover, the amount of VEGF protein was evaluated in the conditional media of the 2D and 3D systems. The oxidative stress was assessed via MDA assay. Hif-1α and VEGF genes were downregulated in MI groups compared to controls. However, the resulting data showed that decellularized cardiac aorta matrices positively affect the expression of Hif-1α and VEGF genes. The expression level of VEGFR1 protein was significantly (p ≤ 0.01) upregulated in both MI and healthy cell groups cultured on decellularized cardiac aorta matrices as a 3D system compared to the MI cell group cultured in the 2D systems. Furthermore, MDA concentration significantly decreased in 3D-cultured cells (MI and healthy cell groups) rather than the 2D-cultured MI group (p ≤ 0.015). The findings suggest that cardiac aorta-derived extracellular scaffold by preserving VEGF, improving the cell viability, and stimulating angiogenesis via upregulating Hif-1α, VEGF, and VEGFR1 in cardiomyocytes could be considered as a potential approach along with another therapeutic method to reduce the complications of myocardial infarction and control the progressive pathological conditions related to MI.


Subcellular distribution and properties of progesterone (delta4-steroid) 5alpha-reductase in rat medial basal hypothalamus.
The subcellular distribution and properties of rat hypothalamic progesterone 5 alpha-reductase, which accelerates the conversion of progesterone to 5 alpha-pregnane-3,20-dione, have been investigated by utilizing 3H-labeled substrate and a reverse isotopic dilution assay system. The enxymic activity was associated primarily with a cell debris-membranes fraction deribed from the 100 x g pellet. This fraction contained mainly membrane-like particulates and was free of nuclei. Little or no activity was associated with the purified nuclei. The hypothalamic 5 alpha-reductase was stimulated by NADPH but not by NADH. The reaction proceeded optimally over a pH range of 6.0 to 7.2 and at a temperaturhe substrate specificity of the enzyme for other delta 4-3-ketosteroids and the ability of these steroids to inhibit the 5 alpha reduction of [1,2-3H]progesterone as well as the effect of 17 beta-estradiol were also studied. 20 alpha-hydroxypregn-4-en-3-one was more reactive that progesterone, while testosterone was the least reactive. The estimated Km for 20 alpha-hydroxypregn-4-en-3-one was 8.6 +/- 1.9 x 10(-7) M, and for testosterone, 1.6 +/- 1.4 x 10(-5) M. The inhibition studies indicate that 20 alpha-hydroxypregn-4-en-3-one and 17 beta-estradiol are competitive and noncompetitive inhibitors, respectively, of the 5 alpha reduction of progesterone with Ki of 6.0 +/- 3.0 x 10(-8) M for 20 alpha-hydroxypregn-4-en-3-one and Kii (intercept inhibition constant) of 2.6 +/- 0.7 x 10(-5) M and Kis (slope inhibition constant) of 3.6 +/- 0.6 x 10(-5) M for 17 beta-estradiol. Testosterone is a poor competitive inhibitor of the reaction.


Retinoic Acid Has Neuroprotective effects by Modulating Thioredoxin in Ischemic Brain Damage and Glutamate-exposed Neurons.
Ischemic stroke is a neurological disorder that causes pathological changes by increasing oxidative stress. Retinoic acid is one of the metabolites of vitamin A. It regulates oxidative stress and exerts neuroprotective effects. Thioredoxin is a small redox protein with antioxidant activity. The aim of this study was to investigate whether retinoic acid modulates the expression of thioredoxin in ischemic brain injury. Cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) surgery and retinoic acid (5 mg/kg) or vehicle was administered to adult male rats for four days prior to surgery. MCAO induced neurological deficits and increased oxidative stress and retinoic acid attenuated these changes. Retinoic acid ameliorated the MCAO-induced decrease in thioredoxin expression. MCAO decreases the interaction between thioredoxin and apoptosis signal-regulating kinase 1 (ASK1), and retinoic acid treatment alleviates this decrease. Glutamate (5 mM) exposure induced cell death and decreased thioredoxin expression in cultured neurons. Retinoic acid treatment attenuated these changes in a dose-dependent manner. Retinoic acid prevented the decrease of bcl-2 expression and the increase of bax expression caused by glutamate exposure. Moreover, retinoic acid attenuated the increases in caspase-3, cleaved caspase-3, and cytochrome c in glutamate-exposed neurons. However, the mitigation effects of retinoic acid were lower in thioredoxin siRNA-transfected neurons than in non-transfected neurons. These results demonstrate that retinoic acid regulates oxidative stress and thioredoxin expression, maintains the interaction between thioredoxin and ASK1, and modulates apoptosis-associated proteins. Taken together, these results suggest that retinoic acid has neuroprotective effects by regulating thioredoxin expression and modulating apoptotic pathway.


MdHIR proteins repress anthocyanin accumulation by interacting with the MdJAZ2 protein to inhibit its degradation in apples.
In higher plants, jasmonate ZIM-domain (JAZ) proteins negatively regulate the biosynthesis of anthocyanins by interacting with bHLH transcription factors. However, it is largely unknown if and how other regulators are involved in this process. In this study, the apple MdJAZ2 protein was characterized in regards to its function in the negative regulation of anthocyanin accumulation and peel coloration. MdJAZ2 was used as a bait to screen a cDNA library using the yeast two-hybrid method. The hypersensitive induced reaction (HIR) proteins, MdHIR2 and MdHIR4, were obtained from this yeast two-hybrid. The ZIM domain of MdJAZ2 and the PHB domain of the MdHIR proteins are necessary for their interactions. The interactions were further verified using an in vitro pull-down assay. Subsequently, immunoblotting assays demonstrated that MdHIR4 enhanced the stability of the MdJAZ2-GUS protein. Finally, a viral vector-based transformation method showed that MdHIR4 inhibited anthocyanin accumulation and fruit coloration in apple by modulating the expression of genes associated with anthocyanin biosynthesis.


A phase III randomized, controlled study to assess and compare the immunogenicity and tolerability of single and multi-dose vials of DTwP-Hib, a fully liquid quadravalent vaccine and their comparison with TETRAct-Hib vaccine in Indian infants aged 6-14 weeks.
Both WHO and IAP encourage using combination vaccines, wherever feasible. The phase III trial reported here was conducted to assess and compare the immunogenicity, tolerability and safety of two quadravalent vaccines, Quadrovax(®) (new vaccine), and TETRAct-Hib(®) (available in the market) in a multicentre study, in India. In all, 361 infants aged 6-8 weeks were enrolled, out of which 339 completed the study. The vaccination was done at 6-10-14 weeks following EPI/WHO recommended immunization schedule. Blood samples were collected prior to the administration of first dose and one month after the third dose. Postvaccination, geometric mean titres for each component did not differ significantly between the single dose vial and multi dose vial subgroups and among the two study groups. Adverse events observed were within the range quoted in literature. Quadrovax(®) vaccine manufactured by SIIL was found to be safe, immunogenic and non-inferior to the comparator vaccine. The quadravalent vaccine is best recommended in the second year of life when children receive their booster dose at 15-18 months. It can be given to infants during primary immunization series at 6, 10 and 14 weeks of age when Hepatitis B vaccine is given in a separate arm or to infants at 10 weeks who receive the Hepatitis B vaccine separately following the 0, 6 and 14 weeks or 0, 1 and 6 months schedule.


Effect of smoking on oxygen delivery and outcome in patients treated with radiotherapy for head and neck squamous cell carcinoma--a prospective study.
Head and neck cancer patients with high hemoglobin respond better to irradiation compared to patients with low hemoglobin possibly due to hypoxia induced radioresistance. The hemoglobin level is, however, a crude indicator of the amount of oxygen available to the tissue and may be influenced by a number of factors, smoking being of potential importance. The aim of the present study was to examine the effect of smoking on available oxygen to tumors and the effect on outcome in head and neck cancer patients treated with radiotherapy in a prospective study. A total of 232 consecutive patients with squamous cell carcinoma of the larynx, pharynx and oral cavity completed questionnaires on smoking habits prior to treatment. Venous blood samples were collected before and/or during treatment to determine the hemoglobin and carboxyhemoglobin level. Patients were treated with primary curative radiotherapy 62-68 Gy, 2 Gy/fx, 5 fx/week. All but 12 patients had a history of smoking, 35 were long term quitters, 23 recent quitters, 54 moderate smokers and 108 heavy smokers (>1 pack/day). There was no relationship between total hemoglobin and carboxyhemoglobin, but effective hemoglobin and carboxyhemoglobin were linearly correlated. The amount of carboxyhemoglobin increased with increasing smoking status. Actuarial 5-year univariate analysis showed that heavy smokers had a significantly reduced probability of loco-regional control (44% vs. 65%, p = 0.001), disease-specific (56% vs. 77%, p = 0.003) and overall survival (39% vs. 66%, p = 0.0004) compared to non-smoking patients. Multivariate analyses showed that patients characterized as non-smokers, with low T and N classifications and high hemoglobin level had the best outcome measurements. A rise in carboxyhemoglobin significantly decreased the probability of loco-regional control and each additional pack year increased the risk of death. Smokers and former smokers develop secondary cancers. The study showed a significant negative impact of smoking during radiotherapy for head and neck cancer and the risk of death was increased with each additional pack year of smoking. The effect on loco-regional control could be explained by a rise in carboxyhemoglobin level in smokers, e.g. a reduced oxygen supply to tumors. The data strongly advocate that smoking should be avoided in order to improve the therapeutic efficacy of radiotherapy and development of other smoking-related diseases and/or secondary cancers.


Curbing dioxin emissions from municipal solid waste incineration: China's action and global share.
China generates the world's second-largest amount of municipal solid waste (MSW) and incinerates the largest quantity of MSW. However, data on the latest dioxin emissions from MSW incineration (MSWI) and the related global share were lacking. In the context of MSW classification, distinguishing the long-term MSW generation and incineration quantity, and dioxin emissions was necessary for macro-control and policy-making by the Chinese Government. By considering population size and GDP per capita, China's MSW generation toward 2050 was projected based on Monte Carlo simulation. Moreover, dioxin emission factors were also assumed based on the diffusion rate of four grades of air pollution control devices (APCDs). Finally, we show that the quantity of China's MSW generation in 2050 will be 363.50 million tonnes (Mt) with 341.06-382.45 Mt of 75% certainty. China's dioxin emissions from MSWI were approximately 15.46 g I-TEQ in 2019, which accounted for 26.1% of total emissions from global MSWI. We discuss dioxin emission reduction scenarios depending on MSW diversion and APCD upgrades. China's dioxin emissions will be 70.38 g I-TEQ for the business-as-usual scenario, and the dioxin emissions will be 9.29 g I-TEQ (within the range of 8.88-9.64 g I-TEQ) for the optimal scenario in 2050. Moreover, in 2050, the APCD diffusion rate will account for 98.8% of the sensitivity of dioxin emissions from China's MSWI. According to the assumed scenarios, there is a dioxin emission reduction potential of 18.6% and 86.8% in 2050 by MSW diversion alone and maximum APCD upgrades combined with food waste diversion, respectively.


Theoretical approach to local infusion of antibiotics for infected pancreatic necrosis.
Infected pancreatic necrosis is a major complications of acute pancreatitis. If drainage is required, local administration of antibiotics through transmural nasocystic or percutaneous catheter may allow increasing local antibiotic concentrations. Drug diffusion becomes the main factor influencing local drug tissue penetration. The present study aims at providing the rationale for the design of new research protocols evaluating the efficacy of local antibiotics for infected pancreatic necrosis. A review of microbiological data was performed for the most common organisms causing the infection, antibiotics spectrum and minimum inhibitory concentrations (MIC). A search of the physico-chemical properties of antibiotics was performed to calculate the diffusion coefficients. An estimation of the antibiotic concentrations in pancreatic tissue was obtained using a mathematical model. Efficacy factors (EF) were calculated and the stability of the antibiotic solutions were evaluated to optimize the dosing regimen. Piperacillin, vancomycin and metronidazole achieve high concentrations in the surrounding tissue very fast. Imipenem, ceftriaxone, ciprofloxacin, gentamicin, linezolid and cloxacillin achieve intermediate concentration values. Tigecycline, showed the lowest concentration values (<2 mg/L). Calculated EF is highest for piperacillin and imipenem short after administration and near to surface diffusion area (0.5 cm), but EF of imipenem is higher at deeper areas and longer time after administration. Considering obtained results, some solutions are proposed using saline as diluent and 25 °C of temperature during administration. Imipenem has the best theoretical results in empiric local treatment. Linezolid and tigecycline solutions are not recommended.


Effect of hypophysectomy on cAMP changes in rat adrenal medulla evoked by catecholamines and carbamylcholine.
It was the aim of this study to investigate the mechanisms responsible for changes in 3',5'-cyclic adenosine monophosphate (cAMP) in the rat adrenal medulla occuring after administration of carbamylcholine, histamine, ACTH and various phenylethylamines. Carbamylcholine, ACTH, histamine, noradrenaline and dopamine produced marked (500-900%) increases in adrenal cAMP which were very similar in both adrenal cortex and medulla both with respect to time-course and relative extent. Interestingly isoprenaline and adrenaline did not influence cAMP levels even at excessively high doses. In all cases studied transsection of the splanchnic fibers supplying the adrenals reduced the increase in medullary cAMP by not more than 25--30%, suggesting that cAMP levels in the adrenal medulla are predominantly regulated by non-neuronal mechanisms. This assumption was strongly supported by the observation that hypophysectomy completely abolished the 500--600% increase in cAMP produced by 50 mumol/kg of dopamine and reduced the 700% increase resulting from 4.4 mumol/kg of carbamylcholine to 70%. In spite of the marked increase in cAMP produced by single and repeated doses of dopamine in the adrenal medulla there was no subsequent induction of tyrosine hydroxylase (TH). Moreover carbamylcholine (8.2 mumol/kg) evoked TH induction only in innervated adrenals whereas after denervation, in spite of the large (+ 500%) and prolonged (more than 90 min) increase in cAMP, no TH induction could be observed. It is concluded that adrenal medullary cAMP is predominantly regulated by the pituitary gland via the adrenal cortex and only to a much smaller extent--if at all--by direct cholinergic mechanisms, which are responsible for the initiation of TH induction.


Residents' perceptions of the ideal clinical teacher--a qualitative study.
The object of this study was to establish what residents in 1994 and 2003 characterised as an ideal clinical teacher and whether differences existed between residents' views in 1994 and 2003. postgraduate medical education in the Netherlands. 207 obstetric-gynaecologic residents. open-ended questionnaire. qualitative data analysis with two coding dictionaries based on current literature. Differences between 1994 and 2003 were estimated using the Chi-square test. Residents preferred the 'person' role both in 1994 (42%) and in 2003 (48%). The 'physician' role was significantly more important in 1994 than in 2003; the 'supervisor' role was significantly more important in 2003 than in 1994 (p<0.05). Seventy percent of the comments related to 'direct interaction' (i.e., between residents and clinical teachers), 30% to 'indirect interaction' (i.e., clinical teachers' behaviour affecting residents indirectly). The data showed that almost half of residents' comments described 'person' role characteristics. There was a significant shift in the role ranked second, from the physician role in 1994 to the supervisor role in 2003. The findings highlighted that teachers, in order to be perceived as ideal, should adapt their behaviour to residents' learning needs.


Clinical practice of health professionals working in early detection for infants with or at risk of cerebral palsy across New Zealand.
A diagnosis of cerebral palsy (CP) can, and should, be made as early as possible. This work describes current clinical practice around the awareness and use of diagnostic tools for the detection of CP in New Zealand (NZ). A purpose-developed survey distributed electronically to NZ clinicians working with young children with or at risk of CP. A total of 159 clinicians (including paediatricians, physiotherapists and occupational therapists) participated in this cross-sectional study. Ninety-six percent were aware that a diagnosis of CP can be made by 12 months of age, with high levels of awareness of the use of magnetic resonance imaging (94%), Prechtl's qualitative assessment of general movements (GMs) (70%) and Hammersmith Infant Neurological Examination (HINE) (77%). Only 40% were aware of the HINE optimality scoring. Fifty-four clinicians provided a diagnosis of CP as part of their role: 48% never used the GMs or HINE to assess children <1 year, and 57% never used the HINE for children between 1 and 2 years. Clinicians not providing a diagnosis within their professional role (n = 104) also indicated infrequent use of assessment tools with 74% and 54% never using the GM's or HINE (respectively) in their assessment of children at risk of CP. Barriers to use included lack of time and funding, lack of clear pathways and management support. Despite high awareness, current use of international best practice tools in NZ clinical practice appears low. Multiple barriers are reported to the use of these tools, which need to be addressed to improve the timeliness of diagnosis.


Weight loss, ion release and initial mechanical properties of a binary calcium phosphate glass fibre/PCL composite.
Composites comprising a biodegradable polymeric matrix and a bioactive filler show considerable promise in the field of regenerative medicine, and could potentially serve as degradable bone fracture fixation devices, depending on the properties obtained. Therefore, glass fibres from a binary calcium phosphate (50P(2)O(5)+50CaO) glass were used to reinforce polycaprolactone, at two different volume fractions (V(f)). As-drawn, non-treated and heat-treated fibres were assessed. Weight loss, ion release and the initial mechanical properties of the fibres and composites produced have been investigated. Single fibre tensile testing revealed a fibre strength of 474MPa and a tensile modulus of 44GPa. Weibull analysis suggested a scale value of 524. The composites yielded flexural strength and modulus of up to 30MPa and 2.5GPa, respectively. These values are comparable with human trabecular bone. An 8% mass loss was seen for the lower V(f) composite, whereas for the two higher V(f) composites an approximate 20% mass loss was observed over the course of the 5week study. A plateau in the degradation profile at 350h indicated that fibre dissolution was complete at this interval. This assertion was further supported via ion release studies. The leaching of fibres from the composite created a porous structure, including continuous channels within the polymer matrix. This offers further scope for tailoring scaffold development, as cells from the surrounding tissue may be induced to migrate into the resulting porous matrix.


Inhibition of TRAF6 improves hyperlipidemic acute pancreatitis by alleviating pyroptosis in vitro and in vivo rat models.
Hypertriglyceridemia (HTG) is one of the common causes of acute pancreatitis (AP). Hyperlipidemic acute pancreatitis (HTG-AP) is associated with higher mortality owing to its tendency for greater severity and rapid progression. The purpose of this study was to explore the mechanism of involvement of tumor necrosis factor receptor-related factor 6 (TRAF6) in pyroptosis during HTG-AP. The HTG environment was simulated with palmitic acid treatment in vitro and a high-fat diet in vivo. Cerulein was used to establish the HTG-AP model, followed by genetic and pharmacological inhibition of TRAF6. Pyroptosis activation, inflammatory reaction, and the interaction between TRAF6 and pyroptosis in HTG-AP were assessed. HTG was found to aggravate the development of pancreatitis, accompanied by increased pyroptosis and enhanced inflammatory response in HTG-AP models. Mechanistically, TRAF6 downregulation decreased the activation of pyroptosis in cerulein-induced HTG-AP. Collectively, inhibition of TRAF6 improved HTG-AP and the associated inflammation by alleviating pyroptosis.


Isolation, growth, ultrastructure, and metal tolerance of the green alga, Chlamydomonas acidophila (Chlorophyta).
An acidophilic volvocine flagellate, Chlamydomonas acidophila (Volvocales) that was isolated from an acid lake, Katanuma, in Miyagi prefecture, Japan was studied for growth, ultrastructural characterization, and metal tolerance. Chlamydomonas acidophila is obligately photoautotrophic, and did not grow in the cultures containing acetate or citrate even in the light. The optimum pH for growth was 3.5-4.5. To characterize metal tolerance, the toxic effects of Cd, Co, Cu, and Zn on this alga were also studied. Effective metal concentrations, which limited the growth by 50%, EC50 were measured, after 72 h of static exposure. EC50s were 14.4 microM Cd2+, 81.3 microM Co2+, 141 microM Cu2+, and 1.16 mM Zn2+ for 72 h of exposure. Thus, this alga had stronger tolerance to these metals than other species in the genus Chlamydomonas.


The MCSS-26: revision of the Manchester Clinical Supervision Scale using the Rasch Measurement Model.
Previously published accounts of the evaluation of the effects of clinical supervision, a structured system to support health service staff, have been mainly contained to small scale qualitative studies. Over the past decade, the 36-item Manchester Clinical Supervision Scale (MCSS) has transformed the evaluation landscape and has been used as a quantitative outcome measure in upward of 90 licensed studies in 12 countries worldwide. The factor structure has been replicated by other researchers and the psychometric properties have been found robust. However, it had not been previously tested empirically using newly available and sophisticated statistical analyses. This study tested the original factor structure and response format of the MCSS for goodness of fit to the Rasch model, using Rasch Unidimensional Measurement Model (RUMM) 2030 software, and investigated the validity of the questionnaire for both nursing and allied health (AH) staff. A series of Rasch analyses were conducted on the seven subscales of the MCSS. The default procedure for RUMM software uses the partial credit model, which allows items to have varying numbers of response categories and does not assume the distance between response thresholds is uniform. Detailed Rasch analyses indicated that the 36-item version of the MCSS could be reduced to 26 items and result in improved fit statistics for six subscales rather than seven. This study reconfirmed the established psychometric properties of the MCSS, now renamed the MCSS-26.


Newly-developed measuring device for the quantitative assessment of thermal and pain thresholds of peripheral nerves.
Measuring devices for the quantitative assessment of thermal and pain thresholds are either simple and only suited for inexact tests or accurate and objective but expensive. The aim was there for to develop a cost effective device to enable a short and practical test of neurosensibility under clinical conditions. The result of this development is a new thermosensibility-measuring device (TSM) consisting of the measuring unit itself and a thermal probe. The data are registered and analysed through direct data transfer to a connected PC. Investigations carried out with this device revealed the construction to be efficient and easy to handle under clinical conditions. The TSM provides the examiner with the opportunity to monitor the neurosensory function of peripheral nerves in a reproducible way.


Indications for surgical treatment of cerebrovascular insufficiency.
197 cases of patients with CVI, subjected to surgery by carotid endarterectomy, are examined. The results were nearly always good and hence the technique is recommended. Particular interest is attached to the risk connected with carotid clamping, through intraoperative study of CBF with 133Xe, such a risk is practically removed.


Enhanced clearance of a multiple antibiotic resistant Staphylococcus aureus in rats treated with PGG-glucan is associated with increased leukocyte counts and increased neutrophil oxidative burst activity.
PGG-Glucan [Betafectin], a highly purified soluble beta-(1-6)-branched beta-(1 3)-linked glucan isolated from Saccharomyces cerevisiae, has broad in vitro and in vivo anti-infective activities unrelated to cytokine induction. Here we present in vivo results on the anti-infective activity of PGG-Glucan against a multiple antibiotic resistant Staphylococcus aureus. PGG-Glucan (0.25-4 mg/kg) was administered intramuscularly to male Wistar rats 48 h, 24 h, and 4 h before and 4 h after intraperitoneal implantation of a gelatin capsule containing 10(8)S. aureus colony forming units (CFU). Blood samples were collected at various times after challenge to determine CFU levels, leukocyte counts and neutrophil oxidative burst activity; serum TNF-alpha, and IL-1beta levels were also evaluated. The 0.25 mg/kg PGG-Glucan dose had no effect on reducing blood CFU levels; however, PGG-Glucan doses of 0.5 mg/kg, 1 mg/kg, 2 mg/kg or 4 mg/kg significantly reduced blood CFU levels by 48 h after challenge. Reduced CFU levels correlated with significantly elevated absolute monocyte counts, absolute neutrophil counts, and neutrophil oxidative burst activity in the absence of any effect on TNF-alpha or on IL-1beta levels. In additional studies, effects on mortality and blood CFU levels were evaluated in rats treated with ampicillin (an antibiotic to which the S. aureus was resistant), PGG-Glucan, or both agents. Mortality and blood CFU levels were reduced most in combination-treated rats compared to saline control rats or rats treated with either ampicillin alone or PGG-Glucan alone. We conclude that in vivo (1) PGG-Glucan can enhance clearance of an antibiotic resistant S. aureus, (2) that this clearance is accompanied by an increase in monocytes and neutrophils as well as a potentiation of neutrophil oxidative microbiocidal activity without alteration of the proinflammatory cytokine response, and (3) PGG-Glucan can enhance the effectiveness of traditional antibiotic treatment.


Chronic noise stress and dexamethasone administration on blood pressure elevation in the rat.
In order to evaluate the role of ACTH and corticosterone in the elevated blood pressure produced by chronic noise, we have studied the effect of dexamethasone administration on blood pressure. Adrenal response was measured by alterations in plasma corticosterone levels in two experimental groups (Dexamethasone treated rats, Dexamethasone-noise stressed rats) and compared with a Control and a Noise-stressed group. Chronic noise stress with a frequency of 2640 Hz, power of 30 w and duration of 15 min daily was used for 30 consecutive days. Dexamethasone was administered by subcutaneous injection of 100 micrograms/100 g b.wt daily. Blood pressure was measured by an indirect tail cuff method and corticosterone levels by specific RIA. Dexamethasone administration decreases corticosterone levels but increases blood pressure. Dexamethasone-treated noise stressed rats show higher residual corticosterone levels and a more marked increase on blood pressure than rats treated with dexamethasone alone. Thus noise-stress and dexamethasone administration have opposing effects on corticosterone release and a synergistic effect on blood pressure elevation.