Effect of primer-cement systems with different functional phosphate monomers on the adhesion of zirconia to dentin.
The objective of this study was to evaluate the effect of primer-cement systems with different functional phosphate monomers on the adhesion of zirconia to dentin with and without aging protocols. Bovine teeth (N = 180) were embedded in acrylic resin after sectioning their roots with with their coronal parts exposed. The buccal surface of each tooth was polished with silicon carbide papers (#200, 400, 600) until dentin exposure. Sintered zirconia cylinders (N = 180) (Ø: 3.4 mm; height: 4 mm) (Vita In-Ceram 2000) were prepared and distributed into 18 groups (n = 10 per group) considering the following factors: "Cementation System" (Panavia F - PAN; RelyX Ultimate - ULT, Multilink N - MULT) and "aging" (water storage in distilled water at 37 °C for 24 h (control, C); 30 days (30D); 6 months (6 M) and thermocycling for 5000 (5TC), 10,000 (10TC) and 20,000 (20TC) thermal cycles (5-55 °C; dwell time: 30 s)". Zirconia and dentin cementation surfaces were conditioned according to the recommendations of the manufacturers of each resin cement. The cylinders were adhesively cemented to the dentin surfaces and the specimens were submitted to the aging protocols. After aging, the specimens were subjected to shear bond strength test (SBS) (1 mm/min) in a Universal Testing Machine and failure types were analyzed. The data (MPa) were statistically using Kruskal-Wallis followed by the Dunn test (α = 5%). The degree of conversion (DC) rates of the cementing systems were also measured. While without aging (24 h) no significant difference was found between the cement systems (p > 0.05), after 30D (4.3-5.4), the highest decrease in all groups were observed after 5TC (1.5-2.3) (p < 0.05). Overall, MULT and ULT presented significantly higher results than that of PAN (p < 0.05). Pre-test failures during TC were more frequent in the PAN group. Complete adhesive failures at the cement/dentin interface were more frequent for MULT (30-80%) and PAN (10-70%) and for ULT (20-90%) at the cement/ceramic interface. DC of the tested cements did not show significant difference. Adhesion performance of the primer-cement systems with different functional phosphate monomers on zirconia-dentin complex varied as a function of aging strategies with MULT and ULT delivering higher bond strength values. When failure types considered, none of the cement systems performed well on both ceramic and dentin.


Spontaneous bacterial peritonitis caused by Campylobacter Coli in cirrhotic patient: A rare case report (CARE-compliant).
Spontaneous bacterial peritonitis (SBP) is a fatal infection in patients. It often happens in patients with cirrhosis, cancer or diabetes, and is caused mostly by Enterobacteriaceae. Here we report a rare case of SBP caused by Campylobacter Coli (C coli) infection, which was identified promptly by the matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and received adequate therapy sooner after. In the present study, we reported a 46-year-old male with alcoholic liver cirrhosis (Child-Pugh class C) and type 2 diabetes mellitus presented with a 1-day history of fever and abdominal pain. Based on the clinical examinations, the patient was diagnosed with SBP and the pathogen was quickly identified as C coli by the matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), a rare causative pathogen of SBP. The patient received a 10-day antibiotic treatment with Ciprofloxacin 400 mg every 12 hours, and recovered successfully. The patient had a successful treatment outcome. The study demonstrated a new possible infectious cause of SBP by C Coli, which was rarely seen in liver cirrhosis but mostly found in immunocompromised patients. Thus, it might raise an idea of microorganism screening of broader types that might also induce SBP for immunocompromised patients.


Reoperation for stenotic saphenous vein bypass grafts without cardiopulmonary bypass or shunt.
From October 1982 through February 1985, six patients have undergone seven reoperative revascularization procedures for stenosis of coronary artery vein bypass grafts without the use of cardiopulmonary bypass or intraoperative shunt. All patients had prior coronary artery saphenous vein bypasses, ranging from 2 months to 5 years preoperatively. All patients survived reoperation and there was no morbidity. Patients have been followed-up to 26 months. One patient underwent postoperative cardiac catheterization at 1 year, which revealed a patent interposition graft, and one patient had a second operation for restenosis of a vein graft. To decrease the risk of morbidity and mortality in reoperative procedures for saphenous vein graft stenosis, a technique is described that avoids the use of cardiopulmonary bypass or intraoperative shunt. This technique has been used successfully for seven reoperative revascularization procedures and is recommended for selected cases.


Course of migraine during pregnancy and postpartum: a prospective study.
The aim of this study was to investigate prospectively the course of migraine during pregnancy and postpartum. Of all the pregnant women consecutively attending an obstetrics and gynaecology department for a routine first-trimester antenatal check-up, 49 migraine sufferers--two were affected by migraine with aura (MA) and 47 by migraine without aura (MO)--who had experienced at least one attack during the 3 months preceding pregnancy were identified, enrolled in the study and given a headache diary. Subsequent examinations were performed at the end of the second and third trimesters and 1 month after delivery. Migraine was seen to improve in 46.8% of the 47 MO sufferers during the first trimester, in 83.0% during the second and in 87.2% during the third, while complete remission was attained by 10.6%, 53.2%, and 78.7% of the women, respectively. Migraine recurred during the first week after childbirth in 34.0% of the women and during the first month in 55.3%. Certain risk factors for lack of improvement of migraine during pregnancy were identified: the presence of menstrually related migraine before pregnancy was associated with a lack of headache improvement in the first and third trimesters, while second-trimester hyperemesis, and a pathological pregnancy course were associated with a lack of headache improvement in the second trimester. Breast feeding seemed to protect from migraine recurrence during postpartum.


Specificity and stability of the Acromyrmex-Pseudonocardia symbiosis.
The stability of mutualistic interactions is likely to be affected by the genetic diversity of symbionts that compete for the same functional niche. Fungus-growing (attine) ants have multiple complex symbioses and thus provide ample opportunities to address questions of symbiont specificity and diversity. Among the partners are Actinobacteria of the genus Pseudonocardia that are maintained on the ant cuticle to produce antibiotics, primarily against a fungal parasite of the mutualistic gardens. The symbiosis has been assumed to be a hallmark of evolutionary stability, but this notion has been challenged by culturing and sequencing data indicating an unpredictably high diversity. We used 454 pyrosequencing of 16S rRNA to estimate the diversity of the cuticular bacterial community of the leaf-cutting ant Acromyrmex echinatior and other fungus-growing ants from Gamboa, Panama. Both field and laboratory samples of the same colonies were collected, the latter after colonies had been kept under laboratory conditions for up to 10 years. We show that bacterial communities are highly colony-specific and stable over time. The majority of colonies (25/26) had a single dominant Pseudonocardia strain, and only two strains were found in the Gamboa population across 17 years, confirming an earlier study. The microbial community on newly hatched ants consisted almost exclusively of a single strain of Pseudonocardia while other Actinobacteria were identified on older, foraging ants in varying but usually much lower abundances. These findings are consistent with recent theory predicting that mixtures of antibiotic-producing bacteria can remain mutualistic when dominated by a single vertically transmitted and resource-demanding strain.


[Features of peptic ulcer with concomitant bronchial asthma in the elderly].
The purpose of the study was to examine particular features of the clinical course of stomach ulcer (SU) and duodenal ulcer (DU) in conjunction with bronchial asthma (BA) in elder patients. We examined 59 patients with BA in conjunction with SU and DU at the age of 69-89 (46 males and 13 females). All patients underwent esophagogastroduodenoscopy (EGDS) and H. pylori examination of tissue samplings from the mucous coat of the stomach and duodenum (urease test). BA was diagnosed based on the clinical picture, examination of the external respiration function, reversibility of bronchial obstruction of not less than 20% after the pharmacological test or anti-inflammatory treatment.


Regulation of lamp2a levels in the lysosomal membrane.
The selective degradation of cytosolic proteins in lysosomes by chaperone-mediated autophagy depends, at least in part, on the levels of a substrate receptor at the lysosomal membrane. We have previously identified this receptor as the lysosome-associated membrane protein type 2a (lamp2a) and showed that levels of lamp2a at the lysosomal membrane directly correlate with the activity of the proteolytic pathway. Here we show that levels of lamp2a at the lysosomal membrane are mainly controlled by changes in its half-life and its distribution between the lysosomal membrane and the matrix. The lysosomal degradation of lamp2a requires the combined action of at least two different proteolytic activities at the lysosomal membrane. Lamp2a is released from the membrane by the action of these proteases, and then the truncated lamp2a is rapidly degraded within the lysosomal matrix. Membrane degradation of lamp2a is a regulated process that is inhibited in the presence of substrates for chaperone-mediated autophagy and under conditions that activate that type of autophagy. Uptake of substrate proteins also results in transport of some intact lamp2a from the lysosomal membrane into the matrix. This fraction of lamp2a can be reinserted back into the lysosomal membrane. The traffic of lamp2a through the lysosomal matrix is not mediated by vesicles, and lamp2a reinsertion requires the lysosomal membrane potential and protein components of the lysosomal membrane. The distribution of lamp2a between the lysosomal membrane and matrix is a dynamic process that contributes to the regulation of lysosomal membrane levels of lamp2a and consequently to the activity of the chaperone-mediated autophagic pathway.


Gender differences in alcohol-related non-consensual sex; cross-sectional analysis of a student population.
Sexual offences are a global public health concern. Recent changes in the law in England and Wales have dramatically altered the legal landscape of sexual offences, but sexual assaults where the victim is voluntarily intoxicated by alcohol continue to have low conviction rates. Worldwide, students are high consumers of alcohol. This research aimed to compare male and female students in relation to their knowledge and attitudes about alcohol and sexual activity and to identify factors associated with being the victim of alcohol-related non-consensual sex. 1,110 students completed an online questionnaire. Drinking levels were measured using the Alcohol Use Disorder Identification Test. Non-consensual sexual experiences were measured using the Sexual Experience Survey. Univariate and multivariate analyses were undertaken using chi square and backwards stepwise logistic regression respectively. A third of respondents had experienced alcohol-related non-consensual sex. Male and female students differed in the importance they gave to cues in deciding if a person wished to have sex with them and their understanding of the law of consent. 82.2% of women who had experienced alcohol-related non-consensual sex were hazardous drinkers compared to 62.9% who drank at lower levels (P < 0.001). Differences existed between men and women, and between those who had and had not experienced alcohol-related non-consensual sex, in relation to assessments of culpability in scenarios depicting alcohol-related intercourse. A third of respondents believed that a significant proportion of rapes were false allegations; significantly more men than women responded in this way. Alcohol-related coerced sexual activity is a significant occurrence among students; attitudinal and knowledge differences between males and females may explain this. Educational messages that focus upon what is deemed acceptable sexual behaviour, the law and rape myths are needed but are set against a backdrop where drunkenness is commonplace.


[A comparative study of the morphofunctional changes in the thyroid of carp and rats as a result of being in an environment with an elevated lead level].
It has been noticed that the morphofunctional organization of the thyroid gland is similar in representatives of two different classes of vertebrates: in fishes and in rats. Exposure of experimental animals to ecological factors with increased lead levels was followed by a phase response (activation, depression) of various structural elements of the thyroid gland. Obvious differences in morphofunctional mechanisms of adaptation to chemical stress factors in fishes and rats were found. A possible relationship is discussed between the environmental pollution with heavy metals and the increased incidence of thyroid gland disease in human population.


[Cytoarchitecture of the principle diencephalic visual centers in the embryo and the chicken Gallus domesticus L. Experimental study].
The effects of the bilateral removal of the optic vesicle on the cytoarchitectonics of the main diencephalic visual centres of Gallus domesticus L. embryos and chicken have been studied from the 10th, day of incubation until the 15th, day after hatching. - Without afferent optic pathways there is no significnat change in cellular components of nucleus lateralis anterior, nucleus geniculatus lateralis ventralis, nucleus griseus tectalis, nucleus rotundus and nucleus pretectalis principalis. When these centres have specific organization, one may observe a spatial distribution of neurons different from that of unoperated chickens. - Whereas the nucleus ectomamillaris disappears early, the nucleus superficialis synencephali is not differentiated. the nucleus geniculatus lateralis dorsalis, reduced in size, shows an alternation of cell constitution even if cytoarchitectonics appears to be similar to that of unoperated chickens. A comparison of these results with those on tectum opticum by authors brings up the question of the exact nature of retino-thalamic connections and of their morphogenetic role. If morphogenetic influence appears to be certain on some structures it seems to be limited on others.


Removal of Cr(VI) by modified diatomite supported NZVI from aqueous solution: evaluating the effects of removal factors by RSM and understanding the effects of pH.
In this study, a novel composite of modified diatomite supported nanoscale zero-valent iron (mD-NZVI) was synthesized and characterized. The effects of four factors (mD-NZVI dose, temperature, contact time and initial pH) on the removal of Cr(VI) by mD-NZVI were studied by experimental work and analyzed by response surface methodology (RSM). A second-order polynomial equation fitted by Box-Behnken design was used as a statistical model and proved to be precise in describing the significance of four factors. The analysis results show that the effects of four factors on the removal efficiency of Cr(VI) were significant (F value is 19.83), initial pH was found to be the key factor. In addition, the effect of initial pH was further studied and the maximum removal efficiency of 89.34% was obtained at pH of 3, the decrease in removal efficiency with the increase in pH is a synergistic effect of Cr(VI) species, surface charge of mD-NZVI and OH- amount at different pH.


Beyond reasonable drought: hotspots reveal a link between the 'Big Dry' and cryptosporidiosis in Australia's Murray Darling Basin.
There is little evidence on how the health impacts of drought vary spatially and temporally. With a focus on waterborne cryptosporidiosis, we identify spatio-temporal hotspots and by using interrupted time series analysis, examine the impact of Australia's Big Dry (2001-2009) in these disease clusters in the Murray Darling Drainage Basin. Analyses revealed a statistically significant hotspot in the north of the Australian Capital Territory (ACT) and a hotspot in the north-eastern end of the basin in Queensland. After controlling for long-term trend and seasonality in cryptosporidiosis, interrupted time series analysis of reported cases in these hotspots indicated a statistically significant link with the Big Dry. In both areas, the end of the Big Dry was associated with a lower risk of reported cryptosporidiosis; in the ACT, the estimated relative risk (RR) was 0.16 (95% confidence interval: 0.07; 0.33), and in Queensland the RR was 0.42 (95% confidence interval: 0.19; 0.42). Although these data do not establish a causal association, this research highlights the potential for drought-related health risks.


Comparison of ICD code-based diagnosis of obesity with measured obesity in children and the implications for health care cost estimates.
Administrative health databases are a valuable research tool to assess health care utilization at the population level. However, their use in obesity research limited due to the lack of data on body weight. A potential workaround is to use the ICD code of obesity to identify obese individuals. The objective of the current study was to investigate the sensitivity and specificity of an ICD code-based diagnosis of obesity from administrative health data relative to the gold standard measured BMI. Linkage of a population-based survey with anthropometric measures in elementary school children in 2003 with longitudinal administrative health data (physician visits and hospital discharges 1992-2006) from the Canadian province of Nova Scotia. Measured obesity was defined based on the CDC cut-offs applied to the measured BMI. An ICD code-based diagnosis obesity was defined as one or more ICD-9 (278) or ICD-10 code (E66-E68) of obesity from a physician visit or a hospital stay. Sensitivity and specificity were calculated and health care cost estimates based on measured obesity and ICD-based obesity were compared. The sensitivity of an ICD code-based obesity diagnosis was 7.4% using ICD codes between 2002 and 2004. Those correctly identified had a higher BMI and had higher health care utilization and costs. An ICD diagnosis of obesity in Canadian administrative health data grossly underestimates the true prevalence of childhood obesity and overestimates the health care cost differential between obese and non-obese children.


Locus of semantic interference in picture naming: Evidence from dual-task performance.
Disagreement exists regarding the functional locus of semantic interference of distractor words in picture naming. This effect is a cornerstone of modern psycholinguistic models of word production, which assume that it arises in lexical response-selection. However, recent evidence from studies of dual-task performance suggests a locus in perceptual or conceptual processing, prior to lexical response-selection. In these studies, participants manually responded to a tone and named a picture while ignoring a written distractor word. The stimulus onset asynchrony (SOA) between tone and picture-word stimulus was manipulated. Semantic interference in naming latencies was present at long tone pre-exposure SOAs, but reduced or absent at short SOAs. Under the prevailing structural or strategic response-selection bottleneck and central capacity sharing models of dual-task performance, the underadditivity of the effects of SOA and stimulus type suggests that semantic interference emerges before lexical response-selection. However, in more recent studies, additive effects of SOA and stimulus type were obtained. Here, we examined the discrepancy in results between these studies in 6 experiments in which we systematically manipulated various dimensions on which these earlier studies differed, including tasks, materials, stimulus types, and SOAs. In all our experiments, additive effects of SOA and stimulus type on naming latencies were obtained. These results strongly suggest that the semantic interference effect arises after perceptual and conceptual processing, during lexical response-selection or later. We discuss several theoretical alternatives with respect to their potential to account for the discrepancy between the present results and other studies showing underadditivity.


Effects of PTH, calcitonin, and cAMP on calcium transport in rabbit distal nephron segments.
Distal nephron segments are heterogenous with respect to adenylate cyclase responses to stimulation with parathyroid hormone (PTH) or calcitonin (CT). We examined effects of these hormones and of 8-(p-chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (CPTcAMP) on net Ca absorption (Jnet Ca2+, pmol.min-1.mm-1) in rabbit distal nephron segments by in vitro microperfusion technique. We studied three segments, including distal convoluted tubule (DCT), connecting tubule (CNT), and cortical collecting duct (CCD). PTH (1 nM) in bath significantly increased Jnet Ca2+ from 2.28 +/- 0.35 to 9.44 +/- 1.13 in CNT, but did not affect Jnet Ca2+ in DCT or CCD. CT (1 nM) in bath significantly increased Jnet Ca2+ from 1.58 +/- 0.29 to 4.45 +/- 1.01 in DCT, whereas it did not affect Jnet Ca2+ either in CNT or in CCD. CPTcAMP (30 microM) in bath significantly increased Jnet Ca2+ from 2.29 +/- 0.42 to 3.97 +/- 0.43 in DCT and from 2.43 +/- 0.18 to 5.83 +/- 0.37 in CNT, but it did not affect Jnet Ca2+ in CCD. When Na+ was removed from bathing fluid or when 0.1 mM ouabain was added to bath, Jnet Ca2+ in both DCT and CNT significantly decreased. Furthermore, stimulatory effects of PTH and CT on Ca2+ absorption in the respective segments were abolished under these conditions. These results suggest that PTH and CT increase Ca2+ absorption in CNT and DCT, respectively, through cAMP-mediated mechanisms. Presence of a basolateral Na(+)-Ca2+ exchange process seems to be a prerequisite for effects of these hormones. However, exact intracellular mechanisms remain uncertain.


Military sexual trauma in men: a review of reported rates.
Military sexual trauma (MST) has historically been associated with female service members, but it is also experienced by male service members. This article reviews reported prevalence and incidence rates of men's MST in 29 studies. Sources for these studies included the Department of Defense, the U.S. military service academies, and the Department of Veterans Affairs. There is significant variability in reported rates of men's MST. Averaging across studies covering the past 30 years, we found that MST is reported by approximately 0.09% of male service members each year, with a range of 0.02% to 6%. MST is reported by 1.1% of male service members over the course of their military careers, with a range of 0.03% to 12.4%. Determining prevalence and incidence rates for both men's and women's MST is fraught with limitations, including (a) cross-study variations in sample, method, definitions, and assessment and (b) barriers to reporting MST. Each of these limitations is reviewed with an eye toward identifying male-specific issues.


A cluster randomised trial of educational messages to improve the primary care of diabetes.
Regular laboratory test monitoring of patient parameters offers a route for improving the quality of chronic disease care. We evaluated the effects of brief educational messages attached to laboratory test reports on diabetes care. A programme of cluster randomised controlled trials was set in primary care practices in one primary care trust in England. Participants were the primary care practices' constituent healthcare professionals and patients with diabetes. Interventions comprised brief educational messages added to paper and electronic primary care practice laboratory test reports and introduced over two phases. Phase one messages, attached to Haemoglobin A1c (HbA1c) reports, targeted glycaemic and cholesterol control. Phase two messages, attached to albumin:creatinine ratio (ACR) reports, targeted blood pressure (BP) control, and foot inspection. Main outcome measures comprised practice mean HbA1c and cholesterol levels, diastolic and systolic BP, and proportions of patients having undergone foot inspections. Initially, 35 out of 37 eligible practices participated. Outcome data were available for a total of 8,690 patients with diabetes from 32 practices. The BP message produced a statistically significant reduction in diastolic BP (-0.62 mmHg; 95% confidence interval -0.82 to -0.42 mmHg) but not systolic BP (-0.06 mmHg, -0.42 to 0.30 mmHg) and increased the odds of achieving target BP control (odds ratio 1.05; 1.00, 1.10). The foot inspection message increased the likelihood of a recorded foot inspection (incidence rate ratio 1.26; 1.18 to 1.36). The glycaemic control message had no effect on mean HbA1c (increase 0.01%; -0.03 to 0.04) despite increasing the odds of a change in likelihood of HbA1c tests being ordered (OR 1.06; 1.01, 1.11). The cholesterol message had no effect (decrease 0.01 mmol/l, -0.04 to 0.05). Three out of four interventions improved intermediate outcomes or process of diabetes care. The diastolic BP reduction approximates to relative reductions in mortality of 3% to 5% in stroke and 3% to 4% in ischaemic heart disease over 10 years. The lack of effect for other outcomes may, in part, be explained by difficulties in bringing about further improvements beyond certain thresholds of clinical performance. Current Controlled Trials, ISRCTN2186314.


Correlating Eucalyptus leaf metabolomics with preference of the bronze bug, Thaumastocoris peregrinus.
Eucalyptus species are among the most planted trees in forestry production, an ever-increasing commercial activity worldwide. Forestry expansion demands a continuous search for preventive and sanitary measures against pests and diseases. Massive application of phytosanitary products is incompatible with the forestry sector, so forest health management must be based on other principles. In this context, studies on insect plant relationships mediated by plant metabolites may contribute information relevant to plant resistance and genotype selection. In this study, we analyzed the leaf metabolome of four Eucalyptus species commonly planted in southern South America, to correlate this chemical information with feeding preference of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae), an important pest of eucalypt plantations. Gas chromatography mass spectrometry analyses were performed on polar and non-polar leaf extracts from Eucalyptus globulus, Eucalyptus grandis, Eucalyptus robusta, and Eucalyptus tereticornis (Myrtaceae). Feeding preferences were assessed in two-choice laboratory bioassays resulting in a preference gradient of the four plant species. Moreover, a performance bioassay where we contrasted survival and development time between the most and least preferred plants, showed a clear correlation with preference both in survival and developmental time of the most susceptible nymph instar. We found that species with high or low feeding preferences differ significantly in several foliar metabolites, which may be acting as feeding stimulants or deterrents for T. peregrinus. These findings may provide useful criteria for choosing Eucalyptus genotypes when planting in bronze bug infested areas.


Convenient Access to Professional Interpreters in the Hospital Decreases Readmission Rates and Estimated Hospital Expenditures for Patients With Limited English Proficiency.
Twenty-five million people in the United States have limited English proficiency (LEP); this growing and aging population experiences worse outcomes when hospitalized. Federal requirements that hospitals provide language access services are very challenging to implement in the fast-paced, 24-hour hospital environment. To determine if increasing access to professional interpreters improves hospital outcomes for older patients with LEP. Natural experiment on a medicine floor of an academic hospital. Patients age 50 years or above discharged between January 15, 2007 and January 15, 2010. Dual-handset interpreter telephone at every bedside July 15, 2008 to Mar 14, 2009. Thirty-day readmission, length of stay, estimated hospital expenditures. Of 8077 discharges, 1963 were for LEP, and 6114 for English proficient patients. There was a significant decrease in observed 30-day readmission rates for the LEP group during the 8-month intervention period compared with 18 months preintervention (17.8% vs. 13.4%); at the same time English proficient readmission rates increased (16.7% vs. 19.7%); results remained significant in adjusted analyses. This improved readmission outcome for the LEP group was not maintained during the subsequent postintervention period when the telephones became less accessible. There was no significant intervention impact on length of stay in either unadjusted or adjusted analyses. After accounting for interpreter services costs, the estimated 119 readmissions averted during the intervention period were associated with estimated monthly hospital expenditure savings of $161,404. Comprehensive language access represents an important, high value service that all medical centers should provide to achieve equitable, quality healthcare for vulnerable LEP populations.


125I-fibrinogen uptake following phlebography of the leg. Comparison of ionic and nonionic contrast media.
The incidence of venous thrombosis diagnosed with the 125I-fibrinogen uptake test was investigated in a prospective, randomized, double-blind study with the patient serving as his own control, using an ionic contrast medium (methylglucamine iothalamate 280) in one leg and a nonionic medium (metrizamide) in the other. The incidence of thrombosis was significantly lower with metrizamide (7%) than with methylglucamine iothalamate (28.6%). The authors conclude that metrizamide is the contrast medium of choice for phlebography.


[Computer-controlled maintenance of the dissolved oxygen level in a medium during controlled cultivation of Neisseria meningitidis].
A new method for controlling the processes of N. meningitidis cultivation by the content of oxygen dissolved in the medium with the use of a computer, including the software, technical and methodological provision of the computer-operated control system, has been developed. The processes of the cultivation of N. meningitidis, groups A and B, under fully controlled conditions have been realized. The computer-operated control of the content of oxygen in the medium, the most important parameter of the process, has been shown to be essentially more effective than manual control. Information, accumulated in the course of the experiment, may be used in future for correction of mathematical models and optimization of the process. This will by necessary for obtaining biomass with predetermined properties in the development of new diagnostic and vaccine preparations.


Palliative dialysis: a change of perspective.
The aging phenomenon of dialysis patients is a worldwide reality, observed in developed and developing countries. Those patients have high incidence of chronic conditions along with high mortality rates and for some of them a decline in functional status within the first 12 months of dialysis therapy. Nevertheless, the elderly dialysis patients represent a very heterogeneous group where prognostic tools may help the decision-making process together with family members, medical staff and the patients. Despite the fact that there are many validated prognostic tools in elderly population, no score has the aim to guide the decision to withhold or withdrawn the dialysis procedure; therefore, in many cases, a time-limited trial is supported. After the failure of improvement in life quality and certitude of the poor prognosis, the withdrawing from renal replacement therapy can be done. Medical literature, from developed countries, brings robust evidence that the process of withdrawing the dialysis procedure, after a fail in the so-called "time-limited trial", along with good quality palliative care in this scenario is related to a good quality of death. We, on the other hand, believe that the withdrawing process in countries where hospice and good palliative care is not a reality may be associated with bad outcomes. Therefore, this review discusses a way to improve end-of-life symptoms in countries where palliative care facilities are not a reality, the so-called "palliative dialysis".


[Immunological studies of the cerebrospinal fluid in patients with amyotrophic lateral sclerosis].
Immunochemistry was used to measure the concentrations of immunoglobulins and some individual proteins in the CSF of patients with lateral amyotrophic sclerosis. The data obtained did not show any well-defined deviations from the reference intervals of the study parameters. It was impossible either to catch any changes during routine examination of the CSF. No characteristic features were revealed depending on the patients' clinical status. The study parameters do not provide anything definite for the pathogenesis and diagnosis of lateral amyotrophic sclerosis. The assessment of the results indicates that immunologically, the disease is more distant from the other demyelinating diseases.


Structural insights into novel mechanisms of inhibition of the major β-carbonic anhydrase CafB from the pathogenic fungus Aspergillus fumigatus.
In fungi the β-class of carbonic anhydrases (β-CAs) are zinc metalloenzymes that are essential for growth, survival, differentiation, and virulence. Aspergillus fumigatus is the most important pathogen responsible for invasive aspergillosis and possesses two major β-CAs, CafA and CafB. Recently we reported the biochemical characterization and 1.8 Å crystal structure of CafA. Here, we report a crystallographic analysis of CafB revealing the mechanism of enzyme catalysis and establish the relationship of this enzyme to other β-CAs. While CafA has a typical open conformation, CafB, when exposed to acidic pH and/or an oxidative environment, has a novel type of active site in which a disulfide bond is formed between two zinc-ligating cysteines, expelling the zinc ion and stabilizing the inactive form of the enzyme. Based on the structural data, we generated an oxidation-resistant mutant (Y159A) of CafB. The crystal structure of the mutant under reducing conditions retains a catalytic zinc at the expected position, tetrahedrally coordinated by three residues (C57, H113 and C116) and an aspartic acid (D59), and replacing the zinc-bound water molecule in the closed form. Furthermore, the active site of CafB crystals grown under zinc-limiting conditions has a novel conformation in which the solvent-exposed catalytic cysteine (C116) is flipped out of the metal coordination sphere, facilitating release of the zinc ion. Taken together, our results suggest that A. fumigatus use sophisticated activity-inhibiting strategies to enhance its survival during infection.


Non-gyrA-mediated ciprofloxacin resistance in laboratory mutants of Streptococcus pneumoniae.
Two strains of Streptococcus pneumoniae, M4 (NCTC 7465, type strain) and M5 (clinical isolate), and their respective ciprofloxacin-resistant mutants, M4/C1, M5/C1 and M5/C3, were evaluated. All mutants were stable after one year's storage and all grew more slowly in Brain Heart Infusion broth than the parent. The MICs of ciprofloxacin, sparfloxacin and tosufloxacin were increased for the mutants of M4, whereas the mutants of M5 were less susceptible to ciprofloxacin only. The optimal bactericidal concentration (OBC) of each quinolone for all the strains was approximately ten-fold greater than the MIC. The OBCs for the mutants were increased for ciprofloxacin, but not for the other two quinolones. The DNA synthesis IC50 values of all quinolones correlated well with the MIC of each drug. All quinolones accumulated rapidly within all five strains; 10 mM magnesium chloride decreased the concentration of quinolone accumulated, but carbonyl cyanide m-chlorophenyl hydrazone had no effect. Mutant strains M4/C1, M5/C1 and M5/C3 accumulated less quinolone than their respective parent strains. DNA sequencing of those regions of gyrA and gyrB corresponding to the quinolone resistance-determining region in other bacteria did not reveal any differences between the parent and mutant strains.


[Effectiveness of ethacizin in ventricular arrhythmias refractory to prior anti-arrhythmia drug therapy].
Using Holter ECG monitoring, the efficiency of the new drug ethacizin was assessed on an individual basis in 48 patients with ventricular arrhythmias refractory to past treatments (group 1) and in 46 patients who responded to at least one of the available antiarrhythmic drugs (group 2). Ethacizin proved effective in 63% of first-group patients and in all second-group patients, with an overall response rate of 81%. The efficiency of ethacizin, ethmozin, and disopyramide was 100, 73 and 69%, respectively, in the same patients. It is suggested that separate assessments of new antiarrhythmic drugs are preferable in patients with treatment-resistant ventricular arrhythmias and those responding to at least one of the drugs available.


[Determination of T anb B lymphocytes in patients with rheumatic fever].
The populations of "T" and "B" lymphocytes were evaluated in the present work employing the rosette technique in patients with active rheumatic fever, diagnosed according to Jones' criterion. "B" lymphocytes were found diminished at 7 days after the acute episode of rheumatic fever and increased at 35 days. These findings can be correlated with the hypergammaglobulinemia observed in this type of cases. Although there were wide variations in the quantity of "T" lymphocytes in some patients with rheumatic fever in this study, the porcentual and absolute values of "T" lymphocytes had no modifications when these were compared with the control group. The patients showed leucocytosis when entered the hospital and 7 days after. At 35 days of hospitalization they presented discrete lymphocytosis. Since the patients with rheumatic fever usually receive anti-inflammatory treatment with steroids, and as these drugs are capable of decreasing selectively the "T" lymphocytes, their effect on the kinetics of the lymphocytes in rheumatic fever has to be elavuated in further studies.


Narrow lumbar spinal canal with "vascular" syndromes.
Symptoms suggestive of vascular origin, both venous and arterial, may be the presenting complaints in patients with lumbar spondylosis. Fourteen patients suspected of having vascular intermittent claudication were found to be free of vascular disease, but had cauda equina compromise from herniated disk, osteoarthritis, and hypertrophic ligaments. Complete follow-up data were available for seven patients. In three, claudication seemed typical; in four, atypical. At operation, herniated intervertebral disks, osteophytic bone, or hypertrophied ligamenta flava, or a combination, were found. All benefited from lumbar laminectomy. When patients with vascular-like symptoms are found to be free of arterial or venous disease, lumbar spondylosis (narrow lumbar canal syndrome) should be considered. Chronic incapacitation pain without vascular disease provides a clue, as does electromyography. Plain X-ray films of the lumbar spine do not show the abnormality; thus, myelography should be carried out even in the absence of neurologic signs.


N1 esophageal carcinoma: the importance of staging and downstaging.
To evaluate the effects of clinical staging and downstaging by induction chemoradiation therapy in patients with N1 esophageal carcinoma. Sixty-nine consecutive patients with regional lymph node metastases (cN1) according to clinical staging received induction therapy before surgery. These were compared to 75 patients both clinically and pathologically N1 (cN1/pN1) who underwent surgery without induction therapy and 79 patients clinically and pathologically not N1 (cN0/pN0) who underwent surgery without induction therapy. Analyses focused on survival and the cost and benefit of therapy. For comparison, the extremes of 5-year survival were 69% for cN0/pN0 patients who underwent surgery alone and 12% for cN1/pN1 patients who underwent surgery alone. Of 69 patients who received induction therapy, 37 were pN0 at resection (downstaged); they had an intermediate survival of 37% at 5 years. Those patients not downstaged with induction therapy had a 12% 5-year survival, similar to patients with cN1/pN1 who underwent surgery alone. After adjusting for the strongest predictors of poor outcome, pN1, and increasing N1 burden, a modest increased risk of death after induction therapy was identified. However, this cost of induction therapy was more than counterbalanced by the benefit of improved survival of downstaging to pN0. (1) pN1 is the strongest determinant of poor outcome. (2) cN1 patients who are downstaged by induction chemoradiation therapy to pN0 have an intermediate outcome. (3) cN1 patients who are not downstaged by induction therapy have a poor outcome.


In vitro and in vivo studies of a naturally occurring variant of the human p85alpha regulatory subunit of the phosphoinositide 3-kinase: inhibition of protein kinase B and relationships with type 2 diabetes, insulin secretion, glucose disappearance constant, and insulin sensitivity.
In humans, the Met326Ile missense variant of the p85alpha regulatory subunit of the phosphoinositide 3-kinase (PI3K) has been associated with either significant reductions in glucose effectiveness and intravenous glucose tolerance in Caucasians or a significantly higher insulin secretory response in Pima Indians. In the present study, we genotyped 1,190 Caucasian males to evaluate the impact in vivo of the Met326Ile variant of the p85alpha subunit of PI3K on the acute insulin response, intravenous glucose tolerance, insulin-mediated glucose uptake, and the prevalence of type 2 diabetes after 20 years of follow-up. We also expressed the variant in vitro to evaluate the impact on insulin-stimulated activation of protein kinase B (PKB). The Met326Ile variant of p85alpha was not associated with type 2 diabetes or with alterations in insulin secretion, insulin sensitivity, or intravenous glucose tolerance in vivo. Expressed in vitro, the Ile326 and the Met326 variant acted equally as a dominant-negative and prevented (60-70% inhibition) insulin-mediated activation of PKB by inhibiting the phosphorylation of PKB at Thr308. We conclude that the Met326Ile variant of the p85alpha regulatory subunit of PI3K is likely to be as functionally normal in vivo as in vitro.


Trigeminal orosensation and ingestive behavior in the rat.
A deafferentation procedure was used to examine the contributions of trigeminal orosensation to the control of ingestive behavior in the rat. The procedure removed somatosensory input from the mouth, sparing olfaction, lingual taste, and vibrissae inputs as well as proprioceptive afferents from and efferents to the jaw muscles. Rats with sections of tongue or jaw muscle efferents served as oromotor controls. Bilateral trigeminal orosensory deafferentation was followed by an array of effects on ingestive behavior whose magnitudes were proportional to the extent of the deafferentation. The "trigeminal syndrome" includes a disruption of food and water intake (aphagia, adipsia), impairments in the sensorimotor control of eating and drinking, decreased responsiveness to food and water, and a reduction in the level of body weight regulation. Trigeminal deafferentation spared elementary ingestive movement patterns (biting, licking, chewing) but disrupted their control by the perioral stimuli, which normally elicit them, so that eating and drinking sequences were either aborted or inefficient. Deficits in food intake varied with the sensory properties of the diet. Recovery of intake took place along a palatability gradient, and recovery of water intake paralleled that of dry food. The chronically reduced body weight was caused by a persistent hypophagia and reflects reduced responsiveness to food. These findings suggest a considerable degree of overlap in the neural mechanisms mediating the sensorimotor and motivational control of intake in the rat.


UAG readthrough in mammalian cells: effect of upstream and downstream stop codon contexts reveal different signals.
Translation termination is mediated through an interaction between the release factors eRF1 and eRF3 and the stop codon within its nucleotide context. Although it is well known that the nucleotide contexts both upstream and downstream of the stop codon, can modulate readthrough, little is known about the mechanisms involved. We have performed an in vivo analysis of translational readthrough in mouse cells in culture using a reporter system that allows the measurement of readthrough levels as low as 10(-4). We first quantified readthrough frequencies obtained with constructs carrying different codons (two Gln, two His and four Gly) immediately upstream of the stop codon. There was no effect of amino acid identity or codon frequency. However, an adenine in the -1 position was always associated with the highest readthrough levels while an uracil was always associated with the lowest readthrough levels. This could be due to an effect mediated either by the nucleotide itself or by the P-site tRNA. We then examined the importance of the downstream context using eight other constructs. No direct correlation between the +6 nucleotide and readthrough efficiency was observed. We conclude that, in mouse cells, the upstream and downstream stop codon contexts affect readthrough via different mechanisms, suggesting that complex interactions take place between the mRNA and the various components of the translation termination machinery. Comparison of our results with those previously obtained in plant cells and in yeast, strongly suggests that the mechanisms involved in stop codon recognition are conserved among eukaryotes.


Compensatory sagittal plane ankle gait mechanics: Are they present in patients with a weak or stiff hip?
Simulations suggest that subjects with reduced hip range of motion (ROM) and/or weakness can achieve more normal walking mechanics through compensations at the ankle. The aims of this study were to assess whether subjects with reduced hip ROM (Stiff hip) or hip flexor weakness (Weak hip) exhibit ankle compensations during walking and investigate redistribution of power in the lower extremity joints. Retrospective gait data were reviewed (IRB-approved hip registry). Preoperative kinematic/kinetic walking data were collected in patients with: adolescent hip dysplasia (AHD), femoral acetabular impingement (FAI), and Legg-Calvé Perthes disease (Perthes). AHD patients with significantly weak hip flexors on their affected side were included (Weak hip group). The Gait Profile Score (GPS) was calculated on the affected side of the FAI and Perthes groups to identify patients who had a Stiff hip. Patients who had undergone a hip arthrodesis (Fusion) were also included (Stiff hip group). Ankle kinematics/kinetics were compared to healthy participants (Control). The total positive work of sagittal plane hip, knee and ankle power were compared along with the distribution of power. Patients in the Weak/Stiff hip groups did not walk with greater ankle plantarflexion, peak push-off power or positive ankle work on their affected sides compared to Control. Ankle work contribution (percentage of total positive work) on the affected or unaffected sides was greater in the Perthes and Hip Fusion patients compared to Control. Significant gait abnormalities on the unaffected side were observed. Patients with a weak or stiff hip did exhibit altered ankle mechanics during walking. Greater percent ankle work contribution appeared to correspond with hip stiffness. In patients with hip pathology the redistribution of power among the lower extremity joints can highlight the importance of preserving ankle function.


Post-response βγ power predicts the degree of choice-based learning in internally guided decision-making.
Choosing an option increases a person's preference for that option. This phenomenon, called choice-based learning (CBL), has been investigated separately in the contexts of internally guided decision-making (IDM, e.g., preference judgment), for which no objectively correct answer exists, and externally guided decision making (EDM, e.g., perceptual decision making), for which one objectively correct answer exists. For the present study, we compared decision making of these two types to examine differences of underlying neural processes of CBL. As IDM and EDM tasks, occupation preference judgment and salary judgment were used, respectively. To compare CBL for the two types of decision making, we developed a novel measurement of CBL: decision consistency. When CBL occurs, decision consistency is higher in the last-half trials than in first-half trials. Electroencephalography (EEG) data have demonstrated that the change of decision consistency is positively correlated with the fronto-central beta-gamma power after response in the first-half trials for IDM, but not for EDM. Those results demonstrate for the first time the difference of CBL between IDM and EDM. The fronto-central beta-gamma power is expected to reflect a key process of CBL, specifically for IDM.


Prevalence of neural tube defects, maternal HIV status, and antiretroviral therapy from a hospital-based birth defect surveillance in Kampala, Uganda.
The estimated prevalence of neural tube defects (NTDs) in Africa is 11.7 per 10,000 live births; however, data on the impact of antiretroviral therapy (ART) during pregnancy and the risk for birth defects in Africa are limited. Data from a hospital-based surveillance program at four hospitals in Kampala, Uganda were used to estimate the baseline prevalence of NTDs and assess potential associations with HIV status and ART use. All live births, stillbirths, and spontaneous abortions delivered at the participating hospitals affected with selected birth defects between August 2015 and December 2018 were included. Trained midwives collected data from hospital records, maternal interviews, photographs, and narrative descriptions of birth defects. We estimated NTD prevalence per 10,000 births (live, stillbirths, spontaneous abortions), prevalence ratios, and 95% confidence intervals (CIs). A total of 110,752 births from 107,133 women were included in the analysis; 9,394 (8.8%) women were HIV-infected and among those with HIV infection, 95.6% (n = 8,977) were on ART at delivery. Overall, 109 births were affected with NTDs, giving a prevalence of 9.8 (95% CI [8.2, 11.9]). Spina bifida (n = 63) was the most common type of NTD, with a prevalence of 5.7 (95% CI [4.4, 7.3]), followed by anencephaly (n = 31), with a prevalence of 2.8 (95% CI [2.0, 4.0]). The prevalence of NTDs among births in Kampala, Uganda is consistent with current estimates for Africa. With the continued introduction of new medications that may be taken during pregnancy, sustainable birth defect surveillance systems and pharmacovigilance are indicated.


Diffuse reflectance spectroscopy of the human tympanic membrane in otitis media.
We have investigated if features in the diffuse reflectance spectra from in vivo spectroscopic measurements of the tympanic membrane could aid the diagnosis of otitis media in children. Diffuse reflectance spectroscopy, in the visible wavelength range, was used in 15 ears from children with otitis media with effusion before and after myringotomy and in 15 healthy ears as a reference. Two previously published erythema detection algorithms yielded numerical quantities of haemoglobin content. With a combination of the algorithms, induced erythema (after myringotomy) was distinguished from healthy ears using Student's t-test (p < 0.01). Otitis media with mucous effusion was distinguished from (1) otitis media with serous effusion, (2) induced erythema and (3) healthy ears, (p < 0.05) using Student's t-test for independent groups and the paired t-test for dependent groups. Our results imply that reflectance spectroscopy is a promising technique to be used for the diagnosis of otitis media.


Early information processing deficit in schizophrenia. New findings using schizophrenic subgroups and manic control subjects.
In recent years, the idea that schizophrenia involves a primary disturbance of the higher cognitive (ie, cortical) thinking processes has been challenged by investigators who have shown that there may be a primary disturbance in schizophrenia in the early stages of information processing that occurs during the first few hundred milliseconds after the stimulus reaches the sense organs. Among the hypothesized early information processing deficits are deficiencies in iconic storage (a brief peripheral memory store) and slowness of processing from iconic storage to a more permanent memory system. Three experiments were conducted using tachistoscopically presented stimuli in order to evaluate these two stages of information processing (iconic storage and speed of processing) in schizophrenic and control subjects. Results converged in supporting the hypothesis, that independent of iconic storage and sensory registration, slow information processing is a relatively stable deficit of schizophrenic patients with a poor prognosis. The schizophrenic patients with a good prognosis had a similar deficit, which was reversible. Results are discussed as they relate to the early information processing deficit theories of schizophrenia.


Anthraquinone distribution in the hypogeal apparatus of Rubia peregrina L. growing wild in Sardinia.
The isolation, characterisation and quantification of anthraquinones, from hypogeal apparatus of Rubia peregrina L. (Wild Madder) growing wild in Sardinia (Italy) are described. The present study allowed us to perform an easy and fast HPLC analytical method for the detection of anthraquinones in all underground parts of plant. Concentrations of these compounds are variable in the different hypogeal parts before and after hydrolysis of the crude methanolic extracts. Before hydrolysis, we found that pseudopurpurin was 0.18% in rhizomes, alizarin 0.02% in rhizomes and principal stolons, and purpurin 0.56% in stolons of second order. Rubiadin is the major constituent of R. peregrina and reached a higher concentration in principal stolons of up to 4.8%. After hydrolysis, we found some drastic changes: an increase of purpurin in rhizomes (0.04%) and pseudopurpurin in all hypogeal parts (around 0.57%), and a decrease of purpurin in all parts of hypogeal apparatus. The most important change was found in rubiadin concentration, which reached its maximum in principal stolons (11.3%). No anthraquinones were found in the aerial parts of the plant.


Characterization of a GH36 α-D-Galactosidase Associated with Assimilation of Gum Arabic in Bifidobacterium longum subsp. longum JCM7052.
We recently characterized a 3-O-α-D-galactosyl-α-L-arabinofuranosidase (GAfase) for the release of α-D-Gal-(1→3)-L-Ara from gum arabic arabinogalactan protein (AGP) in Bifidobacterium longum subsp. longum JCM7052. In the present study, we cloned and characterized a neighboring α-galactosidase gene (BLGA_00330; blAga3). It contained an Open Reading Frame of 2151-bp nucleotides encoding 716 amino acids with an estimated molecular mass of 79,587 Da. Recombinant BlAga3 released galactose from α-D-Gal-(1→3)-L-Ara, but not from intact gum arabic AGP, and a little from the related oligosaccharides. The enzyme also showed the activity toward blood group B liner trisaccharide. The specific activity for α-D-Gal-(1→3)-L-Ara was 4.27- and 2.10-fold higher than those for melibiose and raffinose, respectively. The optimal pH and temperature were 6.0 and 50 °C, respectively. BlAga3 is an intracellular α-galactosidase that cleaves α-D-Gal-(1→3)-L-Ara produced by GAfase; it is also responsible for a series of gum arabic AGP degradation in B. longum JCM7052.


Beyond climate envelopes: effects of weather on regional population trends in butterflies.
Although the effects of climate change on biodiversity are increasingly evident by the shifts in species ranges across taxonomical groups, the underlying mechanisms affecting individual species are still poorly understood. The power of climate envelopes to predict future ranges has been seriously questioned in recent studies. Amongst others, an improved understanding of the effects of current weather on population trends is required. We analysed the relation between butterfly abundance and the weather experienced during the life cycle for successive years using data collected within the framework of the Dutch Butterfly Monitoring Scheme for 40 species over a 15-year period and corresponding climate data. Both average and extreme temperature and precipitation events were identified, and multiple regression was applied to explain annual changes in population indices. Significant weather effects were obtained for 39 species, with the most frequent effects associated with temperature. However, positive density-dependence suggested climatic independent trends in at least 12 species. Validation of the short-term predictions revealed a good potential for climate-based predictions of population trends in 20 species. Nevertheless, data from the warm and dry year of 2003 indicate that negative effects of climatic extremes are generally underestimated for habitat specialists in drought-susceptible habitats, whereas generalists remain unaffected. Further climatic warming is expected to influence the trends of 13 species, leading to an improvement for nine species, but a continued decline in the majority of species. Expectations from climate envelope models overestimate the positive effects of climate change in northwestern Europe. Our results underline the challenge to include population trends in predicting range shifts in response to climate change.


Defects in methylthioadenosine phosphorylase are associated with but not responsible for methionine-dependent tumor cell growth.
A large proportion of human tumor-derived cell lines and primary tumor cells show methionine-dependent growth. This phenomenon refers to the ability of cells to grow in media containing methionine and the inability of cells to grow in media supplemented with methionine's precursor, homocysteine (Hcy). Methionine can be formed by two different pathways, the recycling pathway and the salvage pathway. To discover the basis for methionine-dependent growth, we have analyzed 12 tumor cell lines and 2 non-tumor-derived cell lines for defects in two key genes in different methionine synthetic pathways. We found little evidence that defects in methionine synthase expression or mutations in the MS gene are correlated with methionine-dependent growth. However, we did find a correlation between methionine-dependent growth and defects in expression of methylthioadenosine phosphorylase (MTAP), a key enzyme in the salvage pathway. Three of the four cell lines lacking detectable MTAP protein were unable to grow in Hcy-containing media, whereas all six of the MTAP-positive cell lines tested showed strong growth. However, when we introduced MTAP cDNA into MTAP-deficient MCF-7 cells, the resulting cell line was still defective in growth on Hcy, although it could now grow on the salvage pathway precursor methylthioadenosine. These findings indicate that salvage pathway defects are not causally related to methionine-dependent growth.


Membrane Sphingolipids Regulate the Fitness and Antifungal Protein Susceptibility of Neurospora crassa.
The membrane sphingolipid glucosylceramide (GlcCer) plays an important role in fungal fitness and adaptation to most diverse environments. Moreover, reported differences in the structure of GlcCer between fungi, plants and animals render this pathway a promising target for new generation therapeutics. Our knowledge about the GlcCer biosynthesis in fungi is mainly based on investigations of yeasts, whereas this pathway is less well characterized in molds. We therefore performed a detailed lipidomic profiling of GlcCer species present in Neurospora crassa and comprehensively show that the deletion of genes encoding enzymes involved in GlcCer biosynthesis affects growth, conidiation and stress response in this model fungus. Importantly, our study evidences that differences in the pathway intermediates and their functional role exist between N. crassa and other fungal species. We further investigated the role of GlcCer in the susceptibility of N. crassa toward two small cysteine-rich and cationic antimicrobial proteins (AMPs), PAF and PAFB, which originate from the filamentous ascomycete Penicillium chrysogenum. The interaction of these AMPs with the fungal plasma membrane is crucial for their antifungal toxicity. We found that GlcCer determines the susceptibility of N. crassa toward PAF, but not PAFB. A higher electrostatic affinity of PAFB than PAF to anionic membrane surfaces might explain the difference in their antifungal mode of action.


[Erroneous diagnosis 'neonatal hypoglycemia' due to incorrect preservation of blood samples].
The high frequency of neonatal hypoglycaemia found in the St. Catherine's Hospital, Eindhoven [corrected], (8.2% of all newborn infants had values of 1.8 mmol/l glucose or less) promoted us to test the effects of the different conditions of blood preservation before measuring in a consecutive series of 18 newborn infants at risk for hypoglycaemia. When blood was preserved in microtest tubes coated with sodium fluoride (NaF) and measured after 1 hour at room temperature, the glucose level decreased by 7 to 36%. Storage on melting ice for one hour prevented this decline completely. In our hospital sodium fluoride coated microtest tubes are used. They were found to contain various concentrations of NaF, ranging from 0.85 to 2.65 mg/ml. In a 2nd experiment with cord blood from 8 patients, we proved that a concentration of 16 mg/ml sodium fluoride was necessary to inhibit glycolysis and to prevent a fall of plasma glucose. The rapid determination of blood glucose values, i.e. within 10 minutes after the blood sample is taken, decreased, as expected, the frequency of hypoglycaemia in the newborn at risk.


Development and validation of GLVS (Generic Laparoscopic Video Scoring System), a tool for assessment of laparoscopic skills in gynaecology using videotaped procedures: Are edited videos representative of their full-length version?
Anonymized videotaped endoscopic procedures can be used for the assessment of surgical competence, but a reliable non-procedure-specific scoring system is needed for gynaecology. To design and evaluate the validity of the Generic Laparoscopic Video Scoring System (GLVS), a novel tool in the assessment of various gynaecological laparoscopic procedures. Seventeen anonymized unedited video recordings of various gynaecological laparoscopic procedures and the 4-minute-long edited versions of the same videos were independently scored by two experts, twice, using GLVS. Internal consistency reliability, test-retest, and inter-rater reliability of GLVS. We also compared the scored achieved by edited videos with those of the full-length version of the same videos. The mean score achieved by 4-minute-long edited videos was similar to that of the unedited version (p= 0.13 - 0.19). There was excellent correlation between the pooled scores for edited and unedited versions (intra-class correlation coefficient = 0.86). GLVS had excellent internal consistency reliability (Cronbach's alpha 0.92-0.97). Test-retest and inter-rater reliability were generally better for edited 4-minute-long videos compared to their full-length version. Test-retest reliability for edited videos was excellent for scorer 1 and good for scorer 2 with intra-class correlation coefficient (ICC) of 0.88 and 0.62 respectively. Inter-rater reliability was good for edited videos (ICC=0.64) but poor for full-length versions (ICC= -0.24). GLVS allows for objective surgical skills assessment using anonymized shortened self-edited videos of basic gynaecological laparoscopic procedures. Shortened video clips of procedures seem to be representative of their full-length version for the assessment of surgical skills. We devised and undertook a validation study for a novel tool to assess surgical skills using surgical video clips. We believe this addition clearly delineates the unique contributions of our study.


Incorporating radiation with anti-CD19 chimeric antigen receptor T-cell therapy for relapsed/refractory non-Hodgkin lymphoma: A multicenter consensus approach.
Anti-CD19 chimeric antigen receptor T-cell therapy (CART) has revolutionized the outcomes of relapsed and/or refractory B-cell non-Hodgkin lymphoma. However, CART is still limited by its availability, toxicity, and response durability. Not all patients make it to the CART infusion phase due to disease progression. Among those who receive CART, a significant number of patients experience life-threatening cytokine release syndrome toxicity, and less than half maintain a durable response with the majority relapsing in pre-existing sites of disease present pre-CART. Radiation therapy stands as a promising peri-CART and salvage treatment that can improve the outcomes of these patients. Evidence suggests that bridging radiotherapy prior to CART controls the disease during the manufacturing period, augments response rates and local control, cytoreduces/debulks the disease and decreases the severity of cytokine release syndrome, and may prolong disease-free intervals and survival especially in patients with bulky disease. Consolidative radiotherapy for residual post-CART disease alters the pattern of relapse and improves local recurrence-free and progression-free survivals. Salvage radiotherapy for relapsed post-CART disease has favorable survival outcomes when delivered comprehensively for patients with limited relapsed disease and palliates symptoms for patients with diffuse relapsed disease. The biology of the disease during the peri-CART period is poorly understood, and further studies investigating the optimal timing and dosing of radiation therapy (RT) are needed. In this review, we tackle the most significant challenges of CART, review and propose how RT can help mitigate these challenges, and provide The Mayo Clinic experts' approach on incorporating RT with CART.


[Adaptive response--the general biological tendency: facts, hipothesis, questions].
It is leading up to the proofs testifying about the general-biological nature of adaptive response (AR), which is similar unspecific defense effect: for example, small doses induce cytokines synthesis, providing the resistance to different infectious agents. The connection between AR, DNA-repair, anti-oxidative status, expression of TP53 gene is discussed. The absence of AR at some patients can be explained either individual sensitivity to challenging treatment or criteria of the estimation (chromosome aberration, apoptosis et. al.). These facts are showing that the absence of AR cannot be the indicator of the risk for health. So the idea about general biological existence of AR is postulated because it is a characteristic for different species.


[Effect of sodium hydroxybutyrate, phenamine, transamine, l-DOPA on the physical work capacity of animals under multiple stress conditions].
Complete restoration of performance capacity in rats following a preliminary single exertion is shown to occur against the background of action produced both by transamine and its combination with sodium oxybutyrate. After a preliminary 4-fold swimming transamine is effective only in combination with sodium oxybutyrate. With a combined action of sodium oxybutyrate and l-DOPA a prolonged increase of performance capacity of the animals under multiple loads is recorded.


A systematic review of interventions to increase attendance at health and fitness venues: identifying key behaviour change techniques.
Members' attendance at health and fitness venues typically declines over the course of their membership, with a likely negative impact on physical activity and health outcomes. This systematic review sought to examine the effectiveness of interventions to increase attendance at health and fitness venues and identify the behaviour change techniques (BCTs) included in effective interventions. A systematic search of seven databases was conducted. The Behaviour Change Technique Taxonomy was used to code the interventions. Cohen's d was used to assess the effectiveness of the interventions. Fourteen papers reporting 20 interventions were included in the review. Most interventions were found to have trivial or small effects on attendance, although one had a medium effect (d = 0.60) and three had a large effect (ds = 1.00, 1.37, 1.45). The interventions used a limited range of BCTs, with "Prompts/Cues" being the most frequently used. Of the interventions with large effect sizes, two used "Problem solving" and "Pros and cons" and one used "Goal setting (behaviour)" and "Review behaviour goals". Only a small number of studies have tested interventions to increase attendance at health and fitness venues, with predominantly trivial or small effects. With the possible exception of problem solving alongside decisional balance and goal setting alongside reviewing behaviour goals, there is little evidence for the effectiveness of specific BCTs. Further research is required to identify the key components of effective interventions to increase attendance at health and fitness venues.


Evaluation of the synthesis of sialic acid-PAMAM glycodendrimers without the use of sugar protecting groups, and the anti-HIV-1 properties of these compounds.
A study was undertaken to evaluate the feasibility of synthesizing six sialic acid-PAMAM glycodendrimers using unprotected sialic acid in as few as 1-4 steps using two different reaction pathways, and to assess the sulfated derivatives for anti-HIV activity. The syntheses were accomplished through either the direct attachment of the sialic acid carboxyl group to amine-terminated PAMAM (a divergent-like approach) using BOP coupling, or by first reacting sialic acid with a polar bifunctional spacer molecule, attaching the sugar-linker to carboxy-terminated PAMAM (a convergent-like approach), and again using BOP-mediated coupling reactions. It was hypothesized that the latter approach would be the most successful method, as any steric congestion between the sialic acid and the PAMAM would be minimized using an intervening polar linker. However, the divergent-like synthesis proved to be the superior method, resulting in 11.4%, 14%, and 28% of the fully substituted generations 0, 1, and 2 sialic acid-PAMAM conjugates, respectively, as compared to 6.4% of only the generation -0.5 sialic acid-linker-PAMAM conjugate for the convergent-like method. Upon sulfation of the four glycodendrimers, binding capabilities to the recombinant HIV protein, gp120, were assessed using an ELISA assay. Compounds that showed promising binding characteristics were then further assessed for inhibition of HIV-1 infection using a well-characterized luciferase reporter gene neutralization assay. The generation 2 sulfated sialic acid-PAMAM glycodendrimer, sulfo-6, bearing 16 sialic acids with 11 sulfate groups incorporated at 4.03% sulfur content by weight, was found to inhibit all four HIV-1 strains tested in the low micromolar range.


Safety of parathyroidectomy in older vs. younger patients with primary hyperparathyroidism.
Primary hyperparathyroidism is a prevalent endocrinopathy for which surgery is the only curative option. Parathyroidectomy is primarily recommended in younger and symptomatic patients, while there are still concerns regarding surgical complications in older patients. We therefore assessed the association of age with surgical outcomes in patients undergoing parathyroidectomy in a large population in Switzerland. Population-based cohort study of adult patients with primary hyperparathyroidism undergoing parathyroidectomy in Switzerland between 2012 and 2018. The cohort was divided into four age groups (<50 years, 50-64 years, 65-74 years, ≥75 years). The primary outcome was a composite of in-hospital postoperative complications. Secondary outcomes were intensive care unit (ICU) admission, unplanned 30-day-readmission, and prolonged length of hospital stay. We studied 2642 patients with a median (IQR) age of 62 (53-71) years. Overall, 111 patients had complications including surgical re-intervention, hypocalcemia, and vocal cord paresis. As compared to <50 year-old patients, older patients had no increased risk for in-hospital complications after surgery (50-64 years: odds ratio (OR): 0.51 (95% CI, 0.28 to 0.92); 65-74 years: OR: 0.72 (95% CI, 0.39 to 1.33); ≥75 years: OR: 1.03 (95% CI, 0.54 to 1.95), respectively. There was also no association of age and rates of ICU-admission and unplanned 30-day-readmission, but oldest patients had longer hospital stays (OR: 2.38 (95% CI, 1.57 to 3.60)). ≥50 year-old patients undergoing parathyroidectomy had comparable risk of in-hospital complications as compared with younger ones. These data support parathyroidectomy in even older patients with primary hyperparathyroidism as performed in clinical routine.


New perspectives on vitamin D.
Our perception of vitamin D has evolved since the early part of this century. As an active, dihydroxylated metabolite vitamin D is acknowledged today not as an essential dietary factor but rather as a major calciotropic hormone. This paper summarizes current understanding of the formation and action of vitamin D in health and disease. The importance of laboratory assessment of serum vitamin D metabolites in the diagnosis and management of disorders of calcium and phosphorus metabolism is discussed.


Neural-Network-Based Approaches for Optimization of Machining Parameters Using Small Dataset.
Surface quality is one of the most important indicators of the quality of machined parts. The analytical method of defining the arithmetic mean roughness is not applied in practice due to its complexity and empirical models are applied only for certain values of machining parameters. This paper presents the design and development of artificial neural networks (ANNs) for the prediction of the arithmetic mean roughness, which is one of the most common surface roughness parameters. The dataset used for ANN development were obtained experimentally by machining AA7075 aluminum alloy under various machining conditions. With four factors, each having three levels, the full factorial design considers a total of 81 experiments that have to be carried out. Using input factor-level settings and adopting the Taguchi method, the experiments were reduced from 81 runs to 27 runs through an orthogonal design. In this study we aimed to check how reliable the results of artificial neural networks were when obtained based on a small input-output dataset, as in the case of applying the Taguchi methodology of planning a four-factor and three-level experiment, in which 27 trials were conducted. Furthermore, this paper considers the optimization of machining parameters for minimizing surface roughness in machining AA7075 aluminum alloy. The results show that ANNs can be successfully trained with small data and used to predict the arithmetic mean roughness. The best results were achieved by backpropagation multilayer feedforward neural networks using the BR algorithm for training.


Innate immune responses reverse HIV cognitive disease in mice: Profile by RNAseq in the brain.
Antiretroviral therapy controls immunodeficiency in people with HIV but many develop mild neurocognitive disorder. Here we investigated HIV brain disease by infecting mice with the chimeric HIV, EcoHIV, and probing changes in brain gene expression during infection and reversal with polyinosinic-polycytidylic acid (poly I:C). EcoHIV-infected C57BL/6 mice were treated with poly I:C and monitored by assay of learning in radial arm water maze, RNAseq of striatum, and QPCR of virus burden and brain transcripts. Poly I:C reversed EcoHIV-associated cognitive impairment and reduced virus burden. Major pathways downregulated by infection involved neuronal function, these transcriptional changes were normalized by poly I:C treatment. Innate immune responses were the major pathways induced in EcoHIV-infected, poly I:C treated mice. Our findings provide a framework to identify brain cell genes dysregulated by HIV infection and identify a set of innate immune response genes that can block systemic infection and its associated dysfunction in the brain.


[Comparative studies of the diagnostic value of various parameters of Doppler frequency spectral analysis of the carotid artery].
Angiographic findings in 94 internal carotid arteries were compared to different parameters of Doppler frequency spectral analysis: systolic and late diastolic peak frequency of the internal carotid artery, ratio of the internal and common carotid systolic and late diastolic peak frequency, spectral broadening as calculated by subtracting internal carotid mean frequency from systolic or late diastolic peak frequency respectively. The normal values were calculated as the 97.5% confidence interval of these parameters in angiographically normal vessels. Specificity, sensitivity, overall accuracy and negative as well as positive predictive value were also calculated. The diagnostic value of the systolic peak frequency of the internal carotid artery proved not to be exceeded by any of the other parameters.


A miR-200c/141-BMI1 autoregulatory loop regulates oncogenic activity of BMI1 in cancer cells.
MicroRNAs (miRNAs) are known to function as oncomiRs or tumor suppressors and are important noncoding RNA regulators of oncogenesis. The miR-200c/141 locus on chromosome 12 encodes miR-200c and miR-141, two members of the miR-200 family, which have been shown to function as tumor suppressive miRNAs by targeting multiple oncogenic factors such as polycomb group protein BMI1. Here, we show that BMI1 reciprocally functions as a transcriptional repressor of the miR-200c/141 cluster and that BMI1 inhibitors upregulate expression of miR-200c and miR-141. Our data suggest that BMI1 binds to the miR-200c/141 promoter and regulates it through transcription factor binding motifs E-box 2 and Z-box 1 to repress expression of miR-200c/141 cluster. We also show that PTC-209, a small molecule inhibitor of BMI1 gene expression induces cellular senescence and transcriptionally upregulates expression of miR-200c/141 cluster in breast cancer cells. Furthermore, inhibition of expression of miR-200c or miR-141 overcomes tumor suppressive effects of PTC-209 including induction of cellular senescence and downregulation of breast cancer stem cell phenotype. Therefore, our studies suggest a reciprocal regulation between BMI1 and miR-200c/141 cluster, and that BMI1 inhibitory drugs can further amplify their inhibitory effects on BMI1 via multiple mechanisms including posttranscriptional regulation by upregulating BMI1 targeting miRNAs.


Osteocyte-derived sclerostin inhibits bone formation: its role in bone morphogenetic protein and Wnt signaling.
Sclerosteosis and Van Buchem disease are rare, high-bone-mass disorders that have been linked to deficiency in the SOST gene, encoding sclerostin. Sclerostin belongs to the DAN family of glycoproteins, of which multiple family members have been shown to antagonize bone morphogenetic protein (BMP) and/or Wnt activity. Sclerostin is specifically expressed by osteocytes and inhibits BMP-induced osteoblast differentiation and ectopic bone formation. Sclerostin binds only weakly to BMPs and does not inhibit direct BMP-induced responses. Instead, sclerostin antagonizes canonical Wnt signaling by binding to Wnt coreceptors, low-density lipoprotein receptor-related protein 5 and 6. Several lipoprotein receptor-related protein-5 mutants that cause the high-bone-mass trait are defective in sclerostin binding. Thus, high bone mass in sclerosteosis and Van Buchem disease may result from increased Wnt signaling due to the absence of or insensitivity to sclerostin.


Reversed-phase High-performance Liquid Chromatography Method for Analysis of Curcuminoids and Curcuminoid-loaded Liposome Formulation.
A simple, sensitive, precise and specific method for the determination of curcuminoids and curcuminoid-loaded liposome formulation was developed using reverse-phase high-performance liquid chromatography method. The analysis was performed isocratically on Zorbax Eclipse XDB-C18 column (150×4 mm, 5 μm), analytical column using UV detector and mobile phase consisting of 0.1% orthophosphoric acid and acetonitrile. The proposed method for curcuminoids was validated for linearity in the range from 50 to 300 ΅g/ml with correlation coefficient above 0.997. Intraday and interday precision studies showed the relative standard deviation less than 2%. The limit of detection and limit of quantitation values were 2.5 and 8.25 ΅g/ml, respectively. Forced degradation study for curcuminoids and liposomal curcuminoids sample was carried out and observed that proposed method was also suitable for finding degradation products in the sample. Proposed method was successfully applied to estimate curcuminoids content without any interference of other excipients from liposomal formulation. Therefore, the method developed is well suited for curcuminoids and its liposome estimation.


Putting Role 1 first: the Role 1 capability review.
To quantify the risk for delivering care at Role 1 in the Land Environment (point of wounding to hospital care) on current operations and set the conditions for systematic change to enhance future capability. UK, US and Danish Army Role 1 Subject Matter Experts (SMEs) (1) Questionnaire study ofUK SMEs to determine capability gaps; (2) Questionnaire study of US and Danish SMEs to benchmark UK capability; (3) Semi-structured interviews of UK SMEs; (4) In-theatre evaluation of deployed Role 1. Thirty two SMEs completed the questionnaire (68% response rate), comprising 25 medical officers (20 in clinical appointments; five in command and staff appointments), six nurses and one medical support officer. Results of the entire review were collated as a cross-Defence Lines of Development analysis, separating the specific experience of 1 Medical Regiment's Hybrid Foundation Training (HFT), Mission Specific Training (MST) and deployment cycle from the analysis gained from questionnaire studies, SME consultation and documentary evidence. The review generated 77 recommendations and 38 sub-recommendations. The top six messages of the review were (1) To balance the expressed desire to increase the ratio of trained Team Medics with the reality of generating credible instructors with clinical experience; (2) To recognise that inadequate experience for Combat Medical Technicians in Primary Healthcare in the Firm Base undermines their operational preparedness; (3) To recognise that Current Regimental Aid Post (RAP) at contingency without power lacks the rudimentary infrastructure of a modern Medical Treatment Facility; (4) To recognise that inappropriate deployment of personnel with chronic disease or acutely limiting conditions is a consistent trend for 20 years that highlights continuing system weaknesses in applying protective medical grading; (5) To accept that General Practitioner manning requires re-evaluating as an Operational Pinch Point, reviewing all options to maintain operational effectiveness including, but not focusing on, incentives; and (6) To recognise that a best practice template for Role 1 Healthcare Governance has been created that must endure.


Cough-Associated Changes in CSF Flow in Chiari I Malformation Evaluated by Real-Time MRI.
Invasive pressure studies have suggested that CSF flow across the foramen magnum may transiently decrease after coughing in patients with symptomatic Chiari I malformation. The purpose of this exploratory study was to demonstrate this phenomenon noninvasively by assessing CSF flow response to coughing in symptomatic patients with Chiari I malformation by using MR pencil beam imaging and to compare the response with that in healthy participants. Eight symptomatic patients with Chiari I malformation and 6 healthy participants were studied by using MR pencil beam imaging with a temporal resolution of ∼50 ms. Patients and healthy participants were scanned for 90 seconds (without cardiac gating) to continuously record cardiac cycle-related CSF flow waveforms in real-time during resting, coughing, and postcoughing periods. CSF flow waveform amplitude, CSF stroke volume, and CSF flow rate (CSF Flow Rate = CSF Stroke Volume × Heart Rate) in the resting and immediate postcoughing periods were determined and compared between patients and healthy participants. There was no significant difference in CSF flow waveform amplitude, CSF stroke volume, and the CSF flow rate between patients with Chiari I malformation and healthy participants during rest. However, immediately after coughing, a significant decrease in CSF flow waveform amplitude (P < .001), CSF stroke volume (P = .001), and CSF flow rate (P = .001) was observed in patients with Chiari I malformation but not in the healthy participants. Real-time MR imaging noninvasively showed a transient decrease in CSF flow across the foramen magnum after coughing in symptomatic patients with Chiari I malformation, a phenomenon not seen in healthy participants. Our results provide preliminary evidence that the physiology-based imaging method used here has the potential to be an objective clinical test to differentiate symptomatic from asymptomatic patients with Chiari I malformation.


Overuse injuries in the knee, back and hip of top elite female alpine skiers during the off-season preparation period: prevalence, severity and their association with traumatic preinjuries and training load.
To investigate knee, back and hip overuse injuries in top female elite alpine skiers during the off-season preparation, which has so far received little attention, and to assess their relationship with traumatic preinjuries and overall training load. A cohort of 26 females of the Swiss Alpine Ski-Team were prospectively surveyed by the use of the Oslo Sports Trauma Research Centre questionnaire. Along with the surveys also total training loads were monitored. Data on preinjuries were assessed retrospectively by baseline questionnaires and were verified by team medical records. 57.7% of the female skiers suffered from at least one severe traumatic knee injury during their preceding career. The average 2-weekly prevalence of overuse injuries was highest for the knee, followed by the back and hip. Technique specialists (major giant slalom and slalom) were more prone to back overuse injuries than speed specialists (major super-G and downhill). The occurrence of knee overuse complaints was directly associated to previous severe traumatic knee injuries and athletes' total training hours. In top female elite alpine skiers, knee, back and hip overuse injuries are relatively frequent. Moreover, discipline preferences, previous traumatic injuries and the overall training load may play an important role for their manifestation.


Chronic CaMKII inhibition blunts the cardiac contractile response to exercise training.
Activation of the multifunctional Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) plays a critical role modulating cardiac function in both health and disease. Here, we determined the effect of chronic CaMKII inhibition during an exercise training program in healthy mice. CaMKII was inhibited by KN-93 injections. Mice were randomized to the following groups: sham sedentary, sham exercise, KN-93 sedentary, and KN-93 exercise. Cardiorespiratory function was evaluated by ergospirometry during treadmill running, echocardiography, and cardiomyocyte fractional shortening and calcium handling. The results revealed that KN-93 alone had no effect on exercise capacity or fractional shortening. In sham animals, exercise training increased maximal oxygen uptake by 8% (p < 0.05) compared to a 22% (p < 0.05) increase after exercise in KN-93 treated mice (group difference p < 0.01). In contrast, in vivo fractional shortening evaluated by echocardiography improved after exercise in sham animals only: from 25 to 32% (p < 0.02). In inactive mice, KN-93 reduced rates of diastolic cardiomyocyte re-lengthening (by 25%, p < 0.05) as well as Ca(2+) transient decay (by 16%, p < 0.05), whereas no such effect was observed after exercise training. KN-93 blunted exercise training response on cardiomyocyte fractional shortening (63% sham vs. 18% KN-93; p < 0.01 and p < 0.05, respectively). These effects could not be solely explained by the Ca(2+) transient amplitude, as KN-93 reduced it by 20% (p < 0.05) and response to exercise training was equal (64% sham and 47% KN-93; both p < 0.01). We concluded that chronic CaMKII inhibition increased time to 50% re-lengthening which were recovered by exercise training, but paradoxically led to a greater increase in maximal oxygen uptake compared to sham mice. Thus, the effect of chronic CaMKII inhibition is multifaceted and of a complex nature.


Whole-brain mapping of venous vessel size in humans using the hypercapnia-induced BOLD effect.
Measuring the morphology of the cerebral microvasculature by vessel-size imaging (VSI) is a promising approach for clinical applications, such as the characterization of tumor angiogenesis and stroke. Despite the great potential of VSI, this method has not yet found widespread use in practice due to the lack of experience in testing it on healthy humans. Since this limitation derives mainly from the need for an invasive injection of a contrast agent, this work explores the possibility to employ instead the easily accessible blood oxygenation level dependent (BOLD) effect for VSI of the venous microstructure. It is demonstrated that BOLD-VSI in humans can be realized by a hypercapnic challenge using a fast gradient-echo (GE) and spin-echo (SE) sequence at 7T. Reproducible maps of the mean venous vessel radius, based on the BOLD-induced changes in GE and SE relaxation rates, could be obtained within a scan time of 10min. Moreover, the method yields maps of venous blood volume and vessel density. Owing to its non-invasive character, BOLD-VSI provides a low-risk method to analyze the venous microstructure, which will not only be useful in clinical applications, but also provide a better understanding of BOLD effect.


Serosal tissue: reactive tissue as a model for understanding mesotheliomas.
Serosal tissues consist of a surface mesothelial layer and subsurface spindled connective tissue cells. Surface cells are decorated with antibodies to both low and high molecular weight cytokeratin whereas subserosal cells only express the intermediate filament vimentin. Serosal injury results in the proliferation of multipotential subserosal cells (MSC) which have the ultrastructural morphology of myofibroblasts and yet co-express low molecular weight cytokeratin and vimentin. These cells appear responsible for the re-establishment of surface mesothelium during which they acquire high molecular weight cytokeratin and loose vimentin. There are many parallels between reactive and neoplastic serosal tissues. Desmoplastic/sarcomatoid mesotheliomas resemble the MSC and co-express low molecular weight cytokeratin and vimentin and epithelial mesotheliomas resemble surface mesothelium and express both low and high molecular weight cytokeratin. The ability of normal serosal tissue to modulate its cell shape and intermediate filament expression helps understand the diversity of serosal tumors.


TaASR1, a transcription factor gene in wheat, confers drought stress tolerance in transgenic tobacco.
Abscisic acid (ABA)-, stress-, and ripening-induced (ASR) proteins are reported to be involved in abiotic stresses. However, it is not known whether ASR genes confer drought stress tolerance by utilizing the antioxidant system. In this study, a wheat ASR gene, TaASR1, was cloned and characterized. TaASR1 transcripts increased after treatments with PEG6000, ABA and H(2)O(2). Overexpression of TaASR1 in tobacco resulted in increased drought/osmotic tolerance, which was demonstrated that transgenic lines had lesser malondialdehyde (MDA), ion leakage (IL) and reactive oxygen species (ROS), but higher relative water content (RWC) and superoxide dismutase (SOD) and catalase (CAT) activities than wild type (WT) under drought stress. Overexpression of TaASR1 in tobacco also enhanced the expression of ROS-related and stress-responsive genes under osmotic stress. In addition, transgenic lines exhibited improved tolerance to oxidative stress by retaining more effective antioxidant system. Finally, TaASR1 was localized in the cell nucleus and functioned as a transcriptional activator. Taken together, our results showed that TaASR1 functions as a positive factor under drought/osmotic stress, involved in the regulation of ROS homeostasis by activating antioxidant system and transcription of stress-associated genes.


SPBP is a sulforaphane induced transcriptional coactivator of NRF2 regulating expression of the autophagy receptor p62/SQSTM1.
Organisms exposed to oxidative stress respond by orchestrating a stress response to prevent further damage. Intracellular levels of antioxidant agents increase, and damaged components are removed by autophagy induction. The KEAP1-NRF2 signaling pathway is the main pathway responsible for cell defense against oxidative stress and for maintaining the cellular redox balance at physiological levels. Sulforaphane, an isothiocyanate derived from cruciferous vegetables, is a potent inducer of KEAP1-NRF2 signaling and antioxidant response element driven gene expression. In this study, we show that sulforaphane enhances the expression of the transcriptional coregulator SPBP. The expression curve peaks 6-8 hours post stimulation, and parallels the sulforaphane-induced expression of NRF2 and the autophagy receptor protein p62/SQSTM1. Reporter gene assays show that SPBP stimulates the expression of p62/SQSTM1 via ARE elements in the promoter region, and siRNA mediated knock down of SPBP significantly decreases the expression of p62/SQSTM1 and the formation of p62/SQSTM1 bodies in HeLa cells. Furthermore, SPBP siRNA reduces the sulforaphane induced expression of NRF2, and the expression of the autophagy marker protein LC3B. Both these proteins contain ARE-like elements in their promoter regions. Over-expressed SPBP and NRF2 acts synergistically on the p62/SQSTM1 promoter and colocalize in nuclear speckles in HeLa cells. Collectively, these results suggest that SPBP is a coactivator of NRF2, and hence may be important for securing enhanced and sustained expression of NRF2 induced genes such as proteins involved in selective autophagy.


Long-distance transport of hair lambs: effect of location in pot-belly trailers on thermo-physiology, welfare and meat quality.
Livestock transport exposes animals to a range of potential stressors that may compromise their welfare and final product quality, and those effects typically increase with the distance travelled. In North America, producers often use pot-belly vehicles for long hauls but little is known about their suitability for transporting lambs. We followed two long-distance trips using pot-belly trailers carrying 500 lambs (each) from Northern to Central Mexico in winter, to measure possible effects on animal welfare and meat quality. Sixty lambs per trip were placed at different locations within the pot-belly trailer. Animals were equipped with iButton Thermochron® temperature devices, which registered body temperature throughout pre- and post-slaughter stages. Despite the rather cool winter conditions, lambs placed in the "belly" and "nose" compartments had higher body temperatures at loading, during transport and lairage and after slaughter (carcass temperature). Those lambs also had higher levels of plasma cortisol, glucose and creatine kinase (CK), and a higher neutrophil/lymphocyte ratio. Regarding meat quality, ultimate pH (24 h) was higher in all locations, especially in LT4 location. Overall, the results suggest a link between thermal stress during transport, elevated physiological indicators of stress and poorer meat quality.