Evaluation of ischemia after ESWL: detection of free oxygen radical scavenger enzymes in renal parenchyma subjected to high-energy shock waves.
To evaluate the degree of ischemic formation at the tissue level after high-energy shock wave (HESW) application in a time- and dose-dependent manner, an experimental study in rabbits was performed. Following varying numbers of HESWs (1,000-2,000-3,000 shock waves) under fluoroscopic localization, treated kidneys were removed, by assessing tissue concentrations of two different free-radical scavenger enzymes (superoxide dismutase and catalase) the degree of formation of free oxygen radicals (i.e. ischemia) was evaluated. Evaluation of our results showed a statistically significant (p < 0.05) decrease in tissue scavenger enzyme levels (i.e. formation of free oxygen radicals) after 24 h following HESW application. However, results obtained 7 days after ESWL demonstrated disappearance of ischemia with normalization of tissue enzyme levels (p > 0.05).


Deprescribing medications for older adults in the primary care context: A mixed studies review.
This review investigates the factors that influence deprescribing of medications in primary care from the perspective of general practitioners (GPs) and community-living older adults. A mixed studies review structure was adopted searching Scopus, CINAHL, PsychINFO, ProQuest, and PubMed from January 2000 to December 2017. A manual search of reference lists was also conducted. Studies were included if they were original research available in English and explored general deprescribing rather than deprescribing of a specific class of medications. The Mixed Methods Assessment Tool was used to assess the quality of studies, and content analysis generated common categories across studies. Thirty-eight articles were included, and 7 key categories were identified. The review found that the factors that influence deprescribing are similar across and within health systems and mostly act as barriers. These factors remained unchanged across the review period. The structural organisation of health systems remains poorly suited to facilitate deprescribing. Individual knowledge gaps of both GPs and older adults influence practices and attitudes towards deprescribing, and significant communication gaps occur between GPs and specialists and between GPs and older adults. As a result, deprescribing decision making is characterised by uncertainty, and deprescribing is often considered only when medication problems have already arisen. Trust plays a complex role, acting as both a barrier and facilitator of deprescribing. Deprescribing is influenced by many factors. Despite recent interest, little change has occurred. Multilevel strategies aimed at reforming aspects of the health system and managing uncertainty at the practice and individual level, notably reducing knowledge limitations and closing communications gaps, may achieve change.


A High Throughput Approach to Reconstruct Partial-Body and Neutron Radiation Exposures on an Individual Basis.
Biodosimetry-based individualized reconstruction of complex irradiation scenarios (partial-body shielding and/or neutron + photon mixtures) can improve treatment decisions after mass-casualty radiation-related incidents. We used a high-throughput micronucleus assay with automated scanning and imaging software on ex-vivo irradiated human lymphocytes to: a) reconstruct partial-body and/or neutron exposure, and b) estimate separately the photon and neutron doses in a mixed exposure. The mechanistic background is that, compared with total-body photon irradiations, neutrons produce more heavily-damaged lymphocytes with multiple micronuclei/binucleated cell, whereas partial-body exposures produce fewer such lymphocytes. To utilize these differences for biodosimetry, we developed metrics that describe micronuclei distributions in binucleated cells and serve as predictors in machine learning or parametric analyses of the following scenarios: (A) Homogeneous gamma-irradiation, mimicking total-body exposures, vs. mixtures of irradiated blood with unirradiated blood, mimicking partial-body exposures. (B) X rays vs. various neutron + photon mixtures. The results showed high accuracies of scenario and dose reconstructions. Specifically, receiver operating characteristic curve areas (AUC) for sample classification by exposure type reached 0.931 and 0.916 in scenarios A and B, respectively. R2 for actual vs. reconstructed doses in these scenarios reached 0.87 and 0.77, respectively. These encouraging findings demonstrate a proof-of-principle for the proposed approach of high-throughput reconstruction of clinically-relevant complex radiation exposure scenarios.


Production of diacylglycerol by exogenous phospholipase C stimulates CTP:phosphocholine cytidylyltransferase activity and phosphatidylcholine synthesis in human neuroblastoma cells.
The involvement of endogenous diacylglycerol production in the stimulation of phosphatidylcholine synthesis by exogenous phospholipase C was examined using a neuroblastoma (LA-N-2) cell line. Phospholipase C treatment (0.1 unit/ml) of intact cells stimulated CTP:phosphocholine cytidylyltransferase activity significantly more effectively than did maximally effective concentrations of the synthetic diacylglycerol sn-1,2-dioctanoylglycerol (1 mM). When added to cells together with phospholipase C, oleic acid, but not dioctanoylglycerol, further increased cytidylyltransferase activity with respect to phospholipase C treatment alone, indicating that the enzyme was not maximally activated by the lipase. This suggests that the lack of additivity of diacylglycerol and phospholipase C reflects a common mechanism of action. The time course of activation of cytidylyltransferase by phospholipase C paralleled that of [3H]diacylglycerol production in cells prelabeled for 24 h with [3H]oleic acid. Diacylglycerol mass was similarly increased. Significant elevations of [3H]oleic acid and total fatty acids occurred later than did the increases in cytidylyltransferase activity and diacylglycerol levels. No significant reduction in total or [3H]phosphatidylcholine was elicited by this concentration of phospholipase C, but higher concentrations (0.5 unit/ml) significantly reduced phosphatidylcholine content. The stimulation of cytidylyltransferase activity by phospholipase C or dioctanoylglycerol was also associated with enhanced incorporation of [methyl-14C]choline into phosphatidylcholine. Dioctanoylglycerol was more effective than phospholipase C at stimulating the formation of [14C]phosphatidylcholine, and the effects of the two treatments were additive. However, further analysis revealed that dioctanoylglycerol served as a precursor for [14C]dioctanoylphosphatidylcholine as well as an activator of cytidylyltransferase; and when corrections were made for this effect, the apparent additivity disappeared. The results indicate that the generation of diacylglycerol by exogenous phospholipase C (and possibly the subsequent production of fatty acids via diacylglycerol metabolism) activates cytidylyltransferase activity in neuronal cells under conditions in which membrane phosphatidylcholine content is not measurably reduced.


Enhanced metabolic resistance mechanism endows resistance to metamifop in Echinochloa crus-galli (L.) P. Beauv.
Barnyardgrass (Echinochloa crus-galli (L.) P. Beauv.), one of the worst weeds in paddy fields in China, has been frequently reported evolving resistance to acetyl-CoA carboxylase (ACCase) inhibiting herbicides. However, in the previous research, more attention was paid to target-site resistance (TSR) mechanisms, the non-target-site resistance (NTSR) mechanisms have not been well-established. In this study, the potential mechanism of resistance in a metamifop-resistant E. crus-galli collected from Kunshan city, Jiangsu Province, China was investigated. Dose-response assays showed that the phenotypic resistant population (JS-R) has evolved 4.3-fold resistance to metamifop compared with the phenotypic susceptible population (YN-S). The ACCase CT gene sequencing and relative ACCase gene expression levels studies showed that no mutations were detected in the ACCase CT gene in both YN-S and JS-R, and there was no significant difference in the relative ACCase gene expression between YN-S and JS-R. After the pre-processing of glutathione-S-transferase (GSTs) inhibitor NBD-Cl, the resistance level of JS-R to metamifop was reversed 18.73%. Furthermore, the GSTs activity of JS-R plants was significantly enhanced compared to that of YN-S plants. UPLC-MS/MS revealed that JS-R plants had faster metabolic rates to metamifop than YN-S plants. Meanwhile, the JS-R popultion exhibited resistant to cyhalofop-butyl and penoxsulam. In summary, this study presented a novel discovery regarding the global emergence of metabolic resistance to metamifop in E. crus-galli. The low-level resistance observed in the JS-R population was not found to be related to TSR but rather appeared to be primarily associated with the overexpression of genes in the GSTs metabolic enzyme superfamily.


Objective interictal electrophysiology biomarkers optimize prediction of epilepsy surgery outcome.
Researchers have looked for rapidly- and objectively-measurable electrophysiology biomarkers that accurately localize the epileptogenic zone. Promising candidates include interictal high-frequency oscillation and phase-amplitude coupling. Investigators have independently created the toolboxes that compute the high-frequency oscillation rate and the severity of phase-amplitude coupling. This study of 135 patients determined what toolboxes and analytic approaches would optimally classify patients achieving post-operative seizure control. Four different detector toolboxes computed the rate of high-frequency oscillation at ≥80 Hz at intracranial EEG channels. Another toolbox calculated the modulation index reflecting the strength of phase-amplitude coupling between high-frequency oscillation and slow-wave at 3-4 Hz. We defined the completeness of resection of interictally-abnormal regions as the subtraction of high-frequency oscillation rate (or modulation index) averaged across all preserved sites from that averaged across all resected sites. We computed the outcome classification accuracy of the logistic regression-based standard model considering clinical, ictal intracranial EEG and neuroimaging variables alone. We then determined how well the incorporation of high-frequency oscillation/modulation index would improve the standard model mentioned above. To assess the anatomical variability across non-epileptic sites, we generated the normative atlas of detector-specific high-frequency oscillation and modulation index. Each atlas allowed us to compute the statistical deviation of high-frequency oscillation/modulation index from the non-epileptic mean. We determined whether the model accuracy would be improved by incorporating absolute or normalized high-frequency oscillation/modulation index as a biomarker assessing interictally-abnormal regions. We finally determined whether the model accuracy would be improved by selectively incorporating high-frequency oscillation verified to have high-frequency oscillatory components unattributable to a high-pass filtering effect. Ninety-five patients achieved successful seizure control, defined as International League against Epilepsy class 1 outcome. Multivariate logistic regression analysis demonstrated that complete resection of interictally-abnormal regions additively increased the chance of success. The model accuracy was further improved by incorporating z-score normalized high-frequency oscillation/modulation index or selective incorporation of verified high-frequency oscillation. The standard model had a classification accuracy of 0.75. Incorporation of normalized high-frequency oscillation/modulation index or verified high-frequency oscillation improved the classification accuracy up to 0.82. These outcome prediction models survived the cross-validation process and demonstrated an agreement between the model-based likelihood of success and the observed success on an individual basis. Interictal high-frequency oscillation and modulation index had a comparably additive utility in epilepsy presurgical evaluation. Our empirical data support the theoretical notion that the prediction of post-operative seizure outcomes can be optimized with the consideration of both interictal and ictal abnormalities.


Does rehearsal matter? Left anterior temporal alpha and theta band changes correlate with the beneficial effects of rehearsal on working memory.
Rehearsal during working memory (WM) maintenance is assumed to facilitate retrieval. Less is known about how rehearsal modulates WM delay activity. In the present study, 44 participants completed a Sternberg Task with either intact novel scenes or phase-scrambled scenes, which had similar color and spatial frequency but lacked semantic content. During the rehearsal condition participants generated a descriptive label during encoding and covertly rehearsed during the delay period. During the suppression condition participants did not generate a label during encoding and suppressed (repeated "the") during the delay period. This was easy in the former (novel scenes) but more difficult in the later condition (phase-scrambled scenes) where scenes lacked semantic content. Behavioral performance and EEG delay activity was analyzed as a function of maintenance strategy. Performance during WM revealed a benefit of rehearsal for phase-scrambled but not intact scenes. Examination of the absolute amplitude revealed three underlying sources of activity for rehearsal, including the left anterior temporal (ATL) and left and midline parietal regions. Increases in alpha and theta activity in ATL were correlated with improvement in performance on WM with rehearsal only when labeling was not automatic (e.g., phase-scrambled scenes), which may reflect differences in labeling and rehearsal (i.e., semantic associations vs. shallow labels). We conclude that rehearsal only benefits memory for visual stimuli that lack semantic information, and that this is correlated with changes in alpha and theta rhythms.


Local attributable burden disease to PM 2.5 ambient air pollution in Medellín, Colombia, 2010-2016.
Background: Exposure to 2.5-micron diameter air pollutants (PM 2.5) has been associated with an increased risk of illness and death worldwide; however, in Latin American health impacts assessment of this risk factor is scarce. Medellín is one of the most polluted cities in the region, with a population growth rate that is twice as high as that of other Colombian cities, which implies a growing population at risk. Methods: A descriptive study of the disease burden was carried out using the city as the unit of observation. Health events were selected based on epidemiologic evidence and the availability of the population attributable fraction associated with PM 2.5. The mortality records were taken from the module of deceased of the Single Registry of Affiliates of the Health System; the morbidity records were taken from the Individual Health Services Registries. For the estimation of the burden of disease, the current Global Burden of Disease guidelines were followed. Results: Attributable disability-adjusted life years to exposure to ambient PM 2.5 pollution (DALYs PM2.5) constituted 13.8% of total burden of the city. Males showed the greatest loss of DALYs PM2.5 due to acute events, while in women the greatest loss was due to chronic events. Ischemic heart disease, chronic diseases of the lower respiratory tract, and influenza and pneumonia were the events that contributed the most to DALYs PM2.5. 71.4% of the DALYs PM2.5 corresponded to mortality, mainly in the population over 65 years of age. Regarding attributable morbidity, acute events were more prevalent in both sexes, especially due to respiratory diseases Conclusion: Premature death among the elderly population has the greatest weight on burden of disease attributable to ambient PM 2.5 pollution, mainly due to respiratory and cardiovascular diseases, without significant differences according to gender.


Limited agreement between written and video asthma symptom questionnaires.
The prevalence of asthma remains difficult to determine with precision with no absolute or "gold" standard for diagnosis. A recently developed video questionnaire for epidemiological studies with less reliance on understanding written questions provides another tool for determining prevalence and severity of asthma. This report from the International Study of Asthma and Allergies in Childhood (ISAAC) examines the agreement between the ISAAC video questionnaires on respiratory symptoms and reported asthma. Between December 1993 and April 1995, 4952 children aged 13-14 years in two Canadian communities completed sequentially the ISAAC written and video questionnaires at school. The agreement between responses to the two questionnaires for reported wheeze ever, current wheeze, wheeze on exercise, and nocturnal wheeze (the latter three questions relating to symptoms in the last 12 months), and to any combination of the latter three questions was examined in the full sample and in those reporting diagnosed asthma, using concordance and kappa coefficients as measures of agreement. The prevalences of wheeze ever, current wheeze, wheeze on exercise, and nocturnal wheeze were significantly lower based on responses to the video questionnaire compared with the written questionnaire in both regions in the full sample and in those labeled as having asthma. Although concordance between video and written questionnaires always exceeded 60% and often exceeded 70% for related questions, agreement measured by the kappa statistic for each question was only fair to moderate (kappa = 0.22-0.51). We conclude that the video questionnaire yields lower reported prevalence rates for asthma symptoms, and that there is limited agreement between responses to the two questionnaires that is not explained by issues of language, culture, or literacy.


Implementing an inpatient rehabilitation Saturday service is associated with improved patient outcomes and facilitates patient flow across the health care continuum.
To evaluate the effectiveness of the introduction of a Saturday inpatient rehabilitation service in improving patient outcomes and facilitating hospital access and flow. A quasi-experimental study with a historical comparison group is presented. Data were evaluated for the 477 patients admitted and discharged in the 6 months following the implementation (October 2012-March 2013) of the service and 499 patients in the historical control group (April 2012-September 2012). Prior to the introduction of the service median number of patients admitted on a Saturday was 0 (range 0-3), post-implementation the median number of patients admitted on a Saturday increased to 2 (range 0-5), this difference was statistically significant (Z = -3.61, p < 0.001). Median regression modelling demonstrated that, after adjusting for admission Functional Independence (FIM) score, gender, length of stay (LOS) and age, there was a small but significant increase in discharge FIM scores (median increase 2.28, p = 0.027) for those patients admitted in the post-implementation phase. There was no such effect of the service on LOS. The implementation of the Saturday inpatient rehabilitation service was associated with improved patient outcomes and supported an increase in patient flow throughout the organization as evidenced by an increased number of Saturday patient admissions. Inpatient rehabilitation services do not operate in isolation; they are a critical link in the healthcare continuum. There is a mounting body of evidence that the introduction of weekend rehabilitation services has a positive impact on patient outcomes. To maximize health outcomes, access to critical services and patient flow, organizations and staff need to move away from the long-standing 5-day/week inpatient rehabilitation model and start transitioning to a model that incorporates weekend therapy services.


Extracellular matrix composition reveals complex and dynamic stromal-epithelial interactions in the mammary gland.
The mammary gland is an excellent model system to study the interplay between stroma and epithelial cells because of the gland's unique postnatal development and its distinct functional states. This review focuses on the contribution of the extracellular matrix (ECM) to stromal-epithelial interactions in the mammary gland. We describe how ECM physical properties, protein composition, and proteolytic state impact mammary gland architecture as well as provide instructive cues that influence the function of mammary epithelial cells during pubertal gland development and throughout adulthood. Further, based on recent proteomic analyses of mammary ECM, we describe known mammary ECM proteins and their potential functions, as well as describe several ECM proteins not previously recognized in this organ. ECM proteins are discussed in the context of the morphologically-distinct stromal subcompartments: the basal lamina, the intra- and interlobular stroma, and the fibrous connective tissue. Future studies aimed at in-depth qualitative and quantitative characterization of mammary ECM within these various subcompartments is required to better elucidate the function of ECM in normal as well as in pathological breast tissue.


Electrochemical studies of DNA immobilization onto the azide-terminated monolayers and its interaction with taxol.
A surface modification procedure for the creation of self-assembled monolayers (SAMs) that can be used as a scaffold for double-stranded DNA (dsDNA) incorporation onto the gold surfaces is described. The SAMs of an azidohexane thiol derivative were prepared on the Au electrode and then used for the immobilization of dsDNA. The electrochemical characteristics of dsDNA onto the SAM-modified gold electrode were investigated by cyclic voltammetry and electrochemical impedance spectroscopy, and the surface concentration of dsDNA onto the SAMs surface was estimated. The interaction of dsDNA with the anticancer drug, taxol (paclitaxel), was also studied on the surface of DNA/SAM/Au electrode. The observed decrease in the guanine oxidation peak current was used to monitor the interaction of taxol with DNA. The resulting Langmuir isotherm for taxol binding to DNA at the modified electrode was used to evaluate the binding constant of taxol-DNA. The results obtained supported the groove binding interaction of taxol with DNA. The modified electrode was used as a sensitive sensor for quantification of taxol in human serum sample.


Repeated lifestyle interventions lead to progressive weight loss: A retrospective review chart study.
This study aimed to investigate whether repeated lifestyle interventions lead to progressive weight loss or to weight cycling. A retrospective review chart study with follow-up on 2120 participants (mean±SD age 36±15 years; body weight 116±28 kg; fat 43±6%). All had participated in one to four 11-12 week lifestyle interventions (residential weight loss programme, mixed activities). Weight loss was promoted through a hypocaloric diet (-500 to -700 kcal/day) and daily physical activity (1-3 hours/day). Primary outcomes were weight loss and change in body composition (bioimpedance measurements) after the intervention periods and at follow-up. A total of 2120, 526, 139 and 47 people participated in one to four interventions with mean±SEM times from start to follow-up of 1.3±0.1, 2.9±0.2, 4.2±0.3 and 5.2±0.4 years respectively. Overall 50, 41, 18 and 11% of the participants were lost to follow-up after one to four interventions, respectively. The cumulated weight loss at follow-up increased with the number of interventions from one to four: 12.2±0.1, 15.9±0.7, 16.1±1.2 and 18.5±2.0 kg ( p<0.001). The ratios between cumulated loss of fat and fat free mass after one to four interventions decreased with the number of interventions (2.4, 2.2, 2.1 and 1.4). Rates of weight loss during the interventions ranged from 0.70±0.06 to 1.06±0.01 kg/week and the maximum weight regain during the follow-up periods was 0.039±0.007 kg/week. Repeated relatively short lifestyle interventions in a selected and motivated group can be an efficient method for weight loss maintenance with only limited body weight cycling in the interim periods. However, the relationship between loss of fat and fat free mass might change in an unfavourable direction.


Telling the truth: what do general practitioners say to patients with dementia or terminal cancer?
A questionnaire was sent to all general practitioners (GPs) in Cambridgeshire asking them about their practice in telling patients with dementia or terminal cancer their diagnosis. Although the majority of GPs would always or often tell patients of a diagnosis of terminal cancer, this was not the case in dementia; uncertainty of diagnosis was cited as the most important factor in not giving the diagnosis. The implications of these findings are discussed in the light of the GP's role in dementia and the development of new anti-Alzheimer's drugs.


Fate of [125I]insulin removed from the peritubular circulation of isolated perfused rat kidney.
Although there is considerable evidence that insulin is removed from the peritubular circulation of the mammalian kidney, it is unclear whether binding to insulin-specific receptors is involved in this process, whether after peritubular removal the hormone is degraded to small fragments with release into the circulation, or whether it merely undergoes a minor modification with loss of immunoreactivity. We examined the metabolism of [125I]insulin removed from the peritubular circulation of the nonfiltering isolated perfused rat kidney and compared it to that of [125I]insulin metabolized by filtering isolated kidneys and kidney homogenates. The results indicate that after peritubular removal, a small amount of insulin is degraded to form low-molecular-weight products similar to those seen with filtering kidneys and kidney homogenates. However, most of the insulin removed from the peritubular circulation is processed either to nonimmunoreactive products of molecular weight similar to that of insulin or, to a lesser extent, to products of larger molecular weight. Both these products are also formed by filtering kidneys. In the filtering kidney, the products having molecular weight similar to that of insulin probably originate from the peritubular process, because it is unlikely that material of this size could be derived from the filtration-absorption pathway. Of particular note was the finding that [125I]insulin trapped in the peritubular compartment of nonfiltering kidneys was displaced severalfold more effectively by unlabeled insulin than by several peptide hormones (P less than 0.01); the latter were no more effective than vehicle alone. The findings suggest the presence of peritubular insulin-specific receptors.


Instrumentation for small joint arthroscopy.
Proper instrumentation is basic to all podiatric surgical procedures, but in many instances one instrument can be substituted for another. In podiatric arthroscopy, however, the instrumentation is the "sine qua non" of the procedure; one instrument does not substitute for another and all have specific functions.


Short-term effects of cortisol implantation on blood biochemistry and thyroid hormones in previtellogenic great sturgeon Huso huso.
This study examined the effects of implanted cortisol on various aspects of intermediary metabolism of great sturgeon, Huso huso. Prior to experimentation all fish were examined using an endoscope to observe the stage of ovarian development. Subsequently, the 3-year-old female fish in the previtellogenic stage (mean body weight of 6759±53.2g) were intraperitoneally implanted with cocoa butter pellets containing cortisol to mimic the effects of chronic stress. The implant doses were 0 (C0; as control), 5 (C5) and 50 (C50) mg cortisol/kg body weight. Blood samples were taken every seven days during the four weeks of the experiment and analyzed for cortisol, glucose, adrenocorticotropic hormone (ACTH), total protein, total lipid, triiodothyronine (T3), thyroxine (T4), cholesterol and triglyceride content. Growth was reduced in all experimental groups and was not affected by cortisol treatment. Surprisingly, serum cortisol levels were higher in the C5 group than in the C50 throughout the experiment. A significant increase in glucose levels was observed in the cortisol-implanted fish from day 14 onwards. The high dose of cortisol elicited a significant increase in serum T3 and T4 levels. Fish implanted with the high cortisol dose also showed increases in serum ACTH, total lipid and cholesterol levels throughout a 28-day experimental period. The present study reveals that the negative effects of endoscopic surgery remain for at least four weeks and that a sustained-release implant of cortisol to mimic the effects of chronic stress affects metabolic responses. Since the adverse effects of endoscopic surgery on sturgeon welfare can be amplified by cortisol, special attention should be paid to the potential effects of chronic stress on sturgeon in culture.


The acute impact of ingestion of breads of varying composition on blood glucose, insulin and incretins following first and second meals.
Structural characteristics and baking conditions influence the metabolic responses to carbohydrate-containing foods. We hypothesized that consumption of whole wheat or sourdough breads would have a favourable effect on biomarkers of glucose homeostasis after first and second meals, compared with those for white bread. Ten overweight volunteers consumed 50 g available carbohydrate of each of the four breads (white, whole wheat, sourdough, whole wheat barley) followed 3 h later by a standard second meal. Blood was sampled for 3 h following bread ingestion and a further 2 h after the second meal for determination of glucose, insulin, paracetamol (indirect marker of gastric emptying), glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1). Glucose and GLP-1 responses to sourdough bread were lower (P < 0.05) than whole wheat and whole wheat barley breads. Glucose area under the curve (AUC) for sourdough bread was lower than those for whole wheat (P < 0.005) and whole wheat barley (P < 0.03) breads for the entire study. GIP AUC after sourdough bread ingestion was lower compared to white (P < 0.004) and whole wheat barley (P < 0.002) breads following the second meal. There were no significant differences in insulin and paracetamol concentrations among the test breads. Ultra-fine grind whole wheat breads did not result in postprandial responses that were lower than those of white bread, but sourdough bread resulted in lower glucose and GLP-1 responses compared to those of these whole wheat breads following both meals.


Structure and stability of the ankyrin domain of the Drosophila Notch receptor.
The Notch receptor contains a conserved ankyrin repeat domain that is required for Notch-mediated signal transduction. The ankyrin domain of Drosophila Notch contains six ankyrin sequence repeats previously identified as closely matching the ankyrin repeat consensus sequence, and a putative seventh C-terminal sequence repeat that exhibits lower similarity to the consensus sequence. To better understand the role of the Notch ankyrin domain in Notch-mediated signaling and to examine how structure is distributed among the seven ankyrin sequence repeats, we have determined the crystal structure of this domain to 2.0 angstroms resolution. The seventh, C-terminal, ankyrin sequence repeat adopts a regular ankyrin fold, but the first, N-terminal ankyrin repeat, which contains a 15-residue insertion, appears to be largely disordered. The structure reveals a substantial interface between ankyrin polypeptides, showing a high degree of shape and charge complementarity, which may be related to homotypic interactions suggested from indirect studies. However, the Notch ankyrin domain remains largely monomeric in solution, demonstrating that this interface alone is not sufficient to promote tight association. Using the structure, we have classified reported mutations within the Notch ankyrin domain that are known to disrupt signaling into those that affect buried residues and those restricted to surface residues. We show that the buried substitutions greatly decrease protein stability, whereas the surface substitutions have only a marginal affect on stability. The surface substitutions are thus likely to interfere with Notch signaling by disrupting specific Notch-effector interactions and map the sites of these interactions.


Is diabetes in Cushing's syndrome only a consequence of hypercortisolism?
Diabetes mellitus (DM) is one of the most frequent complications of Cushing's syndrome (CS). The aim of this study was to define the changes in insulin sensitivity and/or secretion in relation to glucose tolerance categories in newly diagnosed CS patients. Cross-sectional study on 140 patients with CS. A total of 113 women (80 with pituitary disease and 33 with adrenal disease, aged 41.7±15.7 years) and 27 men (19 with pituitary disease and eight with adrenal disease, aged 38.1±20.01 years) at diagnosis were divided according to glucose tolerance into normal glucose tolerance (CS/NGT), impaired fasting glucose and/or impaired glucose tolerance (CS/prediabetes), and diabetes (CS/DM) groups. Seventy-one patients had CS/NGT (49.3%), 26 (18.5%) had CS/prediabetes and 43 (30.8%) had CS/DM. Significant increasing trends in the prevalence of family history of diabetes (P<0.001), metabolic syndrome (P<0.001), age (P<0.001) and waist circumference (P=0.043) and decreasing trends in HOMA-β (P<0.001) and oral disposition index (DIo) (P<0.002) were observed among the groups. No significant trends in fasting insulin levels, area under the curve for insulin (AUCINS), Matsuda index of insulin sensitivity (ISI-Matsuda) and visceral adiposity index were detected. Impairment of glucose tolerance is characterized by the inability of β-cells to adequately compensate for insulin resistance through increased insulin secretion. Age, genetic predisposition and lifestyle, in combination with the duration and degree of hypercortisolism, strongly contribute to the impairment of glucose tolerance in patients with a natural history of CS. A careful phenotypic evaluation of glucose tolerance defects in patients with CS proves useful for the identification of those at a high risk of metabolic complications.


The use of smell identification tests in the diagnosis of Parkinson's disease in Brazil.
Smell identification tests may be of routine clinical value in the differential diagnosis of PD but are subject to cultural variation and have not been systematically evaluated in the Brazilian population. We have applied culturally adapted translations of the University of Pennsylvania 40-item smell identification test (UPSIT-40) and the 16-item identification test from Sniffin' Sticks (SS-16) to nondemented Brazilian PD patients and controls. Pearson's correlation coefficient between the test scores was 0.76 (95% CI 0.70-0.81, n = 204, P < 0.001). To calculate reliability measures for each test we used the diagnosis (either PD or control) as outcome variable for separate logistic regression analyses using the score in the UPSIT-40 or the SS-16 as a covariate. The SS-16 specificity was 89.0% with a sensitivity of 81.1% (106 PD and 118 controls). The UPSIT-40 specificity was 83.5% and its sensitivity 82.1% (95 PD and 109 controls). Regression curves were used to associate an individual's smell test score with the probability of belonging to the PD, as opposed to the control group. Our data provide support for the use of the UPSIT-40 and SS-16 to help distinguish early PD from controls.


An overview of potential labor-saving and quality-improving innovations in long-term care for older people.
Increasing demands in long-term care for older people and a decrease in workforce availability can be expected in the future. These developments challenge the sustainability and quality of long-term care for older people. To address these challenges, long-term care organizations are forced to innovate. The aim of this study is to provide an overview of potential labor-saving and quality-improving innovations long-term care organizations are working on and to assess the self-reported extent of effectiveness. This is a descriptive cross-sectional study. In total, 32 long-term care organizations in the region of Limburg in The Netherlands were invited to participate in the study. The inventory was performed by means of semistructured interviews with chief executive officers, managers, and staff members of the long-term care organizations. Based on the interview data, all innovations were described in a standardized form and subsequently checked by the participants. All innovations were clustered into product, process, organizational, and marketing innovations. In total, 26 long-term care organizations delivering home and/or institutional long-term care for older people participated in the study. Overall, 228 innovations were identified; some innovations were described in a similar way by different organizations. The majority of innovations were product innovations (n = 96), followed by organizational innovations (n = 75), and process innovations (n = 42). In addition to the main types, 15 other innovations incorporating characteristics of different types of innovations were detected. Little evidence about the effectiveness of the innovations was reported by the organizations. This study shows that a large number and a broad variety of innovations have been implemented or are currently being developed in long-term care organizations for older people. However, according to the organizations, there is relatively little (scientific) evidence confirming the effectiveness of these innovations. More research is needed to evaluate the effects of the innovations and to indicate whether they provide real solutions to future challenges.


Circulating tumor cells count predicts survival in colorectal cancer patients.
Data on the potential of circulating tumor cells (CTC) count in predicting overall survival (OS) in patients with colorectal cancer are timely and worthy of interest. This study aimed to evaluate the prognostic role of CTC count in both localized and metastatic colorectal cancer patients. Consecutive patients with histological diagnosis of colorectal cancer were enrolled. CTC count was performed, by using a quantitative immunofluorescence method, at baseline (T0) and 1 month following start of chemotherapy (T1). A CTC count <2 was considered negative, whilst a CTC level >/= 2 was positive. Overall survival was calculated accordingly. A total of 75 colorectal cancer patients were enrolled, including 54 stages I-III and 21 stage IV patients. Overall, 21 (28%) patients had a positive CTC count at baseline, and it was significantly associated with a worse prognosis as compared to a negative status (OS: 36.2 vs. 61.6 months; P = 0.002). CTC count remained positive after chemotherapy in 22.4% of the patients and it was an independent prognostic factor of OS (P = 0.03; Hazard Ratio: 3.55; 95% CI: 1.1-11.5). This study found that the presence of CTCs is associated with a reduced survival in colorectal cancer patients. Further studies aimed at testing such a predictive value in early stage colorectal cancer are awaited.


Regulatory domains of erythrocyte ankyrin.
This report provides evidence for regulatory domains of erythrocyte ankyrin that modulate associations of this protein with the anion transporter and spectrin. Two domains have been identified that are located at opposite ends of the polypeptide chain. One domain (Mr = 20,000), which is released by calpain, is primarily involved in regulation of the association of ankyrin with the anion transporter. The Mr = 195,000 fragment remaining after calpain cleavage binds to ankyrin-depleted inside-out vesicles with a 8-fold reaction in affinity, although with a 2-fold increase in number of high affinity sites. Cleavage of ankyrin by calpain induces a reduction in the frictional ratio from 1.55 to 1.33 suggesting either that the calpain-sensitive domain is present as a tail extending from a globular domain, or that upon cleavage ankyrin undergoes a major change in conformation. The other proposed regulatory domain is missing in protein 2.2, a form of ankyrin present in human erythrocytes that has a molecular weight about 29,000 smaller than ankyrin. Protein 2.2 is distinct from the calpain fragment based on peptide maps and reaction with domain-specific antibodies. The activity of the domain deleted from protein 2.2 has been inferred by comparison of ankyrin and protein 2.2, with the assumption that differences between these proteins are due to the missing domain. Protein 2.2 is an activated form of ankyrin that has a 3-fold higher affinity for spectrin and binds to twice the number of high affinity anion transporter sites. These observations suggested that removal of terminal domains of ankyrin may have a physiological role in modulation of ankyrin activity.


Variance and covariance components for liability of piglet survival during different periods.
Variance and covariance components for piglet survival in different periods were estimated from individual records of 133 004 Danish Landrace piglets and 89 928 Danish Yorkshire piglets, using a liability threshold model including both direct and maternal additive genetic effects. At the individual piglet level, the estimates of direct heritability in Landrace were 0.035, 0.057 and 0.027, and in Yorkshire the estimates were 0.012, 0.030 and 0.025 for liability of survival at farrowing (SVB), from birth to day 5 (SV5) and from day 6 to weaning (SVW), respectively. The estimates of maternal heritability for SVB, SV5 and SVW were, respectively, 0.057, 0.040 and 0.030 in Landrace, and 0.050, 0.038 and 0.019 in Yorkshire. Both direct and maternal genetic correlations between the three survival traits were low and not significantly different from zero, except for a moderate direct genetic correlation between SVB and SV5 and between SV5 and SVW in Landrace. Direct and maternal genetic correlations between piglet birth weight (BW) and SV5 were moderately high, but the correlations between BW and SVB and between BW and SVW were low and most of them were not significantly different from zero. These results suggest that effective genetic improvement in piglet survival before weaning by selection should be based on both direct and maternal additive genetic effects and treat survival in different periods as different traits.


Patient data acquisition.
Patient data are acquired in three ways: by direct interrogation; physiological measurements; and analysis of specimens, signals, and images. When computers are used to acquire information from the patient directly, difficulties arise from the lack of standardized patient medical history, the complexities of natural language processing, and the problems of man/machine communication (patient with computer terminal, and physician with computer-generated history). A great variety of data input devices have been used for the acquisition of the patient medical history. Most have been extensively used in multiphasic health testing programs, and this experience is freely drawn upon in this paper.


Computational mechanistic elucidation of the rare earth metal-mediated cycloamidination of aminoalkenes with nitriles.
A detailed computational probe of the rare earth metal-mediated intramolecular amidination of aminoalkenes with nitriles by an archetypical [Y{N(SiMe3)2}3] precatalyst is presented. The mechanistic picture derived from smooth energy profiles, acquired by employing a reliable computational protocol applied to a realistic catalyst model, conforms to available experimental data that includes a significant primary KIE. Sequential Y-N silylamide aminolysis transforms the precatalyst into a multitude of silylamide/amide compounds, of which the bis-amine coordinated [Y{N(SiMe3)2}(NRR')2] is the most abundant, capable of promoting cycloamidination. Nitrile insertion is irreversible and readily furnishes the κ2-N-amidinate yttrium intermediate, which readily rearranges into more stable isomers featuring κ2-N,Δ and κ1-N(imine) amidinate ligations. Its bis-amine adduct likely represents the catalyst resting state. The alkenylamidine becomes accessible through kinetically affordable Y-N amidinate bond protonolysis, which can best be viewed as a kinetically mobile equilibrium that favours the amidinate. The generation of 2-imidazoline product via N-C bond forming amidinate cyclisation favours a stepwise σ-insertive cyclisation/Y-C alkyl aminolysis sequence over an otherwise kinetically prohibitive proton-triggered concerted N-C/C-H bond forming process. The operative σ-insertive pathway entails reversible olefin 1,2-insertion followed by turnover-limiting Y-C alkyl aminolysis at the short-lived 4-imidazolylalkyl intermediate. The DFT estimated primary KIE associated with aminolysis is gratifyingly close to the observed value, thereby supporting the derived mechanistic view.


Comprehensive evaluation of finger flexor tendon entheseal soft tissue and bone changes by ultrasound can differentiate psoriatic arthritis and rheumatoid arthritis.
To determine whether a detailed sonographic evaluation of the hand flexor tendon compartment could help differentiate between psoriatic arthritis (PsA) and rheumatoid arthritis (RA). Thirty-seven patients with PsA, 47 with RA and 10 healthy controls (HC) had flexor tendon (FT) compartment imaging of the dominant hand 2nd to 4th tendons using grey scale (GS) and power Doppler (PD) ultrasound (US) with evaluation for tenosynovitis, peri-tendinous lesions, soft tissue oedema and bony changes at FT insertions. 24/37 PsA and 19/47 RA cases had morning stiffness and 19/37 PsA and 10/47 RA had swollen and/or tender fingers. Tenosynovitis was more common in PsA (25/37) despite higher DAS28 scores in RA (25/37 versus 10/45; p<0.001). Peri-tendinous dermal soft tissue oedema with associated PD signal was evident in one third of PsA patients but in no RA patients (p=0.003). Flexor tendon enthesopathy including new bone formation at the insertional site was significantly more common in PsA (p=0.001). Considering a total inflammatory score per patient summing up the three modifications of the flexor tendon (tenosynovitis, peri-tendinous oedema and insertional enthesophytes) the difference between PsA and RA remained statistically significant (p<0.001). Our study adds to the growing body of literature that high resolution US of the hand FT compartment may help differentiate between RA and PsA, which needs assessment in the diagnostic setting.


Genome-wide methylation patterns in Salmonella enterica Subsp. enterica Serovars.
The methylation of DNA bases plays an important role in numerous biological processes including development, gene expression, and DNA replication. Salmonella is an important foodborne pathogen, and methylation in Salmonella is implicated in virulence. Using single molecule real-time (SMRT) DNA-sequencing, we sequenced and assembled the complete genomes of eleven Salmonella enterica isolates from nine different serovars, and analysed the whole-genome methylation patterns of each genome. We describe 16 distinct N6-methyladenine (m6A) methylated motifs, one N4-methylcytosine (m4C) motif, and one combined m6A-m4C motif. Eight of these motifs are novel, i.e., they have not been previously described. We also identified the methyltransferases (MTases) associated with 13 of the motifs. Some motifs are conserved across all Salmonella serovars tested, while others were found only in a subset of serovars. Eight of the nine serovars contained a unique methylated motif that was not found in any other serovar (most of these motifs were part of Type I restriction modification systems), indicating the high diversity of methylation patterns present in Salmonella.


Schizophrenia is defined by cell-specific neuropathology and multiple neurodevelopmental mechanisms in patient-derived cerebral organoids.
Due to an inability to ethically access developing human brain tissue as well as identify prospective cases, early-arising neurodevelopmental and cell-specific signatures of Schizophrenia (Scz) have remained unknown and thus undefined. To overcome these challenges, we utilized patient-derived induced pluripotent stem cells (iPSCs) to generate 3D cerebral organoids to model neuropathology of Scz during this critical period. We discovered that Scz organoids exhibited ventricular neuropathology resulting in altered progenitor survival and disrupted neurogenesis. This ultimately yielded fewer neurons within developing cortical fields of Scz organoids. Single-cell sequencing revealed that Scz progenitors were specifically depleted of neuronal programming factors leading to a remodeling of cell-lineages, altered differentiation trajectories, and distorted cortical cell-type diversity. While Scz organoids were similar in their macromolecular diversity to organoids generated from healthy controls (Ctrls), four GWAS factors (PTN, COMT, PLCL1, and PODXL) and peptide fragments belonging to the POU-domain transcription factor family (e.g., POU3F2/BRN2) were altered. This revealed that Scz organoids principally differed not in their proteomic diversity, but specifically in their total quantity of disease and neurodevelopmental factors at the molecular level. Single-cell sequencing subsequently identified cell-type specific alterations in neuronal programming factors as well as a developmental switch in neurotrophic growth factor expression, indicating that Scz neuropathology can be encoded on a cell-type-by-cell-type basis. Furthermore, single-cell sequencing also specifically replicated the depletion of BRN2 (POU3F2) and PTN in both Scz progenitors and neurons. Subsequently, in two mechanistic rescue experiments we identified that the transcription factor BRN2 and growth factor PTN operate as mechanistic substrates of neurogenesis and cellular survival, respectively, in Scz organoids. Collectively, our work suggests that multiple mechanisms of Scz exist in patient-derived organoids, and that these disparate mechanisms converge upon primordial brain developmental pathways such as neuronal differentiation, survival, and growth factor support, which may amalgamate to elevate intrinsic risk of Scz.


Semilogarithmic Plots of Data Which Reflect a Continuum of Exponential Processes.
A set of experimental data which describes a curve on semilogarithmic paper may reflect the operation of a continuum of exponential processes, particularly in biological systems where variability is expected. The "backward projection" technique for graphic separation of exponential processes is most misleading when applied to systems in which the processes are distributed so that the standard deviation of halftimes is large compared to the mean halftime.


Beneficial effect of chronic Staphylococcus aureus infection in a model of multiple sclerosis is mediated through the secretion of extracellular adherence protein.
Bacterial infections have been assumed to worsen multiple sclerosis (MS) disease symptoms and to lead to increased neurodegeneration. However, the underlying biological mechanisms for these effects are complex and poorly understood. Here, we assessed the disease-modulating effects of chronic infection with Staphylococcus aureus, a common human pathogen, on the clinical course and the extent of neurodegeneration in experimental autoimmune encephalomyelitis (EAE), an animal model of MS. To conduct this study, we established a persistent chronic infection in female brown Norway rats by inoculating Staphylococcus aureus (S. aureus) bacteria in a subcutaneously implanted tissue cages. In this study, we observed that the introduction of a localized S. aureus infection during the subclinical phase of EAE induced a chronic systemic inflammatory response, consisting of increased T- and B-cell counts and systemic production of proinflammatory cytokines. Unexpectedly, the S. aureus infection completely prevented the development of clinical EAE, and markedly reduced inflammatory infiltration and demyelination of the optic nerve, while it increased the number of surviving retinal neurons. Using a S. aureus strain that lacked the extracellular adherence protein (Eap), we determined that the extracellular adherence protein is at least partially responsible for the inhibitory effect of S. aureus infection on autoimmune inflammation of the central nervous system. Our results demonstrate for the first time that chronic infection with S. aureus has a beneficial effect on EAE, indicating a dual role of infection in the pathogenesis of MS. We also showed that secretion of Eap by S. aureus plays a major role in preventing autoimmune inflammation of the CNS. Moreover, we identified Eap as a factor responsible for this protective effect.


Approaching ultimate resolution for soft x-ray spectrometers.
We explore the potential performance of soft x-ray spectrometers based on the use of varied-line-spacing spherical diffraction gratings (VLS-SG). The quantitative assessment is based on an optimization procedure to obtain both negligible optical aberrations at full illumination of the grating and a quasi linear focal curve. It involves high-order optical aberration cancellation to calculate the focal curves. We also examine the validity of small divergence closed-form formulas describing the light path function. Optimizing the optical and geometric parameters gives an ultimate resolving power, at 930 eV, of between 10,800 for a 3 m long instrument and 34,000 for an 11 m spectrometer according to the Rayleigh criterion. Typical fabrication tolerances would scale these values down by about 10%. The findings are validated by ray-tracing simulations.


The course of posttraumatic stress disorder in a follow-up study of survivors of the Oklahoma City bombing.
The course of posttraumatic stress disorder (PTSD) in populations directly exposed to terrorist attacks is of major importance in the post-9/11 era. Because no systematic diagnostic studies of the most highly exposed individuals of the 9/11 terrorist attacks have yet been done, the Oklahoma City bombing remains a unique opportunity to examine PTSD over time in high-exposure terrorist victims. This study assessed 137 survivors in the direct path of the explosion at approximately 6 and 17 months postdisaster, using the Diagnostic Interview Schedule. Combined index and follow-up data yielded a higher (41%) incidence of PTSD than detected at index (32%) or follow-up (31%). All PTSD was chronic (89% unremitted at 17 months) with no delayed-onset cases. The avoidance and numbing symptom group C, unlike groups B and D alone, was pivotal to current PTSD status and was associated with indicators of functioning at index and follow-up. The findings at index were sustainable. This follow-up study confirmed the immediacy of onset of PTSD and its persistence over time, pointing to the need for early interventions that continue over the long term. Group C avoidance and numbing symptoms may aid in early recognition of PTSD and in predicting long-term functioning.


Treadmill vs. overground running gait during childhood: a qualitative and quantitative analysis.
Conventional gait labs are limited in their ability to study running gait due to their size. There is no consensus in the literature regarding the ability to extrapolate results for adult treadmill running to overground. This comparison has not been studied in children. Twenty-four healthy children (mean age 11.7) ran overground at a slow running speed while motion capture, ground reaction force, and surface electromyography (EMG) data were obtained. The same data were then collected while participants ran for 6min on an instrumented treadmill at a speed similar to their overground speed. The kinematic, kinetic, and EMG data for overground and treadmill running were compared. Sagittal plane kinematics demonstrated similar hip and knee waveforms with the exception of more knee extension just before toe off. Ankle kinematic waveforms were similar during stance phase but treadmill running demonstrated decreased dorsiflexion during swing. Kinetic data was significantly different between the two conditions with treadmill running having a more anterior ground reaction force compared to overground. Due to the numerous differences between overground and treadmill gait demonstrated in this study, it is felt that the use of an instrumented treadmill is not a surrogate to the study of overground running in a pediatric population. This data set will function as a normative data set against which future treadmill studies can be compared.


Emotional and behavioural resilience to multiple risk exposure in early life: the role of parenting.
Ecological and transactional theories link child outcomes to neighbourhood disadvantage, family poverty and adverse life events. Traditionally, these three types of risk factors have been examined independently of one another or combined into one cumulative risk index. The first approach results in poor prediction of child outcomes, and the second is not well rooted in ecological theory as it does not consider that distal risk factors (such as poverty) may indirectly impact children through proximal risk factors (such as adverse life events). In this study, we modelled simultaneously the longitudinal effects of these three risk factors on children's internalising and externalising problems, exploring the role of parenting in moderating these effects. Our sample followed 16,916 children (at ages 3, 5 and 7 years; N = 16,916; 49% girls) from the UK Millennium Cohort Study. Parenting was characterised by quality of parent-child relationship, parental involvement in learning and parental discipline. Neighbourhood disadvantage, family poverty and adverse events were all simultaneously related to the trajectories of both outcomes. As expected, parenting moderated risk effects. Positive parent-child relationship, rather than greater involvement or authoritative discipline, most consistently 'buffered' risk effects. These findings suggest that a good parent-child relationship may promote young children's emotional and behavioural resilience to different types of environmental risk.


High-Reliability Principles Must Be Tied to Value-Based Outcomes.
WellStar Health System and the Medical University of South Carolina (MUSC), highlighted in this issue's feature articles, are two organizations seeking to drive high reliability by educating leaders and incorporating high-reliability principles into their quality improvement (QI) efforts. The organizations have taken slightly different approaches to executing on high reliability, yet both are encouraged by the apparent success of high-reliability principles in other industries.The high-reliability framework is often applied to healthcare despite the limitations of comparing healthcare organizations to nuclear reactors, commercial airlines, and aircraft carriers. Notably, these industries were classified as "highly reliable" after the fact, meaning their employees and leadership already had existing routines and qualities that researchers would describe, after direct observation, as a shared distinguishing feature. Thus, in contrast to Lean, Six Sigma, and other QI movements that have been applied in healthcare, industries such as nuclear power and aviation came with tools and quantitative processes already embedded. The high-reliability framework is qualitative, while the actual hard, quantitative tools and processes differ by industry. This fact makes adopting high-reliability principles difficult because the tools must be created and scaled for each industry. Healthcare is still in the early stages of building these tools to support the high-reliability framework, and the articles by Saunders and Brennan (at WellStar) and Cawley and Scheurer (at MUSC) describe early attempts to provide insights into launching high-reliability principles and tools in healthcare.High reliability is a theoretical construct that is difficult to implement without a concrete framework for execution. The five characteristics that frame a high-reliability organization (HRO), as outlined by Cawley and Scheurer (citing ), are (1) preoccupation with failure, (2) reluctance to simplify interpretations, (3) sensitivity to operations, (4) commitment to resilience, and (5) deference to expertise. Although these elements point in the right direction, they remain theoretical if not tethered to an organization's operating cycle.


Activation of inducible nitric oxide synthase results in nitric oxide-mediated radiosensitization of hypoxic EMT-6 tumor cells.
EMT-6 cells treated for 16 h with 1-10 units/ml IFN-gamma showed a gradual activation of inducible nitric oxide synthase (iNOS) in Western and Northern blots, a simultaneous raise in NO output, and an increase in hypoxic cell radiosensitivity almost to the level of aerobic cells. Both the NO signal and radiosensitization were counteracted by the NO scavenger oxyhemoglobin, by the specific iNOS inhibitor aminoguanidine, and by the L-arginine analogue N(G)-monomethyl-L-arginine. Collectively, these data demonstrate that IFN-gamma can radiosensitize EMT-6 cells through iNOS induction and that NO is the effector molecule responsible for radiosensitization. Compared with the spontaneous NO releaser (2)-1-[N-(3-ammoniopropyl)-N-(n-propyl)amino)diazen-1-ium -1,2-diolate], the iNOS-generated NO signal appeared to be 10 times lower yet resulting in the same enhancement ratio of 2.4. Direct stimulation of NO synthesis in tumor cells through the L-arginine/iNOS pathway represents a novel approach to exploit the radiosensitizing properties of NO.


Multifunctional 6-fluoro-3-[3-(pyrrolidin-1-yl)propyl]-1,2-benzoxazoles targeting behavioral and psychological symptoms of dementia (BPSD).
Patients suffering from dementia experience cognitive deficits and 90% of them show non-cognitive behavioral and psychological symptoms of dementia (BPSD). The spectrum of BPSD includes agitation, depression, anxiety and psychosis. Antipsychotics, e.g. quetiapine, have been commonly used off-label to control the burdensome symptoms, though they cause serious side effects and further cognitive impairment. Therefore, the development of targeted therapy for BPSD, suitable for elderly patients, remains relevant. A multitarget-directed ligand, acting on serotonin 5-HT2A and dopamine D2 receptors (R) and thus exerting anti-aggressive and antipsychotic activity, as well as on 5-HT6Rs and 5-HT7Rs (potential pro-cognitive, antidepressant and anxiolytic activity), poses a promising strategy for the treatment of BPSD. Antitargeting muscarinic M3R and hERG channel is expected to reduce the risk of side effects. We obtained a series of stereoisomeric compounds by combining 6-fluoro-1,2-benzoxazole moiety and arylsulfonamide fragment through pyrrolidin-1-yl-propyl linker. N-[(3R)-1-[3-(6-fluoro-1,2-benzoxazol-3-yl)propyl]pyrrolidin-3-yl]-1-benzothiophene-2-sulfonamide showed a substantial affinity for the targets of interest (pKi = 8.32-9.35) and no significant interaction with the antitargets. Functional studies revealed its antagonist efficacy (pKB = 7.41-9.03). The lead compound showed a promising profile of antipsychotic-like activity in amphetamine- and MK-801-induced hyperlocomotion (MED = 2.5 mg/kg), antidepressant-like, as well as anxiolytic-like activity in mice (MED = 0.312 and 1.25 mg/kg in the forced swim and four-plate tests, respectively). Notably, the novel compound didn't affect spontaneous locomotor activity, nor induced catalepsy or memory deficits (step-through passive avoidance test) in therapeutically relevant doses, which proved its benign safety profile. The overall pharmacological characteristics of the lead compound outperformed the reference drug quetiapine, making it a promising option for evaluation in the treatment of BPSD.


Peony glycosides protect against corticosterone-induced neurotoxicity in PC12 cells.
Preclinical and clinical investigations have shown the involvement of dysregulation of hypothalamic-pituitary-adrenal (HPA) axis in the pathogenesis of depression. Hypercortisolemia and the associated hippocampal atrophy were observed in patients with depression, which could be ameliorated by the treatment with antidepressants. Therefore, neuroprotection has been proposed to be one of the acting mechanisms of antidepressant. Previous studies in our laboratory have demonstrated the antidepressant-like activity of total glycosides of peony (TGP) in mice. This study aimed to examine the effect of TGP treatment on corticosterone-induced neurotoxicity in cultured rat pheochromocytoma (PC12) cells. Treating the cells with corticosterone at 200 muM for 48 h caused apoptotic cell death. The cytotoxicity was associated with the activation of caspase-3 activity and the decrease in the mRNA ratio of bcl-2 to bax. TPG treatment at increasing doses (1-10 mg/l) protected against the corticosterone-induced toxicity in PC12 cells in a dose-dependent manner. The cytoprotection afforded by TGP treatment was associated with the inhibition of caspase-3 activity and the up-regulation of bcl-2/bax mRNA ratio. The anti-apoptotic effect of TGP is therefore likely mediated by the suppression of the mitochondrial pathway leading to apoptosis.


A rare case of iron-pill induced gastritis in a female teenager: A case report and a review of the literature.
The treatment of iron-deficiency anemia with oral iron supplements can present side-effects on the GI tract mucosa including necrosis, ulceration, or ischemia. The particular endoscopic findings and the histopathological exam will establish the diagnosis of erosive gastritis with iron deposits in the gastric mucosa. We present the case of a 14-year-old female admitted in our clinic for upper digestive hemorrhage, nausea, melena, and abdominal pain. Her personal history revealed iron deficiency anemia receiving oral iron supplements for approximately 2 weeks. The laboratory tests at the moment of admission pointed out anemia, increased level of serum iron, increased liver transaminases, a decreased level of ferritin, but with normal levels of both total iron-binding capacity and transferrin. The eso-gastro-duodenoscopy revealed multiple brown deposits on the surface of the gastric mucosa and multiple hemorrhagic lesions, under the aspect of erosions all over the gastric mucosa, but more severe in the antral part, and the histopathological exam confirmed the presence of iron deposits at this level. Iron-pill induced gastritis is a rare, under-diagnosed entity that can be present even at pediatric ages with potential severe clinical impact.


A Brief Review of Personality in Marathon Runners: The Role of Sex, Age and Performance Level.
The participation of recreational runners in sport events ranging from 5 km to ultra-endurance races have increased dramatically during the last decades and this phenomenon has attracted scientific interest. Most research has focused on the physiological characteristics of these runners and less in their psychological characteristics. Therefore, the aim of the present study was to review the existing knowledge with regards to the personality of recreational endurance runners and the role of sex, age and performance. It was concluded that limited information was available with regards to the personality of recreational marathon runners. So far, our knowledge on the personality of marathon runners relied on studies conducted a few decades ago, mostly on competitive marathon runners, highlighting the need for original research on recreational runners.


Influence of renal failure on the disposition of morphine, morphine-3-glucuronide and morphine-6-glucuronide in sheep during intravenous infusion with morphine.
The influence of experimentally induced renal failure on the disposition of morphine, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) was examined in seven sheep infused intravenously with morphine for 6 hr. Between 5 and 6 hr, blood was collected from the aorta, pulmonary artery, hepatic, hepatic portal and renal veins and posterior vena cava. Additional samples from the aorta and urine were collected up to 144 hr. Morphine, M3G and M6G were determined in plasma and urine by high-performance liquid chromatography. Constant concentrations of morphine, but not of M3G and M6G, were achieved in plasma between 5 and 6 hr. Significant (P < .001) extraction of morphine by the liver (0.72 +/- 0.05) and kidney (0.42 +/- 0.15) occurred. Compared with sheep with normal kidneys (Milne et al., 1995), renal failure did not alter (P = .11) the mean total clearance of morphine (1.5 +/- 0.3 liters/min); clearance by the kidney was less (P < .001). However, a paired comparison using sheep common to this study and from the study when their kidneys were normal revealed a significant reduction in mean total clearance of 25%. The renal extraction of M3G and M6G and urinary recovery of the dose as summed morphine, M3G and M6G were reduced by renal failure. The kidney metabolized morphine to M3G. The data suggest that nonrenal elimination of M3G becomes more important during renal failure.


Polynuclear aromatic hydrocarbons in an agricultural soil: long-term changes in profile distribution.
Soil profile samples collected from the same plot at Rothamsted Experimental Station in southeast England in 1893, 1944 and 1987 have been analysed for polynuclear aromatic hydrocarbons (PAHs). The total PAH burden of the plough layer (0-23 cm) has increased approximately four- to five-fold since the 1890s, with some compounds (notably benzo(b)fluoranthene, benzo(a)pyrene and pyrene) showing substantially greater increases. Average rates of increase in the plough layer for individual PAHs in the Rothamsted plots over the century since c. 1890 vary between 0.01 and 0.67 mg m(-2) year(-1). It is concluded that atmospheric deposition from natural sources has been augmented in recent years by regional fallout of anthropogenically-generated PAHs derived from the combustion of fossil fuels. The total PAH content of the 1893 Rothamsted samples was similar to that observed in soils from contemporary isolated/rural locations in the UK and showed little surface enrichment. By 1987 the surface soil at Rothamsted had been enriched in all PAH compounds measured by a factor of between 1.3 (acenaphthalene) and over 20 (benzo(a)pyrene). Increases in the PAH content of the 23-46 cm subsurface layer indicate migration of PAHs from the plough layer. Net average annual migration rates ranged from 0.01-0.14 mg m(-2) year(-1) for individual PAHs, and the rate appeared to be primarily a function of the plough layer PAH content, rather than physical/chemical properties of the individual compounds. This suggests particle-bound translocation as the dominant mechanism of PAH migration.


Incompatibility groups and the classification of fi - resistance factors.
Incompatibility between R factors has been reported by several authors, and four incompatibility groups have already been described by Datta and Hedges among Rfi(-) factors. The stability of 12 plasmids in pairs was studied after 116 crosses, and five new groups were found, designated 5, 6, 7, 8, and 9. Each plasmid studied belongs to one single group. Incompatibility between plasmids in pairs is a clear-cut phenomenon, is easy to observe, and can provide a reliable method for recognizing and classifying resistance factors, and for tracing their spread among bacterial species.


Coordinated regulation of mitochondrial topoisomerase IB with mitochondrial nuclear encoded genes and MYC.
Mitochondrial DNA (mtDNA) is entirely dependent on nuclear genes for its transcription and replication. One of these genes is TOP1MT, which encodes the mitochondrial DNA topoisomerase IB, involved in mtDNA relaxation. To elucidate TOP1MT regulation, we performed genome-wide profiling across the 60-cell line panel (the NCI-60) of the National Cancer Institute Developmental Therapeutics Program. We show that TOP1MT mRNA expression varies widely across these cell lines with the highest levels in leukemia (HL-60, K-562) and melanoma (SK-MEL-28), intermediate levels in breast (MDA-MB-231), ovarian (OVCAR) and colon (HCT-116, HCT-15, KM-12), and lowest levels in renal (ACHN, A498), prostate (PC-3, DU-145) and central nervous system cell lines (SF-539, SF-268, SF-295). Genome-wide analyses show that TOP1MT expression is significantly correlated with the other mitochondrial nuclear-encoded genes including the mitochondrial nucleoid genes, and demonstrate an overall co-regulation of the mitochondrial nuclear-encoded genes. We also find very high correlation between the expression of TOP1MT and the proto-oncogene MYC (c-myc). TOP1MT contains E-boxes (c-myc binding sites) and TOP1MT transcription follows MYC up- and down-regulation by MYC promoter activation and siRNA against MYC. Our finding implicates MYC as a novel regulator of TOP1MT and confirms its role as a master regulator of MNEGs and mitochondrial nucleoids.


Stability of single dispersed silver nanoparticles in natural and synthetic freshwaters: Effects of dissolved oxygen.
Silver nanoparticles (AgNPs) are increasingly used in various commercial products. This increased use raises ecological concerns because of the large release of AgNPs into the environment. Once released, the local water chemistry has the potential to influence the environmental fates and behaviors of AgNPs. The impacts of dissolved oxygen and natural organic matter (NOM) on the dissolution and stability of AgNPs were investigated in synthetic and natural freshwaters for 7 days. In synthetic freshwater, the aggregation of AgNPs occurred due to the compression of the electric double layer, accompanied by the dissolution of AgNPs. However, once oxygen was removed, the highest dissolved Ag (Agdis) concentration decreased from 356.5 μg/L to 272.1 μg/L, the pH of the AgNP suspensions increased from less than 7.6 to more than 8.4, and AgNPs were regenerated by the reduction of released Ag+ by citrate. The addition of NOM mitigated aggregation, inhibited oxidative dissolution and induced the transformation of AgNPs into Ag2S due to the formation of NOM-adsorbed layers, the reduction of Ag+ by NOM, and the high affinity of sulfur-enriched species in NOM for Ag. Likewise, in oxygen-depleted natural freshwaters, the inhibition of oxidative dissolution was obtained in comparison with oxygenated freshwaters, showing a decrease in the maximum Agdis concentration from 137.6 and 57.0 μg/L to 83.3 and 42.4 μg/L from two natural freshwater sites. Our results suggested that aggregation and dissolution of AgNPs in aquatic environments depend on the chemical composition, where oxygen-depleted freshwaters more significantly increase the colloidal stability. In comparison with oxic conditions, anoxic conditions were more favorable to the regeneration of AgNPs by reducing species (e.g., citrate and NOM) and enhanced the stability of nanoparticles. This indicates that some AgNPs will be more stable for long periods in oxygen-deprived freshwaters, and pose more serious environmental risks than that in oxygenated freshwaters.


Real-time PCR for quantitative analysis of human commensal Escherichia coli populations reveals a high frequency of subdominant phylogroups.
Escherichia coli is divided into four main phylogenetic groups, which each exhibit ecological specialization. To understand the population structure of E. coli in its primary habitat, we directly assessed the relative proportions of these phylogroups from the stools of 100 healthy human subjects using a new real-time PCR method, which allows a large number of samples to be studied. The detection threshold for our technique was 0.1% of the E. coli population, i.e., 10(5) CFU/g of feces; in other methods based on individual colony analysis, the threshold is 10%. One, two, three, or four phylogenetic groups were simultaneously found in 21%, 48%, 21%, and 8% of the subjects, respectively. Phylogroups present at a threshold of less than 10% of the population were found in 40% of the subjects, revealing high within-individual diversity. Phylogroups A and B2 were detected in 74% and 70% of the subjects, respectively; phylogroups B1 and D were detected in 36% and 32%, respectively. When phylogroup B2 was dominant, it tended not to cooccur with other phylogroups. In contrast, other phylogroups were present when phylogroup A was dominant. These data indicate a complex pattern of interactions between the members of a single species within the human gut and identify a reservoir of clones that are present at a low frequency. The presence of these minor clones could explain the fluctuation in the composition of the E. coli microbiota within single individuals that may be seen over time. They could also constitute reservoirs of virulent and/or resistant strains.


Bilirubin and its oxidative metabolite biopyrrins in patients with acute myocardial infarction.
This study aimed to assess the involvement of bilirubin and its oxidative metabolite biopyrrin in patients with acute myocardial infarction (AMI) and to determine the responsible organs that overproduce these molecules. One hundred thirteen consecutive patients hospitalized for AMI were analyzed. Levels of serum bilirubin, plasma and urinary biopyrrins were measured on the day of admission, day 2, 3, 7 and 14. Expressions of biopyrrins and heme oxygenase-1 (HO-1), a stress-responsive bilirubin-producing enzyme, in heart, aorta, kidney, liver and lung were immunostained with autopsied specimens. Serum bilirubin, plasma and urinary biopyrrins were increased within 24 hr, formed a peak on day 3 and then decreased by day 14. These three parameters were well correlated to each other. The maximum biopyrrin elevation was higher in death cases and associated with impaired left ventricular function. Immunohistochemical analyses revealed biopyrrin accumulation and HO-1 expression in the infracted myocardium. Immunoreactive HO-1 and biopyrrins were also observed in renal tubular cells, aortic wall and lung. Serum bilirubin and its metabolite biopyrrins were elevated in patients with AMI. Plasma and urinary biopyrrin elevation were associated with mortality and morbidity. Induction of anti-oxidative enzyme HO-1 seemed to be involved in the activation of bilirubin/biopyrrin pathway.


Is Nontarget Analysis Ready for Regulatory Application? Influence of Peak-Picking Algorithms on Data Analysis.
The use of peak-picking algorithms is an essential step in all nontarget analysis (NTA) workflows. However, algorithm choice may influence reliability and reproducibility of results. Using a real-world data set, the aim of this study was to investigate how different peak-picking algorithms influence NTA results when exploring temporal and/or spatial trends. For this, drinking water catchment monitoring data, using passive samplers collected twice per year across Southeast Queensland, Australia (n = 18 sites) between 2014 and 2019, was investigated. Data were acquired using liquid chromatography coupled to high-resolution mass spectrometry. Peak picking was performed using five different programs/algorithms (SCIEX OS, MSDial, self-adjusting-feature-detection, two algorithms within MarkerView), keeping parameters identical whenever possible. The resulting feature lists revealed low overlap: 7.2% of features were picked by >3 algorithms, while 74% of features were only picked by a single algorithm. Trend evaluation of the data, using principal component analysis, showed significant variability between the approaches, with only one temporal and no spatial trend being identified by all algorithms. Manual evaluation of features of interest (p-value <0.01, log fold change >2) for one sampling site revealed high rates of incorrectly picked peaks (>70%) for three algorithms. Lower rates (<30%) were observed for the other algorithms, but with the caveat of not successfully picking all internal standards used as quality control. The choice is therefore currently between comprehensive and strict peak picking, either resulting in increased noise or missed peaks, respectively. Reproducibility of NTA results remains challenging when applied for regulatory frameworks.


Prolonged life and improved quality for stored skin grafts.
This study is presented as both a follow-up and an extension of previous work on storage techniques for the preservation of skin by refrigeration. Rabbit skin grafts were used to compare the merits of various storage media. One of the media, McCoy's 5A plus serum, gave improved skin graft take and storage half-life. The ability to improve skin graft storage techniques is most beneficial to all aspects of plastic and reconstructive surgery where skin grafting is required. Prolonged graft viability and improved graft quality are of major clinical importance.


Quantitative study of spongiform change in putamen of 24 cases of Creutzfeldt-Jakob disease.
Creutzfeldt-Jakob disease (CJD) is characterized by 4 main neuropathological lesions: spongiform change, neuronal loss, astrocytic gliosis, and accumulation of pathological prion protein (PrPsc), which is partially protease-resistant (PrPres). This study focused on spongiform change (SC) in the putamen. Because SC varies from case to case, we investigated whether its quantification could provide relevant criteria to discriminate types of PrPres in CJD. SC was quantified in 24 CJD cases, 12 with PrPres type 1 (CJD-PrP1) and 12 with PrPres type 2 (CJD-PrP2), compared to 25 control cases. The study was performed by direct microscopy examination (DME) and by semiautomatic quantification (SAQ) using shape and size criteria previously described. These criteria were suitable for SC quantification in putamen in the majority of cases, except for those with microspongiosis. The results obtained by DME and SAQ methods were correlated and SC scores were compared to the types of PrPres. Sporadic CJD cases with PrPres type 2 were more affected by SC than type 1, suggesting that putamen could be a preferential site to distinguish type 1 from type 2 histologically. The origin of the difference in SC intensity according to the type of PrPres is discussed in terms of host and strain factors.


A Maastrichtian insect assemblage from Patagonia sheds light on arthropod diversity previous to the K/Pg event.
Insect faunas from the latest Cretaceous are poorly known worldwide. Particularly, in the Southern Hemisphere, there is a gap regarding insect assemblages in the Campanian-Maastrichtian interval. Here we present an insect assemblage from the Maastrichtian Chorrillo Formation, southern Argentina, represented by well-preserved and non-deformed, chitinous microscopic remains including head capsules, wings and scales. Identified clades include Chironomidae dipterans, Coelolepida lepidopterans, and Ephemeroptera. The assemblage taxonomically resembles those of Cenozoic age, rather than other Mesozoic assemblages, in being composed by diverse chironomids and lepidopterans. To the best of our knowledge, present discovery constitutes the first insect body fossils for the Maastrichtian in the Southern Hemisphere, thus filling the gap between well-known Early Cretaceous entomofaunas and those of Paleogene age. The presented evidence shows that modern clades of chironomids were already dominant and diversified by the end of the Cretaceous, in concert with the parallel radiation of aquatic angiosperms which became dominant in freshwater habitats. This exceptional finding encourages the active search of microscopic remains of fossil arthropods in other geological units, which could provide a unique way of enhancing our knowledge on the past diversity of the clade.


The relationship between objective sleep variables and subjective sleep estimation in schizophrenia.
Few investigations of sleep structure in schizophrenia have concentrated on the relationship between objective and subjective sleep variables. The aim of this study was to assess objective sleep variables and subjective estimation of sleep duration and sleep quality. Polysomnography was performed in 20 chronic patients with schizophrenia during three consecutive nights. After final awakenings subjects answered questions concerning subjective estimations of sleep duration, sleep latency, number of awakenings, and sleep depth. Pearson correlations between ranged subjective reports and objective sleep variables were performed. The results showed a high positive correlation between objective sleep latency and its subjective estimation; a positive correlation between subjective estimation of sleep depth and percentage of slow wave sleep (SWS%); a positive correlation between eye movement (EM) density and subjective estimation of wakefulness during the night; and a negative correlation between EM density and dream reports. We concluded that objective sleep variables are related to subjective sleep estimation in schizophrenic patients. In these patients, EM activity in REM sleep is related to the subjective feeling of wakefulness. (Int J Psych Clin Pract 2000; 4:63-67).


Clinical Course of Infection and Cross-Species Detection of Equine Parvovirus-Hepatitis.
Since its first discovery by Arnold Theiler in 1918, serum hepatitis also known as Theiler's disease has been reported worldwide, causing idiopathic acute hepatitis and liver failure in horses. Recent studies have suggested a novel parvovirus, named equine parvovirus hepatitis (EqPV-H), to be associated with Theiler's disease. Despite the severity and potential fatality of EqPV-H infection, little is known about the possibility of developing chronic infections and putative cross-species infection of equine sister species. In the present longitudinal study, we employed qPCR analysis, serology, and biochemical testing as well as pathology examination of liver biopsies and sequence analysis to investigate potential chronic EqPV-H infection in an isolated study cohort of in total 124 horses from Germany over five years (2013-2018). Importantly, our data suggest that EqPV-H viremia can become chronic in infected horses that do not show biochemical and pathological signs of liver disease. Phylogenetic analysis by maximum likelihood model also confirms high sequence similarity and nucleotide conservation of the multidomain nuclear phosphoprotein NS1 sequences from equine serum samples collected between 2013-2018. Moreover, by examining human, zebra, and donkey sera for the presence of EqPV-H DNA and VP1 capsid protein antibodies, we found evidence for cross-species infection in donkey, but not to human and zebra. In conclusion, this study provides proof for the occurrence of persistent EqPV-H infection in asymptomatic horses and cross-species EqPV-H detection in donkeys.


Mass Disasters and Burnout in Nephrology Personnel: From Earthquakes and Hurricanes to COVID-19 Pandemic.
Mass disasters result in extensive health problems and make health care delivery problematic, as has been the case during the COVID-19 pandemic. Although COVID-19 was initially considered a pulmonary problem, it soon became clear that various other organs were involved. Thus, many care providers, including kidney health personnel, were overwhelmed or developed burnout. This review aims to describe the spectrum of burnout in mass disasters and suggests solutions specifically for nephrology personnel by extending previous experience to the COVID-19 pandemic. Burnout (a psychologic response to work-related stress) is already a frequent part of routine nephrology practice and, not surprisingly, is even more common during mass disasters due to increased workload and specific conditions, in addition to individual factors. Avoiding burnout is essential to prevent psychologic and somatic health problems in personnel as well as malpractice, understaffing, and inadequate health care delivery, all of which increase the health care burden of disasters. Burnout may be prevented by predisaster organizational measures, which include developing an overarching plan and optimizing health care infrastructure, and ad hoc disaster-specific measures that encompass both organizational and individual measures. Organizational measures include increasing safety, decreasing workload and fear of malpractice, optimizing medical staffing and material supplies, motivating personnel, providing mental health support, and enabling flexibility in working circumstances. Individual measures include training on coping with stress and problematic conditions, minimizing the stigma of emotional distress, and maintaining physical health. If these measures fall short, asking for external help is mandatory to avoid an inefficient disaster health care response. Minimizing burnout by applying these measures will improve health care provision, thus saving as many lives as possible.


Pathways From Intimate Partner Violence to Academic Disengagement Among Women University Students.
More knowledge is needed related to collegiate intimate partner violence (IPV) and the pathways between experiencing physical and psychological IPV and academic disengagement. Students in a University System in the southwest completed an online survey including measures of physical and psychological IPV, academic disengagement, sense of community, and safety on campus. Conditional process analyses were used to understand key pathways for 6,818 woman identified students. All models found a significant indirect path between physical and psychological IPV and academic disengagement via depression symptoms. Students' sense of community on campus was associated with less academic disengagement regardless of physical violence. The impact of psychological IPV on disengagement was stronger for those with lower senses of community. Enhancing screening and education, providing effective mental health counseling, and increasing advocacy will help institutions better address IPV.


Establishing performance standards for child development: learnings from the ECDI2030.
Standards of early childhood development (ECD) are needed to determine whether children living in different contexts are developmentally on track. The Early Childhood Development Index 2030 (ECDI2030) is a population-level measure intended to be used in household surveys to collect globally comparable data on one of the indicators chosen to monitor progress toward target 4.2 of the Sustainable Development Goals: The proportion of children aged 24-59 months who are developmentally on track in health, learning and psychosocial well-being. To define performance cut-scores for the ECDI2030 we followed a criterion-referenced standard setting exercise using the modified Angoff method. The exercise gauged the expectations from 15 global experts in ECD and was informed by representative population data collected in Mexico and the State of Palestine. The final calibrated age-specific performance cut-scores were applied to these data to estimate the proportion of children developmentally on track, disaggregated by background characteristics, including the child's sex and attendance to early childhood education. Through a process of standard setting, we generated robust performance standards for the ECDI2030 by establishing five age-specific cut-scores to identify children as developmentally on track. This paper demonstrated how the standard setting methodology, typically applied to measures in the health and education fields, could be applied to a measure of child development. By creating robust criterion-referenced standards, we have been able to ensure that the cut-scores related to age for the ECDI2030 are based on performance standards set by global experts in the ECD field for defining on and off track development.


Expression of HNF-1 alpha and HNF-1 beta in various histological differentiations of hepatocellular carcinoma.
Hepatic nuclear factor 1 (HNF-1) regulates genes in a hepatocyte-specific manner. It has been previously reported that the ratio of HNF-1 alpha and HNF-1 beta mRNA is related to histological differentiation hepatocellular carcinoma (HCC). In this study, the expression levels of the HNF-1 alpha and HNF-1 beta proteins were analysed relatively and quantitatively in various histologically differentiated HCC and surrounding non-cancerous tissues, and HNF-1 alpha binding activity for the AT element of the B domain of the human alpha-fetoprotein enhancer was examined. Western blot analysis demonstrated that HNF-1 alpha protein was expressed at a higher level in well-differentiated HCC tissues than in the surrounding non-HCC tissues; on the other hand, the HNF-1 alpha protein was expressed at lower levels in moderately and poorly differentiated HCCs than in the surrounding non-HCC tissues. The levels of HNF-1 beta expression in well-differentiated and poorly differentiated HCCs were similar to and higher than those found in the respective surrounding non-cancerous portions. In binding assays, HNF-1 binding activity was high in well-differentiated HCC and lower in moderately and poorly differentiated HCCs. Most well-differentiated HCC cases showed immunohistochemical expression of HNF-1 alpha. These findings show that poor histological differentiation of HCC correlates with decreases in the level and activity of HNF-1 alpha proteins.


Hemagglutinins from two influenza virus variants bind to sialic acid derivatives with millimolar dissociation constants: a 500-MHz proton nuclear magnetic resonance study.
The equilibrium binding of influenza virus hemagglutinin to derivatives of its cell-surface ligand, sialic acid, was measured by nuclear magnetic resonance (NMR) spectroscopy. Binding was quantified by observing perturbations of sialic acid resonances in the presence of protein. The major perturbation observed was a chemical shift of the N-acetyl methyl resonance, presumably due to the proximity of the methyl group to tryptophan 153. X-31 hemagglutinin binds to the methyl alpha-glycoside of sialic acid with a dissociation constant of 2.8 mM and does not bind to the methyl beta-glycoside. Replacing the 4-hydroxyl group of sialic acid with an acetyl group has little effect, while replacing the 7-hydroxyl group with an acetyl prevents binding. Experiments with sialylated oligosaccharides confirm literature reports that mutations at amino acid 226 change the specificity of hemagglutinin for alpha(2,6) and alpha(2,3) glycosidic linkages. The NMR line broadening of sialyloligosaccharides suggests that sialic acid is the only component that contacts the protein. Saccharides containing two sialic acid residues appear to have two separate binding modes. Hemagglutinin that has undergone a low pH induced conformational change retains the ability to bind sialic acid.


The importance of habitat resistance for movement decisions in the common lizard, Lacerta vivipara.
Movement behaviour can be influenced by a multitude of biotic and abiotic factors. Here, we investigate the speed of movement in relation to environmental and individual phenotypic properties in subadult common lizards (Lacerta vivipara). We aim to disentangle the importance of substrate, cover, humidity, basking opportunity and individual phenotype on moving tendencies in 12 treatment combinations, at which each lizard was tested. We find that movement behaviour depends on the starting conditions, the physical properties of the dispersal corridor, and on the individuals' phenotype. Specifically, the presence of cover and substrate providing suitable traction in the corridor had positive effects on individual movement decisions. Additionally, we find high phenotypic variation in the propensity to move dependent on the presence of cover. Individual back patterns also strongly affected movement decisions in interaction with the physical properties of the dispersal corridor. Our results highlight the importance of understanding the habitat resistance for movement patterns, with humid habitats with covering vegetation providing the best conditions to initiate movement in the common lizard. In addition, population effects, differences in back pattern phenotype and individual plasticity were identified as key parameters influencing movement behaviour.


The occurrence of brown adipose in man inhabiting the tropics.
Samples of cervical, mediastinal and perirenal fat tissue were obtained from eight subjects (ages: 17 days to 56 years) at necropsy. After formol fixation, the specimens were processed for sectioning and stained with hematoxylin and eosin, periodic acid--Schiff, or Holmes' silver techniques for light microscopy. All the tissue samples showed lobular organisation and a rich vascularity, with argentaffin (putative nerve) fibres at paravascular and parenchymal sites. Three histological forms of brown adipose tissue (BAT) were recognised: homogeneous lobules of multilocular adipocytes (Type I); mixed lobules consisting of islets of multilocular adipocytes surrounded by large unilocular cells (Type II); and lobules consisting mainly of small (less than 40 microns diameter) unilocular adipocytes (Type III). In two subjects (ages 17 days and 52 years), BAT (Types I and II respectively) was observed in tissue samples from all three sites. Perirenal samples from three other donors (ages 26-35 years) showed the presence of Type II or III BAT. Mediastinal BAT (Type II) was found in two subjects (26- and 35-years old). The significance of BAT in man inhabiting a warm environment is discussed.