Ethanol effects on delayed spatial matching as modeled by a negative exponential forgetting function.
Performance on delayed matching-to-position as a function of ethanol was investigated in rats and dose effects assessed by fitting an exponential decay model of forgetting to signal detection sensitivity scores. Three ethanol doses (0.25, 0.50, and 0.75 g/kg) and one isovolume saline control were examined. For further comparison, one dose of chlordiazepoxide (CDP; 5 mg/kg) and its saline control were also given. Forgetting functions were reasonably well described by the decay model under all treatment conditions. In addition, the functions's decay constant (-b) proved to be differentially sensitive to both drug and dose effects. Reduced decay estimates were obtained following the two lowest ethanol doses, the reduction being statistically significant for the 0.25 g/kg dose. In contrast, the function estimate of initial sensitivity, the intercept parameter (SI0), was not significantly affected by ethanol. Consistent with the low-dose ethanol effects, CDP significantly decreased the value of the decay parameter while leaving the intercept parameter unaffected. But unlike ethanol, the variance accounted for by the model for the individual data was less following CDP administration. Drug effects were interpreted using the exponential decay model of forgetting, and the results suggest independent discriminative control over SI0 and b. The significant effect of the low-dose sedative-hypnotics upon b, with no attendant effects upon SI0, is suggested to result from enhanced, spontaneous, delay interval mediation.


Developmental exposure to lead and late life abnormalities of nervous system.
Role of developmental exposure to environmental agents in altering the disease process is well known. Exposure to chemical agents at critical periods of development may cause some permanent changes in the functioning of various vital systems including the nervous system in the organisms. It is not surprising to see an extensive response due to exposure to chemical agents early in life as the organ systems are more vulnerable to chemical insults during developmental stages. In some cases the response to low level environmental insults may not be obvious until adult or old age. Results from several studies have shown such latency in response to the nervous system leading to neurodegeneration in old age. Studies conducted in murine and primate models provided ample evidence for the association of developmental exposure to low levels of heavy metal lead (Pb) and Alzheimer's disease-like pathology during senescence. It is not clear about the reasons behind such response; however, the contribution of epigenetic mechanisms could explain the role of early events in life in inducing the late life abnormalities of nervous system. It is possible that environmental agents epigenetically modulate the gene regulation to persist the response silent for a long period of time and to result pathological outcomes significantly later in life. This article will summarize the association of early life exposure to environmental agents and late-life abnormalities with an emphasis on developmental exposure to Pb and neurodegeneration in old age.


Assessing the generalizability of prognostic information.
Physicians are often asked to make prognostic assessments but often worry that their assessments will prove inaccurate. Prognostic systems were developed to enhance the accuracy of such assessments. This paper describes an approach for evaluating prognostic systems based on the accuracy (calibration and discrimination) and generalizability (reproducibility and transportability) of the system's predictions. Reproducibility is the ability to produce accurate predictions among patients not included in the development of the system but from the same population. Transportability is the ability to produce accurate predictions among patients drawn from a different but plausibly related population. On the basis of the observation that the generalizability of a prognostic system is commonly limited to a single historical period, geographic location, methodologic approach, disease spectrum, or follow-up interval, we describe a working hierarchy of the cumulative generalizability of prognostic systems. This approach is illustrated in a structured review of the Dukes and Jass staging systems for colon and rectal cancer and applied to a young man with colon cancer. Because it treats the development of the system as a "black box" and evaluates only the performance of the predictions, the approach can be applied to any system that generates predicted probabilities. Although the Dukes and Jass staging systems are discrete, the approach can also be applied to systems that generate continuous predictions and, with some modification, to systems that predict over multiple time periods. Like any scientific hypothesis, the generalizability of a prognostic system is established by being tested and being found accurate across increasingly diverse settings. The more numerous and diverse the settings in which the system is tested and found accurate, the more likely it will generalize to an untested setting.


Inotropic effect of digoxin in humans: mechanistic pharmacokinetic/pharmacodynamic model based on slow receptor binding.
The purpose of this study was to construct a mechanistic pharmacokinetic/pharmacodynamic (PK/PD) model for digoxin that describes the relationship between plasma concentration and inotropic response. On the basis of results obtained in the isolated perfused rat heart, a PK/PD model for digoxin in humans was developed. In fitting the model to previously published bolus dose and concentration clamp data (shortening of electromechanical systole), the plasma concentration-time curves were used as forcing functions in the computer program ADAPT II. The mechanistic approach allowed a modeling of digoxin pharmacodynamics which is consistent with available inotropic response data. The estimates of the receptor binding parameters were in the same order of magnitude as those measured in vitro for ouabain. The mechanistic model explained the parameters of the empirical link model (EC50, Emax and delay time tau) in terms of the underlying processes, suggesting that the long equilibration half-time of 13 h is due to slow receptor binding. The empirical link model, in contrast, is not compatible with a noninstantaneous receptor binding process and led to estimates of the delay time tau that were dependent on the digoxin administration schedule. The new, mechanistic model may provide a rationale for better understanding of digoxin pharmacodynamics and could become a tool to bridge the gap between in vitro and in vivo studies.


Composition influence on pulmonary delivery of rifampicin liposomes.
The effects of lipid concentration and composition on the physicochemical properties, aerosol performance and in vitro toxicity activity of several rifampicin-loaded liposomes were investigated. To this purpose, six liposome formulations containing different amounts of soy phosphatidylcholine and hydrogenated soy phosphatidylcholine, with and without cholesterol and oleic acid, were prepared and fully characterized. Uni- or oligo-lamellar, small (~100 nm), negatively charged (~60 mV) vesicles were obtained. Lipid composition affected aerosol delivery features of liposomal rifampicin; in particular, the highest phospholipid concentration led to a better packing of the vesicular bilayers with a consequent higher nebulization stability. The retention of drug in nebulized vesicles (NER%) was higher for oleic acid containing vesicles (55% ± 1.4%) than for the other samples (~47%). A549 cells were used to evaluate intracellular drug uptake and in vitro toxicity activity of rifampicin-loaded liposomes in comparison with the free drug. Cell toxicity was more evident when oleic acid containing liposomes were used.


In-Pixel Temperature Sensors with an Accuracy of ±0.25 °C, a 3σ Variation of ±0.7 °C in the Spatial Domain and a 3σ Variation of ±1 °C in the Temporal Domain.
This article presents in-pixel (of a CMOS image sensor (CIS)) temperature sensors with improved accuracy in the spatial and the temporal domain. The goal of the temperature sensors is to be used to compensate for dark (current) fixed pattern noise (FPN) during the exposure of the CIS. The temperature sensors are based on substrate parasitic bipolar junction transistor (BJT) and on the nMOS source follower of the pixel. The accuracy of these temperature sensors has been improved in the analog domain by using dynamic element matching (DEM), a temperature independent bias current based on a bandgap reference (BGR) with a temperature independent resistor, correlated double sampling (CDS), and a full BGR bias of the gain amplifier. The accuracy of the bipolar based temperature sensor has been improved to a level of ±0.25 °C, a 3σ variation of ±0.7 °C in the spatial domain, and a 3σ variation of ±1 °C in the temporal domain. In the case of the nMOS based temperature sensor, an accuracy of ±0.45 °C, 3σ variation of ±0.95 °C in the spatial domain, and ±1.4 °C in the temporal domain have been acquired. The temperature range is between -40 °C and 100 °C.


Connections of the mouse subfornical region of the lateral hypothalamus (LHsf).
The lateral hypothalamus is a major integrative hub with a complex architecture characterized by intricate and overlapping cellular populations expressing a large variety of neuro-mediators. In rats, the subfornical lateral hypothalamus (LHsf) was identified as a discrete area with very specific outputs, receiving a strong input from the nucleus incertus, and involved in defensive and foraging behaviors. We identified in the mouse lateral hypothalamus a discrete subfornical region where a conspicuous cluster of neurons express the mu opioid receptor. We thus examined the inputs and outputs of this LHsf region in mice using retrograde tracing with the cholera toxin B subunit and anterograde tracing with biotin dextran amine, respectively. We identified a connectivity profile largely similar, although not identical, to what has been described in rats. Indeed, the mouse LHsf has strong reciprocal connections with the lateral septum, the ventromedial hypothalamic nucleus and the dorsal pre-mammillary nucleus, in addition to a dense output to the lateral habenula. However, the light input from the nucleus incertus and the moderate bidirectional connectivity with nucleus accumbens are specific to the mouse LHsf. A preliminary neurochemical study showed that LHsf neurons expressing mu opioid receptors also co-express calcitonin gene-related peptide or somatostatin and that the reciprocal connection between the LHsf and the lateral septum may be functionally modulated by enkephalins acting on mu opioid receptors. These results suggest that the mouse LHsf may be hodologically and functionally comparable to its rat counterpart, but more atypical connections also suggest a role in consummatory behaviors.


alpha-Mannosidosis: functional cloning of the lysosomal alpha-mannosidase cDNA and identification of a mutation in two affected siblings.
a-Mannosidosis (MIM 248500) is an autosomal recessive lysosomal storage disorder resulting from deficient activity of lysosomal alpha-mannosidase (LAMAN) (EC 220.127.116.11). The disease is characterized by massive intracellular accumulation of mannose-rich oligosaccharides with resulting mental retardation, hearing loss, immune deficiency and skeletal changes. We report here the purification and characterization of human placenta LAMAN. The enzyme is synthesized as a single-chain precursor which is processed into three glycopeptides of 70, 42 and 15 kDa. The 70 kDa peptide is further partially proteolysed into three more peptides that are joined by disulfide bridges. The laman cDNA sequence was assembled from overlapping fragments obtained by PCR on human fibroblast and human lung cDNA. The deduced amino acid sequence contains a putative signal peptide of 48 amino acids followed by a polypeptide sequence of 962 amino acids. Northern blot analyses revealed a single transcript of approximately 3.5 kb present in all tissues examined but at varying levels. Two affected siblings of Palestinian origin were homozygous for a mutation that causes a His-->Leu replacement at a position which is conserved among class 2 alpha-mannosidases from several species.


Can amnesic patients learn without awareness? New evidence comparing deterministic and probabilistic sequence learning.
Can associative learning take place without awareness? We explore this issue in a sequence learning paradigm with amnesic and control participants, who were simply asked to react to one of four possible stimuli on each trial. Unknown to them, successive stimuli occurred in a sequence. We manipulated the extent to which stimuli followed the sequence in a deterministic manner (noiseless condition) or only probabilistically so (noisy condition). Through this paradigm, we aimed at addressing two central issues: first, we asked whether sequence learning takes place in either condition with amnesic patients. Second, we asked whether this learning takes place without awareness. To answer this second question, participants were asked to perform a subsequent sequence generation task under inclusion and exclusion conditions, as well as a recognition task. Reaction times results show that amnesic patients learned the sequence only in the deterministic condition. However, they failed to be able to reproduce the sequence in the generation task. In contrast, we found learning for both sequence structures in control participants, but only control participants exposed to a deterministic sequence were successful in performing the generation task, thus suggesting that the acquired knowledge can be used consciously in this condition. Neither amnesic nor control participants showed correct old/new judgments in the recognition task. The results strengthen the claim that implicit learning is at least partly spared in amnesia, and the role of contextual information available for learning is discussed.


[Description of the psychiatric unit in prisons in the autonomous community of Valencia].
Little work has been done on the contents of psychiatric consultancies in prisons. To explore the sociodemographic, clinical and treatment profile of the patients who were assessed in the psychiatric consultancy of three prisons. This is a prospective, longitudinal, descriptive and multicenter-based epidemiological study of patients who were assessed in the psychiatric consultacy of three prisons in the Autonomous Community of Valencia from 2009 to 2011. Sociodemographic, clinical and prison data was obtained from each patient. Their frequency was compared with Pearson's Ji² and averages with the ANOVA test or with The Kruskall-Wallis method. The probability of keeping the patients in schedule was calculated with Kaplan-Meier's curves and differences with the Mantel-Haenzsel method (Logrank) were established. A logistic regression model was designed to determine the data linked to frequent users. 786 patients were assessed in 2,006 visits (2.5 visits/patient). 90% were male, 88.2% Spanish and their average age was 36 years. 29.9% suffered from an infectious chronic pathology. 69.5% used some kinds of substance. 59.2% suffered from personality disorder and 11.6% from the spectrum of schizophrenia. The most frequent medical profiles were: 49.1% with anxiety disorder, 20.8% depressive disorder and 11.7% psychotic disorder. These psychotic disorders meant a greater probability of 1.5 times for maintaining in schedule. The most commonly prescribed medications were quetiapine, mirtazapine, pregabalin and diazepam. There is a high prevalence of mental disorders in prisons. It is necessary to improve the available resources to deal with these pathologies in the most effective way.


Inferring the spread of COVID-19: the role of time-varying reporting rate in epidemiological modelling.
The role of epidemiological models is crucial for informing public health officials during a public health emergency, such as the COVID-19 pandemic. However, traditional epidemiological models fail to capture the time-varying effects of mitigation strategies and do not account for under-reporting of active cases, thus introducing bias in the estimation of model parameters. To infer more accurate parameter estimates and to reduce the uncertainty of these estimates, we extend the SIR and SEIR epidemiological models with two time-varying parameters that capture the transmission rate and the rate at which active cases are reported to health officials. Using two real data sets of COVID-19 cases, we perform Bayesian inference via our SIR and SEIR models with time-varying transmission and reporting rates and via their standard counterparts with constant rates; our approach provides parameter estimates with more realistic interpretation, and 1-week ahead predictions with reduced uncertainty. Furthermore, we find consistent under-reporting in the number of active cases in the data that we consider, suggesting that the initial phase of the pandemic was more widespread than previously reported.


Compensatory evolution for a gene deletion is not limited to its immediate functional network.
Genetic disruption of an important phenotype should favor compensatory mutations that restore the phenotype. If the genetic basis of the phenotype is modular, with a network of interacting genes whose functions are specific to that phenotype, compensatory mutations are expected among the genes of the affected network. This perspective was tested in the bacteriophage T3 using a genome deleted of its DNA ligase gene, disrupting DNA metabolism. In two replicate, long-term adaptations, phage compensatory evolution accommodated the low ligase level provided by the host without reinventing its own ligase. In both lines, fitness increased substantially but remained well below that of the intact genome. Each line accumulated over a dozen compensating mutations during long-term adaptation, and as expected, many of the compensatory changes were within the DNA metabolism network. However, several compensatory changes were outside the network and defy any role in DNA metabolism or biochemical connection to the disruption. In one line, these extra-network changes were essential to the recovery. The genes experiencing compensatory changes were moderately conserved between T3 and its relative T7 (25% diverged), but the involvement of extra-network changes was greater in T3. Compensatory evolution was only partly limited to the known functionally interacting partners of the deleted gene. Thus gene interactions contributing to fitness were more extensive than suggested by the functional properties currently ascribed to the genes. Compensatory evolution offers an easy method of discovering genome interactions among specific elements that does not rest on an a priori knowledge of those elements or their interactions.


Glucagon-like peptide-1 receptor agonist activation ameliorates venous thrombosis-induced arteriovenous fistula failure in chronic kidney disease.
High shear stress that develops in the arteriovenous fistula of chronic kidney diseases (CKD) may increase H2O2 and thromboxane A2 (TXA2) release, thereby exacerbating endothelial dysfunction, thrombosis, and neointimal hyperplasia. We investigated whether glucagon-like peptide-1 receptor agonist/exendin-4, a potentially cardiovascular protective agent, could improve TXA2-induced arteriovenous fistula injury in CKD. TXA2 administration to H2O2-exposed human umbilical vein endothelial cells increased apoptosis, senescence, and detachment; these phenotypes were associated with the downregulation of phosphorylated endothelial nitric oxide synthase/heme oxygenase-1 (eNOS/HO-1) signalling. Exendin-4 reduced H2O2/TXA2-induced endothelial injury via inhibition of apoptosis-related mechanisms and restoration of phosphorylated eNOS/HO-1 signalling. Male Wistar rats subjected to right common carotid artery-external jugular vein anastomosis were treated with exendin-4 via cervical implant osmotic pumps for 16-42 days. High shear stress induced by the arteriovenous fistula significantly increased venous haemodynamics, blood and tissue H2O2 and TXB2 levels, macrophage/monocyte infiltration, fibrosis, proliferation, and adhesion molecule-1 expression. Apoptosis was also increased due to NADPH oxidase gp91 activation and mitochondrial Bax translocation in the proximal end of the jugular vein of CKD rats. Exendin-4-treatment of rats with CKD led to the restoration of normal endothelial morphology and correction of arteriovenous fistula function. Exendin-4 treatment or thromboxane synthase gene deletion in CKD mice markedly reduced ADP-stimulated platelet adhesion to venous endothelium, and prevented venous occlusion in FeCl3-injured vessels by upregulation of HO-1. Together, these data reveal that the use of glucagon-like peptide-1 receptor agonists is an effective strategy for treatment of CKD-induced arteriovenous fistula failure.


Plant invaders and their novel natural enemies: who is naïve?
Introduced exotic species encounter a wide range of non-coevolved enemies and competitors in their new range. Evolutionary novelty is a key aspect of these interactions, but who benefits from novelty: the exotic species or their new antagonists? Paradoxically, the novelty argument has been used to explain both the release from and the suppression by natural enemies. We argue that this paradox can be solved by considering underlying interaction mechanisms. Using plant defenses as a model, we argue that mismatches between plant and enemy interaction traits can enhance plant invasiveness in the case of toxin-based defenses, whereas invasiveness is counteracted by mismatches in recognition-based defenses and selective foraging of generalist herbivores on plants with rare toxins. We propose that a mechanistic understanding of ecological mismatches can help to explain and predict when evolutionary novelty will enhance or suppress exotic plant invasiveness. This knowledge may also enhance our understanding of plant abundance following range expansion, or during species replacements along successional stages.


Population size and major valleys explain microsatellite variation better than taxonomic units for caribou in western Canada.
Identifying conservation units below the species level is becoming increasingly important, particularly when limited resources necessitate prioritization for conservation among such units. This problem is exemplified with caribou, a mammal with a circum-Arctic distribution that is exposed to a broad spectrum of ecological conditions, but is also declining in many parts of its range. We used microsatellite markers to evaluate the suitability of existing intra-specific taxonomic designations to act as population units for conservation and contrasted this with landscape features that were independent of taxonomy. We also quantified the relationship between genetic differentiation and subpopulation size, a factor that has been under-represented in landscape genetic research. Our data set included three subspecies and three ecotypes of caribou that varied in population size by five orders of magnitude. Our results indicated that genetic structure did not correspond to existing taxonomic designation, particularly at the level of ecotype. Instead, we found that major valleys and population size were the strongest factors associated with substructure. There was a negative exponential relationship between population size and F(ST) between pairs of adjacent subpopulations, suggesting that genetic drift was the mechanism causing the structure among the smallest subpopulations. A genetic assignment test revealed that movement among subpopulations was a fraction of the level needed to stabilize smaller subpopulations, indicating little chance for demographic rescue. Such results may be broadly applicable to landscape genetic studies, because population size and corresponding rates of drift have the potential to confound interpretations of landscape effects on population structure.


Clinical outcome of HIV viraemic controllers and noncontrollers with normal CD4 counts is exclusively determined by antigen-specific CD8+ T-cell-mediated HIV suppression.
In this cross-sectional study we evaluated T-cell responses using several assays to determine immune correlates of HIV control that distinguish untreated viraemic controllers (VC) from noncontrollers (NC) with similar CD4 counts. Samples were taken from 65 ART-naïve chronically HIV-infected VC and NC from Thailand with matching CD4 counts in the normal range (>450 cells/μl). We determined HIVp24-specific T-cell responses using standard Interferon-gamma (IFNγ) ELISpot assays, and compared the functional quality of HIVp24-specific CD8+ T-cell responses using polychromatic flow cytometry. Finally, in vitro HIV suppression assays were performed to evaluate directly the activity of CD8+ T cells in HIV control. Autologous CD4+ T cells were infected with primary patient-derived HIV isolates and the HIV suppressive activity of CD8+ T cells was determined after co-culture, measuring production of HIVp24 Ag by ELISA. The HIVp24-specific T-cell responses of VC and NC could not completely be differentiated through measurement of IFNγ-producing cells using ELISpot assays, nor by the absolute cell numbers of polyfunctional HIVp24-specific CD8+ T cells. However, in vitro HIV suppression assays showed clear differences between VC and NC. HIV suppressive activity, mediated by either ex vivo unstimulated CD8+ T cells or HIVp24-specific T-cell lines, was significantly greater using cells from VC than NC cells. Additionally, we were able to demonstrate a significant correlation between the level of HIV suppressive activity mediated by ex vivo unstimulated CD8+ T cells and plasma viral load (pVL) (Spearman r = -0.7345, p = 0.0003). This study provides evidence that in vitro HIV suppression assays are the most informative in the functional evaluation of CD8+ T-cell responses and can distinguish between VC and NC.


Effects of the COVID-19 pandemic on food insecurity in El Salvador during 2020.
This study sought to quantify the prevalence of food insecurity among Salvadorian households, to identify the determinants of food insecurity and to explore the impact of the COVID-19 pandemic on food insecurity. A nationwide, representative random sample of 2358 households was used for this cross-sectional study. The Household Hunger Scale (HHS) was used to assess the prevalence of food insecurity during a 30-day period. For comparison, three items were used from the Household Food Insecurity Experience Scale (HFIES), which measures hunger occurring during a 12-month time frame. For determinant analysis, binary logistic regression was used for the HHS and ordered logistic regression for the HFIES. The prevalence of food insecurity was 6.45% (152/2356) among Salvadorian households when the HHS was used, affecting 5.48% (129/2356) to a moderate degree and 0.98% (23/2356) to a severe degree. The prevalence significantly increased when the HFIES scale items were used, with 35.41% (835/2358) of households being affected, a figure closer to the national poverty level. Determinants of food insecurity according to the HHS included agricultural problems (P = 0.00, odds ratio [OR] =1.69), the household's prepandemic income (P = 0.00, OR = 0.48) and higher educational levels (i.e. having a secondary education [P = 0.00, OR = 0.31], technical [P = 0.03, OR = 0.24] or university education [P = 0.00, OR = 0.05]). When using the HFIES, the determinants were similar (i.e. income, agricultural problems, educational level). In more than 94% (744/785) of households, participants reported that food insecurity was exacerbated by the COVID-19 pandemic. When compared with other relevant international studies, the prevalence of food insecurity identified using the HHS - only 6.45% - was low for El Salvador. However, when using the HFIES scale, the prevalence rose to 35.41% of households. Some determinants align with previous studies, namely income, educational level and agricultural problems. The COVID-19 pandemic appeared to have direct effects on food insecurity.


Long-lasting subjective effects of LSD in normal subjects.
Lysergic acid diethylamide (LSD) and other serotonergic hallucinogens can induce profound alterations of consciousness and mystical-type experiences, with reportedly long-lasting effects on subjective well-being and personality. We investigated the lasting effects of a single dose of LSD (200 μg) that was administered in a laboratory setting in 16 healthy participants. The following outcome measures were assessed before and 1 and 12 months after LSD administration: Persisting Effects Questionnaire (PEQ), Mysticism Scale (MS), Death Transcendence Scale (DTS), NEO-Five Factor Inventory (NEO-FFI), and State-Trait Anxiety Inventory (STAI). On the PEQ, positive attitudes about life and/or self, positive mood changes, altruistic/positive social effects, positive behavioral changes, and well-being/life satisfaction significantly increased at 1 and 12 months and were subjectively attributed by the subjects to the LSD experience. Five-Dimensions of Altered States of Consciousness (5D-ASC) total scores, reflecting acutely induced alterations in consciousness, and Mystical Experience Questionnaire (MEQ30) total scores correlated with changes in well-being/life satisfaction 12 months after LSD administration. No changes in negative attitudes, negative mood, antisocial/negative social effects, or negative behavior were attributed to the LSD experience. After 12 months, 10 of 14 participants rated their LSD experience as among the top 10 most meaningful experiences in their lives. Five participants rated the LSD experience among the five most spiritually meaningful experiences in their lives. On the MS and DTS, ratings of mystical experiences significantly increased 1 and 12 months after LSD administration compared with the pre-LSD screening. No relevant changes in personality measures were found. In healthy research subjects, the administration of a single dose of LSD (200 μg) in a safe setting was subjectively considered a personally meaningful experience that had long-lasting subjective positive effects. Registration identification number: NCT01878942.


cDNA-AFLP analysis of gene expression differences between the flower bud and sprout-shoot apical meristem of Angelica sinensis (Oliv.) Diels.
Angelica sinensis (Oliv.) Diels (Umbelliferae) is a well-known medicinal plant mainly distributed in Gansu Province of China. Its local and global demand is significant because of its food and medicinal applications. However, the early bolting rate of Angelica sinensis (Oliv.) Diels reaches 20%-60%, which seriously affects its food and medicinal qualities. Thus, differences in gene expression between the flower bud and sprout-shoot apical meristem underwent analysis, by means of cDNA-amplified restriction fragment length polymorphism, to better understand the flowering mechanism. 64 primer sets, each of which amplified to 60 transcript-derived fragments (TDFs), were used. Among these TDFs, 26 were expressed specifically in the flower bud. After cloning and sequencing, 32 distinct sequences were obtained from these 26 TDFs, and 25 were found with homologous sequences in databases. Confirmation of differential expression of 13 sequences was obtained by semi-quantitative RT-PCR, their showing higher expression levels in flower buds. These homologous sequences encode transposable elements, pentatricopeptide repeat-containing proteins, DNA-binding transcription factors, zinc finger (B-box type) family proteins, NADP-dependent sorbitol 6-phosphate dehydrogenase (S6PDH), amongst others.


Lack of change in neurochemical markers during the postepileptic phase of intrahippocampal tetanus toxin syndrome in rats.
The chronic epileptic syndrome induced by injecting tetanus toxin into rat hippocampus causes functional changes that essentially are permanent, outlasting the period of active seizures by at least 1 year. These long-term changes have been characterized by an impaired performance on a range of behavioral tasks, which in turn have been associated with a physiologic depression of hippocampal evoked responses but not with any discernible histopathology. In the present study, we examined the hippocampi of rats in the postseizure phase of the tetanus toxin model and observed no significant changes in the concentration of neurochemical markers for six neurotransmitters. Therefore, the long-term reduction in hippocampal excitability cannot be attributed to any major loss of afferents or hippocampal neurons using aspartate, acetylcholine, gamma-aminobutyric acid (GABA), glutamate, norepinephrine (NE), or serotonin as their transmitters.


Validity of Real-Time Data Generated by a Wearable Microtechnology Device.
The purpose of this study was to investigate the validity of global positioning system (GPS) and micro-electrical-mechanical-system (MEMS) data generated in real time through a dedicated receiver. Postsession data acted as the criterion as it is used to plan the volume and intensity of future training and is downloaded directly from the device. Twenty-five professional rugby league players completed 2 training sessions wearing an MEMS device (Catapult S5, firmware version: 5.27). During sessions, real-time data were collected through the manufacturer receiver and dedicated software (Openfield v1.14), which was positioned outdoors at the same location for every session. The GPS variables included total-, low- (0-3 m·s), moderate- (3.1-5 m·s), high- (5.1-7 m·s), and very high-speed (>7.1 m·s) distances. Micro-electrical-mechanical-system data included total session PlayerLoad. When compared to postsession data, mean bias for total-, low-, moderate-, high-, and very high-speed distances were all trivial, with the typical error of the estimate (TEE) small, small, trivial, trivial and small, respectively. Pearson correlation coefficients for total-, low-, moderate-, high- and very-high-speed distances were nearly perfect, nearly perfect, perfect, perfect, and nearly perfect, respectively. For PlayerLoad, mean bias was trivial, whereas TEE was moderate and correlation nearly perfect. Practitioners should be confident that when interpreting real-time speed-derived metrics, the data generated in real-time are comparable with those downloaded directly from the device postsession. However, practitioners should refrain from interpreting accelerometer-derived data (i.e., PlayerLoad) or acknowledge the moderate error associated with this real-time measure.


Life style related risk factors for cardiovascular disease among patients at a teaching hospital in Karachi.
Cardiovascular disease is a leading cause for morbidity and mortality all over the world. It is important to study life style related, modifiable cardiovascular risk factors among patients, in order to devise preventive strategies. We surveyed family practice patients visiting the out-patient clinics of Aga Khan University Hospital, Karachi. The interview was questionnaire-based and recorded the demographic profile of the patients, in addition to information on life style related cardiovascular risk factors. The ethical requirements for the study were met. SPSS computer software was used for data management. Fifty patients were surveyed and included 28 (56%) men and 22 (44%) women. Thirty seven (74%) respondents were married, nineteen (38%) had graduate education, twenty five (50%) were in private service, and twelve (24%) were housewives. Twenty six (52%) respondents were overweight or obese. Butter, cream, margarine and red meat was consumed by 17 (34%) and 32 (64%) respondents respectively. White meat, fresh fruits and vegetables were consumed by 30 (60%), 29 (58%) and 38 (76%) respondents respectively. Oil from vegetable sources was used by 49 (98%) respondents. Addition of table salt was found occasionally and always in 18 (36%) and 09 (18%) respondents respectively. Twenty seven (54%) respondents exercised at least twenty minutes, three times a week, while eleven (22%) were current smokers. Life style related modifiable risk factors are prevalent in patients reporting to our hospital. This offers an opportunity for preventive strategies, to prevent cardiovascular disease. Such surveys, followed by interventional strategies, are recommended to be followed by all Primary care facilities.


Gastric juice for the diagnosis of H pylori infection in patients on proton pump inhibitors.
To determine the efficacy of gastric juice polymerase chain reaction (PCR) for the detection of H pylori infection in comparison with histology and gastric antral biopsy PCR in patients on a proton pump inhibitor (PPI). Eighty-five consecutive patients with dyspeptic symptoms were enrolled. Gastric biopsies for histology, PCR and gastric juice were collected at endoscopy for PCR of the H pylori urease C gene (ure C). Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy, positive and negative likelihood ratio for PCR of gastric juice for the H pylori ure C gene was compared to histology and gastric antral biopsy H pylori ure C PCR in patients with and without PPI. Gastric juice PCR was positive in 66 (78%) patients. Histology showed H pylori associated gastritis in 57 (67%). Gastric biopsy PCR was positive in 72 (85%). In patients not taking PPI, the sensitivity, specificity, PPV, NPV, accuracy and positive and negative likelihood ratio for gastric juice PCR were 89%, 72%, 91%, 67%, 90%, 85%, 3.1 and 0.1 respectively. In patients on PPI these values were 86%, 100%%, 100%, 29%, 86%, 9.5 and 1.4, respectively. Gastric juice PCR for the diagnosis of H pylori infection has increased sensitivity compared to histology with PPI. The use of gastric juice PCR is recommended to confirm H pylori status in patients taking PPIs.


First-Principles Investigation of Nanopore Sequencing Using Variable Voltage Bias on Graphene-Based Nanoribbons.
In this study, we examine the mechanism of nanopore-based DNA sequencing using a voltage bias across a graphene nanoribbon. Using density function theory and a nonequilibrium Green's function approach, we determine the transmission spectra and current profile for adenine, guanine, cytosine, thymine, and uracil as a function of bias voltage in an energy minimized configuration. Utilizing the transmission current, we provide a general methodology for the development of a three nanopore graphene-based device that can be used to distinguish between the various nucleobases for DNA/RNA sequencing. From our analysis, we deduce that it is possible to use different transverse currents across a multinanopore device to differentiate between nucleobases using various voltages of 0.5, 1.3, and 1.6 V. Overall, our goal is to improve nanopore design to further DNA/RNA nucleobase sequencing and biomolecule identification techniques.


Predictors of mortality in hemodialysis patients.
Serum biochemical measures suggestive of undernutrition have been reported to correlate with 1-yr mortality risk in prevalent groups of hemodialysis patients. The predictive power of these variables has not been reported in newly diagnosed patients or in patients whose dialysis prescription is guided by urea kinetics. The relationship of these predictors to mortality over periods of longer than 1 yr is also unreported. Therefore, the survival of 184 hemodialysis patients was examined for up to 44 months (1987 to 1991) with the Cox proportional hazards model. Baseline demographic, clinical, and biochemical parameters were used as independent variables. To adjust for bias in patient selection, the survival of patients with 12 months or less of prior dialysis at the time of enrollment ("new cases") was analyzed separately from that of patients with more than 1 yr of prior treatment ("long-standing cases"). Serum albumin was less than 3.5 g/dL in 31% of new cases and in 12% of long-standing cases. Adjusting for the other variables, low serum albumin was the strongest mortality risk predictor in both new and long-standing cases. Low serum cholesterol was an independent risk predictor in both groups. Diabetes and race were not significant predictors. Mean age at enrollment was nearly a decade higher for nonsurvivors than for survivors, in both new and long-standing groups. Yet, age was not an independent risk predictor in the Cox model for the new group because of an unexpectedly high death rate among young black men. Female gender, which was confounded by increased age, took the place of age in the model for the new group. For each model, there was good agreement between observed and predicted mortality for up to 24 months. To assess the influence of dialysis treatment time and dose (measured as pre-to-post treatment urea ratio) on risk, survival was examined in a subset of 139 patients monitored for up to 22 months, from 1989 to 1991, a period when the urea ratio was used routinely. Adjusting for the other variables, low serum albumin and cholesterol again independently increased risk. The urea ratio was also a significant independent predictor. The pattern of mortality by urea ratio was U shaped, with minimum risk for values between 2.5 and 3.4 Treatment time did not influence risk. It was concluded that baseline serum values of albumin and cholesterol strongly influence survival for up to 2 yr in new and long-standing hemodialysis patients.(ABSTRACT TRUNCATED AT 400 WORDS)


The ghosts of departed quantities: approaches to dealing with observations below the limit of quantitation.
A common but not necessarily logical requirement in drug development is that a 'limit of quantitation' be set for chemical assays and that observations that fall below the limit should not be treated as real data but should be labelled as below the limit and set aside for special treatment. We examine five of seven approaches to analysing such data considered by Beal in 2001, concentrating in particular on two: one that treats the data as a truncated sample and another that treats them as a censored sample. In fact, using a pattern-mixture framework, one can show that the former consists of using the conditional distribution of the 'acceptable values' and the latter adds the information from the marginal mixing distribution. We illustrate these approaches with a real example, concentrating in particular on the two likelihood-based methods, provide various formulae that may be used to compare these and other approaches, check these formulae using simulations and make some recommendations as to which approach one should use.


Indoor allergens, environmental avoidance, and allergic respiratory disease.
Indoor allergen exposure to sources such as house-dust mites, pets, fungi, and insects plays a significant role in patients with allergic rhinitis and asthma. The identification of the major allergens has led to methods that can quantitate exposure, e.g., immunoassays for Der p 1 in settled dust samples. Sensitization and the development of allergic respiratory disease result from complex genetic and environmental interactions. New paradigms that examine the role of other environmental factors, including exposure to proteases that can activate eosinophils and initiate Th2 responses, and epigenetics, are being explored. Recommendations for specific environmental allergen avoidance measures are discussed for house-dust mites, cockroaches, animal dander, and fungi. Specific measures to reduce indoor allergen exposure when vigorously applied may reduce the risk of sensitization and symptoms of allergic respiratory disease, although further research will be necessary to establish cost-effective approaches.


Impact of prenatal diagnosis by ultrasound on the prevalence of congenital anomalies at birth in southern France.
The aims were (1) to assess whether termination of pregnancy after prenatal screening by ultrasound affected the prevalence of congenital anomalies at birth, and (2) to examine the trend of this pattern over time. This study deals with congenital anomalies, possibly detectable prenatally or at birth, which were classified as isolated and multiple anomalies; chromosomal anomalies were not included. The prevalence rates of congenital anomalies at birth were determined from case registration data in the Marseille district, France, from the registry of congenital malformations (Eurocat no 22), which covers 23,500 births a year. The chi 2 test for homogeneity in proportions was used to test whether the differences in the total prevalence rates were significant over time. The population was defined as all children born to parents living in the Marseille district between January 1 1984 and December 31 1990. Among the 164,509 pregnancy outcomes monitored during the study, 1795 children with a single congenital anomaly and 288 with multiple congenital anomalies detectable at birth were assessed. The percentage of pregnancy terminations was higher in the case of multiple anomalies (16%) than with single ones (7.5%). Leaving aside the lethal birth defects, this percentage became 7.9% in the case of multiple anomalies and 4.3% with isolated ones. A significant increase (p < 0.001) occurred over the seven year study period in the total percentage of terminations because of isolated anomalies but not in that involving multiple ones. The increase observed in the former case was found to be mainly attributable to an increase in the number of terminations of pregnancy undertaken because of anomalies which were either lethal or associated with very low survival rates (p < 0.001). Termination of pregnancy after prenatal ultrasound examination was found to have a definite impact on the prevalence at birth of lethal and congenital anomalies with a low survival rate, and this impact tended to increase over time. No such impact was observed in the case of congenital anomalies associated with high survival rates.


Polysubstance use in treatment seekers who inject amphetamine: Drug use profiles, injecting practices and quality of life.
The injection of amphetamine is becoming increasingly common. However, there has been a lack of research examining people who inject amphetamine as the primary drug of use, limiting the potential to ensure services address the unique needs of this group. The current study used latent class analysis to identify classes of polydrug use among people who report injecting amphetamine during the past 12months. It also examined differences between classes and drug use patterns, injecting practices, quality of life and psychological distress. Participants who were attending non-government specialist alcohol and other drug treatment across New South Wales, Australia and had identified amphetamine as their principle drug of concern and reported injecting amphetamine in the previous 12months were included in the current study (N=827). Latent class analysis was performed to identify polydrug profiles of participants. The large majority of people in the current study (85%) demonstrated low probability of heroin or other opiate use. Three distinct classes of polydrug use were identified: (1) Low-polydrug (n=491), (2) Opiates-polydrug (n=123), and (3) Alcohol-polydrug (n=213). There was a trend for the Low-polydrug class to demonstrate better functioning and safer injecting practices than the Opiates-polydrug and Alcohol-polydrug classes. The results suggest that the majority of people accessing treatment who inject amphetamine as their primary drug of choice have a low probability of heroin or other opiate use. It is important that future research consider whether traditional harm minimisation strategies are appropriate for people who primarily inject amphetamine.


Testosterone treatment is not associated with increased risk of prostate cancer or worsening of lower urinary tract symptoms: prostate health outcomes in the Registry of Hypogonadism in Men.
To evaluate the effects of testosterone-replacement therapy (TRT) on prostate health indicators in hypogonadal men, including rates of prostate cancer diagnoses, changes in prostate-specific antigen (PSA) levels and lower urinary tract symptoms (LUTS) over time. The Registry of Hypogonadism in Men (RHYME) is a multi-national patient registry of treated and untreated, newly-diagnosed hypogonadal men (n = 999). Follow-up assessments were performed at 3-6, 12, 24, and 36 months. Baseline and follow-up data collection included medical history, physical examination, blood sampling, and patient questionnaires. Prostate biopsies underwent blinded independent adjudication for the presence and severity of prostate cancer; PSA and testosterone levels were measured via local and central laboratory assays; and LUTS severity was assessed via the International Prostate Symptom Score (IPSS). Incidence rates per 100 000 person-years were calculated. Longitudinal mixed models were used to assess effects of testosterone on PSA levels and IPSS. Of the 999 men with clinically diagnosed hypogonadism (HG), 750 (75%) initiated TRT, contributing 23 900 person-months of exposure. The mean testosterone levels increased from 8.3 to 15.4 nmol/L in treated men, compared to only a slight increase from 9.4 to 11.3 nmol/L in untreated men. In all, 55 biopsies were performed for suspected prostate cancer, and 12 non-cancer related biopsies were performed for other reasons. Overall, the proportion of positive biopsies was nearly identical in men on TRT (37.5%) compared to those not on TRT (37.0%) over the course of the study. There were no differences in PSA levels, total IPSS, or the IPSS obstructive sub-scale score by TRT status. Lower IPSS irritative sub-scale scores were reported in treated compared to untreated men. Results support prostate safety of TRT in newly diagnosed men with HG.


Evaluation of rs1957106 Polymorphism of NF-κBI in Glioblastoma Multiforme in Isfahan, Iran.
The kB family of nuclear factor (NF-κB) is a series of transcription factors that plays a key role in regulation of immunity, cell growth, and apoptosis and is considered as the main downstream component of epidermal growth factor receptor for which there are evidence of excessive activity in most cases of glioblastoma multiform (GBM). Thus, the current information has gained evidence on NF-κBIA tumor suppressor role in GMB. SNP rs1957106 was diagnosed as a new polymorphism which affected the expression of NF-κBI and causes activation of NF-κB in GBM patients. This study was conducted on 100 cases of GBM including 47 paraffin-embedded brain tissue samples and 53 blood samples from another 53 GBM patients and 150 controls. The NF-κBI rs1957106 SNP was identified by the NCBI, and genotyping was performed by high-resolution melt (HRM) assay. Melt curves from HRM which suspected to single-nucleotide polymorphism (SNP) were selected and subjected to direct sequencing. The distribution of allele A of NF-κβ gene in patients with GBM with 31% was not significantly different from healthy participants (27.3%) (P = 0.375). Furthermore, the distribution of AG and GG genotypes in comparison with AA genotypes did not show a significant correlation with GBM incidence (P > 0.05). Findings of the present study provide evidence that the rs1957106 SNP in NF-κBIA is found more in GBM patients, but it was not statistically significant. As there are conflicting studies showing significant higher rate of this SNP in GBM, further study is suggested.


Sepiapterin Enhances Tumor Radio- and Chemosensitivities by Promoting Vascular Normalization.
Previously, we demonstrated that nitric oxide (NO) synthase (NOS) is uncoupled in a wide range of solid tumors and that restoring NOS coupling with the tetrahydrobiopterin precursor sepiapterin (SP) inhibits tumor progression. Endothelial dysfunction characterizes the poorly functional vasculature of solid tumors, and since NO is critical for regulation of endothelial function we asked whether SP, by recoupling NOS, improves tumor vasculature structure and function-enhancing chemotherapeutic delivery and response to radiotherapy. MMTV-neu mice with spontaneous breast tumors were treated with SP by oral gavage and evaluated by multispectral optoacoustic tomographic analysis of tumor HbO2 and by tissue staining for markers of hypoxia, blood perfusion, and markers of endothelial and smooth muscle proteins. Recoupling tumor NOS activity results in vascular normalization observed as reduced tumor hypoxia, improved tumor percentage of HbO2 and perfusion, as well as increased pericyte coverage of tumor blood vessels. The normalized vasculature and improved tumor oxygenation led to a greater than 2-fold increase in radiation-induced apoptosis compared with radiation or SP alone. High-performance liquid chromatography analysis of tumor doxorubicin levels showed a greater than 50% increase in doxorubicin uptake and a synergistic effect on tumor cell apoptosis. This study highlights for the first time the importance of NOS uncoupling and endothelial dysfunction in the development of tumor vasculature and presents a new approach for improving the tumoricidal efficacies of chemotherapy and radiotherapy.


Outer membrane protein A (OmpA) contributes to serum resistance and pathogenicity of Escherichia coli K-1.
We examined whether outer membrane protein A (OmpA) contributes to gram-negative pathogenesis by determining the effect of mutagenesis of ompA in a virulent Escherichia coli K-1 isolate. An OmpA mutant was generated by insertion of the transposon TnphoA, which was genetically modified to increase the efficiency of its delivery by conjugation. The mutant was less virulent than its parent strain in two models of E. coli K-1 infection. Equal inocula of the OmpA+ and OmpA- strains fed to neonatal rats resulted in a sevenfold-greater incidence of bacteremia at 72 h from the OmpA+ strain. The lethal effect of the OmpA- mutant was significantly less than that of the OmpA+ parent strain when inoculated onto the chorioallantoic membrane of 10-day embryonated chick eggs. There was, however, no difference between strains in growth characteristics under physiologic conditions, either in rat serum or in unembryonated chick eggs. In the presence of a 10-day chick embryo, there was a 10-fold increase in the survival and growth of the OmpA+ strain. Correction of the mutation in ompA with an E. coli K-12 ompA gene restored a level of virulence equivalent to that of the parent strain. The ompA mutant was more sensitive to the bactericidal effect of pooled human serum by the classical pathway of complement activation. These results suggest that OmpA contributes to E. coli K-1 pathogenesis by a mechanism which may involve increased serum resistance.


Effect of roasted pea flour/starch and encapsulated pea starch incorporation on the in vitro starch digestibility of pea breads.
Oven or microwave roasting and alginate encapsulation of pea flour and starch to produce novel pea ingredients for enrichment of slowly digestible starch (SDS) and resistant starch (RS) content in pea bread were investigated. Pea flour treated either by oven roasting (160°C, 30min) or by microwave roasting (1.1kW, 6min) effectively retained its low starch digestibility similar to its native form (∼25% SDS; ∼60% RS). When oven roasting was applied to pea starch, SDS content increased triply compared to the fully boiled counterpart. Alginate encapsulation effectively controlled carbohydrate release to simulated gastric, intestinal and colonic fluids, and thus largely enriched the SDS and RS fractions in starch. Pea bread containing up to 37.5% of encapsulated roasted MPS pea starch not only provided high SDS and RS fractions (23.9% SDS and 30.2% RS) compared to a white bread control (0.2% SDS and 2.5% RS), but also provided an acceptable palatability.


IgE cross-reactivity between meadow fescue pollen and kiwi fruit in patients' sera with sensitivity to both extracts.
The presence of IgE reactivity to kiwi fruit and grass pollen allergens which could be caused by cross-reactivity has been detected in many patients with allergy. Proper identification of allergens as well as cross-reactive components is essential for understanding fruit- and pollen-associated hypersensitivity. Using the sera from the polysensitized patients with specific IgE to grass pollen and kiwi fruit we tested reactivity to both allergen sources. IgE reactivity was exhibited in 8 serum samples by immunoblot. A serum pool formed by 8 individual sera was used for the investigation of IgE crossreactivity. SDS-PAGE immunoblot-inhibition assay was performed by preincubation of the sera with meadow fescue pollen, kiwi fruit extract, and isolated 24 kDa kiwi protein. To determine the allergens of kiwi fruit extract, we performed 2D PAGE immunoblot. In order to detect the crossreactive components between two allergen sources, a specific IgE for the 24 kDa kiwi allergen was purified. SDS-PAGE immunoblot meadow fescue pollen showed allergens ranging from 94 to 16 kDa, and kiwi fruit had 12 allergens ranging from 94 to 17 kDa. 2D-PAGE analysis revealed at least 15 spots in the kiwi extract and about 10 allergens. The most prominent allergen in 2D PAGE immunoblot was protein with 24 kDa and pI 9.4-9.5. Using an affinity-purified specific IgE we found that the 24 kDa kiwi allergen shared IgE-reactive epitopes with the meadow fescue group 4 and allergen about 36 kDa. Crossreactivity between isolated 24 kDa kiwi allergen and Fes p 4 was confirmed by anti-grass group 4 moab 2D8. Our findings showed that fescue meadow pollen cross-sensitize to kiwi fruits. A 24 kDa kiwi glycoprotein represent potential major allergen, which share common epitopes with Fes p 4 and 36 kDa meadow fescue allergen.


Short-term effects of nasal proportional assist ventilation in patients with chronic hypercapnic respiratory insufficiency.
Proportional assist ventilation (PAV) has recently been proposed as a mode of synchronized partial ventilatory support. This study evaluates the short-term effects of nasal PAV on arterial blood gases in stable patients with chronic hypercapnia. Forty two patients (30 with chronic obstructive pulmonary disease (COPD) and 12 with restrictive chest wall disease (RCWD) due to kyphoscoliosis) underwent a 1 h run of nasal PAV. Randomly, two levels of assistance were performed: 1) PAV was set at a level corresponding to volume assist (VA) and flow assist (FA) at 80% of the individual values of elastance (Ers) and resistance (Rrs) obtained with the "runaway" method; and 2) VA and FA were set at a value corresponding to the difference between the patients' individual Ers and Rrs and normal values of Ers and Rrs. Arterial blood gases and dyspnoea (by visual analogue scale (VAS)) were evaluated in all patients during unsupported ventilation and 60 min of PAV. PAV was well tolerated and resulted in significant improvement in arterial oxygen tension (Pa,O2), arterial carbon dioxide tension (Pa,CO2) (6.8+/-0.8 to 7.4+/-1.4 and 7.2/-0.9 to 6.8+/-0.9 kPa, respectively) and VAS (29+/-23 to 20+/-18%). The effects of PAV were not different in the two groups of diseases nor in the two groups of settings. Different settings of nasal proportional assist ventilation are well tolerated and may improve gas exchange and dyspnoea in patients with stable hypercapnic respiratory insufficiency.


MicroRNA-126-mediated control of cell fate in B-cell myeloid progenitors as a potential alternative to transcriptional factors.
Lineage specification is thought to be largely regulated at the level of transcription, where lineage-specific transcription factors drive specific cell fates. MicroRNAs (miR), vital to many cell functions, act posttranscriptionally to decrease the expression of target mRNAs. MLL-AF4 acute lymphocytic leukemia exhibits both myeloid and B-cell surface markers, suggesting that the transformed cells are B-cell myeloid progenitor cells. Through gain- and loss-of-function experiments, we demonstrated that microRNA 126 (miR-126) drives B-cell myeloid biphenotypic leukemia differentiation toward B cells without changing expression of E2A immunoglobulin enhancer-binding factor E12/E47 (E2A), early B-cell factor 1 (EBF1), or paired box protein 5, which are critical transcription factors in B-lymphopoiesis. Similar induction of B-cell differentiation by miR-126 was observed in normal hematopoietic cells in vitro and in vivo in uncommitted murine c-Kit(+)Sca1(+)Lineage(-) cells, with insulin regulatory subunit-1 acting as a target of miR-126. Importantly, in EBF1-deficient hematopoietic progenitor cells, which fail to differentiate into B cells, miR-126 significantly up-regulated B220, and induced the expression of B-cell genes, including recombination activating genes-1/2 and CD79a/b. These data suggest that miR-126 can at least partly rescue B-cell development independently of EBF1. These experiments show that miR-126 regulates myeloid vs. B-cell fate through an alternative machinery, establishing the critical role of miRNAs in the lineage specification of multipotent mammalian cells.


Cardiopulmonary resuscitation: the science behind the hands.
Sudden cardiac arrest is a leading cause of death worldwide. Despite significant advances in resuscitation science since the initial use of external chest compressions in humans nearly 60 years ago, there continues to be wide variability in rates of successful resuscitation across communities. The American Heart Association (AHA) and European Resuscitation Council emphasise the importance of high-quality chest compressions as the foundation of resuscitation care. We review the physiological basis for the association between chest compression quality and clinical outcomes and the scientific basis for the AHA's key metrics for high-quality cardiopulmonary resuscitation. Finally, we highlight that implementation of strategies that promote effective chest compressions can improve outcomes in all patients with cardiac arrest.


The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase.
The silent information regulator 2 protein (Sir2p) of Saccharomyces cerevisiae is an NAD-dependent histone deacetylase that plays a critical role in transcriptional silencing. Here, we report that a human ortholog of Sir2p, sirtuin type 2 (SIRT2), is a predominantly cytoplasmic protein that colocalizes with microtubules. SIRT2 deacetylates lysine-40 of alpha-tubulin both in vitro and in vivo. Knockdown of SIRT2 via siRNA results in tubulin hyperacetylation. SIRT2 colocalizes and interacts in vivo with HDAC6, another tubulin deacetylase. Enzymatic analysis of recombinant SIRT2 in comparison to a yeast homolog of Sir2 protein (Hst2p) shows a striking preference of SIRT2 for acetylated tubulin peptide as a substrate relative to acetylated histone H3 peptide. These observations establish SIRT2 as a bona fide tubulin deacetylase.


Artificial Fusion of mCherry Enhances Trehalose Transferase Solubility and Stability.
LeLoir glycosyltransferases are important biocatalysts for the production of glycosidic bonds in natural products, chiral building blocks, and pharmaceuticals. Trehalose transferase (TreT) is of particular interest since it catalyzes the stereo- and enantioselective α,α-(1→1) coupling of a nucleotide sugar donor and monosaccharide acceptor for the synthesis of disaccharide derivatives. Heterologously expressed thermophilic trehalose transferases were found to be intrinsically aggregation prone and are mainly expressed as catalytically active inclusion bodies in Escherichia coli To disfavor protein aggregation, the thermostable protein mCherry was explored as a fluorescent protein tag. The fusion of mCherry to trehalose transferase from Pyrobaculum yellowstonensis (PyTreT) demonstrated increased protein solubility. Chaotropic agents like guanidine or the divalent cations Mn(II), Ca(II), and Mg(II) enhanced the enzyme activity of the fusion protein. The thermodynamic equilibrium constant, Keq, for the reversible synthesis of trehalose from glucose and a nucleotide sugar was determined in both the synthesis and hydrolysis directions utilizing UDP-glucose and ADP-glucose, respectively. UDP-glucose was shown to achieve higher conversions than ADP-glucose, highlighting the importance of the choice of nucleotide sugars for LeLoir glycosyltransferases under thermodynamic control.IMPORTANCE The heterologous expression of proteins in Escherichia coli is of great relevance for their functional and structural characterization and applications. However, the formation of insoluble inclusion bodies is observed in approximately 70% of all cases, and the subsequent effects can range from reduced soluble protein yields to a complete failure of the expression system. Here, we present an efficient methodology for the production and analysis of a thermostable, aggregation-prone trehalose transferase (TreT) from Pyrobaculum yellowstonensis via its fusion with mCherry as a thermostable fluorescent protein tag. This fusion strategy allowed for increased enzyme stability and solubility and could be applied to other (thermostable) proteins, allowing rapid visualization and quantification of the mCherry-fused protein of interest. Finally, we have demonstrated that the enzymatic synthesis of trehalose from glucose and a nucleotide sugar is reversible by approaching the thermodynamic equilibrium in both the synthesis and hydrolysis directions. Our results show that uridine establishes an equilibrium constant which is more in favor of the product trehalose than when adenosine is employed as the nucleotide under identical conditions. The influence of different nucleotides on the reaction can be generalized for all LeLoir glycosyltransferases under thermodynamic control as the position of the equilibrium depends solely on the reaction conditions and is not affected by the nature of the catalyst.


Zoledronic acid impairs myeloid differentiation to tumour-associated macrophages in mesothelioma.
Suppressive immune cells present in tumour microenvironments are known to augment tumour growth and hamper efficacy of antitumour therapies. The amino-bisphosphonate Zoledronic acid (ZA) is considered as an antitumour agent, as recent studies showed that ZA prolongs disease-free survival in cancer patients. The exact mechanism is a topic of debate; it has been suggested that ZA targets tumour-associated macrophages (TAMs). We investigate the role of ZA on the myeloid differentiation to TAMs in murine mesothelioma in vivo and in vitro. Mice were intraperitoneally inoculated with a lethal dose of mesothelioma tumour cells and treated with ZA to determine the effects on myeloid differentiation and survival. We show that ZA impaired myeloid differentiation. Inhibition of myeloid differentiation led to a reduction in TAMs, but the number of immature myeloid cells with myeloid-derived suppressor cell (MDSC) characteristics was increased. In addition, ZA affects the phenotype of macrophages leading to reduced level of TAM-associated cytokines in the tumour microenvironment. No improvement of survival was observed. We conclude that ZA leads to a reduction in macrophages and impairs polarisation towards an M2 phenotype, but this was associated with an increase in the number of immature myeloid cells, which might diminish the effects of ZA on survival.


Hospital admission for selected single virus infections prior to diabetes mellitus.
To determine whether hospital admission for a range of specified virus infections was followed by a raised admission rate for diabetes mellitus; and, if raised, whether the increase is compatible with the hypothesis that virus infection is a cause of diabetes. Analysis of a database of hospital statistics including admissions for people with diabetes mellitus before the age of 30 years. There was no evidence of excess risk of diabetes after measles, mumps, rubella, infectious mononucleosis, influenza, infectious hepatitis, varicella and herpes zoster, herpes simplex, aseptic meningitis or bronchiolitis. For example, of 1433 patients admitted for measles, 6 were later admitted with diabetes (risk ratio 1.32; 95% confidence interval 0.5-2.9); of 866 patients admitted for mumps, 2 were later admitted for diabetes (risk ratio 0.74; 0.1-2.7). Numbers of people with diabetes subsequent to infection were too few, however, to rule out the possibility of small effects. Our findings do not support the hypothesis that any of these virus infections initiate the processes that lead to the development of diabetes, or that these infections act as a trigger to precipitate active disease in those whose diabetes is already present but latent.


Comparisons of Differentiation Potential in Human Mesenchymal Stem Cells from Wharton's Jelly, Bone Marrow, and Pancreatic Tissues.
Background. Type 1 diabetes mellitus results from autoimmune destruction of β-cells. Insulin-producing cells (IPCs) differentiated from mesenchymal stem cells (MSCs) in human tissues decrease blood glucose levels and improve survival in diabetic rats. We compared the differential ability and the curative effect of IPCs from three types of human tissue to determine the ideal source of cell therapy for diabetes. Methods. We induced MSCs from Wharton's jelly (WJ), bone marrow (BM), and surgically resected pancreatic tissue to differentiate into IPCs. The in vitro differential function of these IPCs was compared by insulin-to-DNA ratios and C-peptide levels after glucose challenge. In vivo curative effects of IPCs transplanted into diabetic rats were monitored by weekly blood glucose measurement. Results. WJ-MSCs showed better proliferation and differentiation potential than pancreatic MSCs and BM-MSCs. In vivo, WJ-IPCs significantly reduced blood glucose levels at first week after transplantation and maintained significant decrease till week 8. BM-IPCs reduced blood glucose levels at first week but gradually increased since week 3. In resected pancreas-IPCs group, blood glucose levels were significantly reduced till two weeks after transplantation and gradually increased since week 4. Conclusion. WJ-MSCs are the most promising stem cell source for β-cell regeneration in diabetes treatment.


Respiratory toxicity of buprenorphine results from the blockage of P-glycoprotein-mediated efflux of norbuprenorphine at the blood-brain barrier in mice.
Deaths due to asphyxia as well as following acute poisoning with severe respiratory depression have been attributed to buprenorphine in opioid abusers. However, in human and animal studies, buprenorphine exhibited ceiling respiratory effects, whereas its metabolite, norbuprenorphine, was assessed as being a potent respiratory depressor in rodents. Recently, norbuprenorphine, in contrast to buprenorphine, was shown in vitro to be a substrate of human P-glycoprotein, a drug-transporter involved in all steps of pharmacokinetics including transport at the blood-brain barrier. Our objectives were to assess P-glycoprotein involvement in norbuprenorphine transport in vivo and study its role in the modulation of buprenorphine-related respiratory effects in mice. University-affiliated research laboratory, INSERM U705, Paris, France. Wild-type and P-glycoprotein knockout female Friend virus B-type mice. Respiratory effects were studied using plethysmography and the P-glycoprotein role at the blood-brain barrier using in situ brain perfusion. Norbuprenorphine(≥ 1 mg/kg) and to a lesser extent buprenorphine (≥ 10 mg/kg) were responsible for dose-dependent respiratory depression combining increased inspiratory (TI) and expiratory times (TE). PSC833, a powerful P-glycoprotein inhibitor, significantly enhanced buprenorphine-related effects on TI (p < .01) and TE (p < .05) and norbuprenorphine-related effects on minute volume (VE, p < .05), TI, and TE (p < .001). In P-glycoprotein-knockout mice, buprenorphine-related effects on VE (p < .01), TE (p < .001), and TI (p < .05) and norbuprenorphine-related effects on VE (p < .05) and TI (p < .001) were significantly enhanced. Plasma norbuprenorphine concentrations were significantly increased in PSC833-treated mice (p < .001), supporting a P-glycoprotein role in norbuprenorphine pharmacokinetics. Brain norbuprenorphine efflux was significantly reduced in PSC833-treated and P-glycoprotein-knockout mice (p < .001), supporting P-glycoprotein-mediated norbuprenorphine transport at the blood-brain barrier. P-glycoprotein plays a key-protective role in buprenorphine-related respiratory effects, by allowing norbuprenorphine efflux at the blood-brain barrier. Our findings suggest a major role for drug-drug interactions that lead to P-glycoprotein inhibition in buprenorphine-associated fatalities and respiratory depression.


Just another case of diarrhea and vomiting?
Chronic intussusception is a treatable cause of failure to thrive. It is an uncommon condition often diagnosed late. The presenting features differ from acute intussusception. Chronic intussusception presents with a varying combination of abdominal pain, vomiting, weight loss/failure to thrive, diarrhea, and blood per rectum. An abdominal mass may or may not be palpable. The classic triad of abdominal pain, vomiting, and blood per rectum is uncommon. The purpose of this report is to present a small series of 3 cases and review 19 previously reported cases of chronic intussusception. Considering the diagnosis of chronic intussusception in children who present with failure to thrive and recurrent nonspecific abdominal symptoms is emphasized.


Helicobacter pylori infection and prevalence of stroke.
Helicobacter pylori causes peptic ulcer disease; however, conflicting evidence exists regarding its role in extragastric conditions. We aimed to examine associations of H pylori infection and peptic ulcer disease with stroke. A cross-sectional study was undertaken using data of 147 936 individuals aged 25-95 years who underwent the urea breath test during 2002-2012, based on the computerized database of the second largest health maintenance organization in Israel. Logistic regression models were fitted to control for potential confounders. Overall, 1397 (0.9%) patients had stroke and 76 965 (52.0%) had a H pylori positive test. The likelihood of prevalent stroke increased in relation to H pylori infection: adjusted odds ratio (aOR) 1.16 (95% confidence intervals [CI]: 1.04-1.29), gastric ulcer: aOR 1.50 (95% CI: 1.18-1.91), and duodenal ulcer: aOR 1.25 (95% CI: 1.07-1.46). The results support the premise that stroke may be associated with a history of H pylori infection.


Innate immune activation in intestinal homeostasis.
Loss of intestinal immune regulation leading to aberrant immune responses to the commensal microbiota are believed to precipitate the chronic inflammation observed in the gastrointestinal tract of patients with inflammatory bowel diseases (IBD), Crohn's disease and ulcerative colitis. Innate immune receptors that recognize conserved components derived from the microbiota are widely expressed by both epithelial cells and leucocytes of the gastrointestinal tract and play a key role in host protection from infectious pathogens; yet precisely how pathogenic and commensal microbes are distinguished is not understood. Furthermore, aberrant innate immune activation may also drive intestinal pathology, as patients with IBD exhibit extensive infiltration of innate immune cells to the inflamed intestine, and polymorphisms in many innate immunity genes influence susceptibility to IBD. Thus, a balanced interaction between the microbiota and innate immune activation is required to maintain a healthy mutualistic relationship between the microbiota and the host, which when disturbed can result in intestinal inflammation.


Generalist care managers for the treatment of depressed medicaid patients in North Carolina: a pilot study.
In most states, mental illness costs are an increasing share of Medicaid expenditures. Specialized depression care managers (CM) have consistently demonstrated improvements in patient outcomes relative to usual primary care (UC), but are costly and may not be fully utilized in smaller practices. A generalist care manager (GCM) could manage multiple chronic conditions and be more accepted and cost-effective than the specialist depression CM. We designed a pilot program to demonstrate the feasibility of training/deploying GCMs into primary care settings. We randomized depressed adult Medicaid patients in 2 primary care practices in Western North Carolina to a GCM intervention or to UC. GCMs, already providing services in diabetes and asthma in both study arms, were further trained to provide depression services including self-management, decision support, use of information systems, and care management. The following data were analyzed: baseline, 3- and 6-month Patient Health Questionnaire (PHQ9) scores; baseline and 6-month Short Form (SF) 12 scores; Medicaid claims data; questionnaire on patients' perceptions of treatment; GCM case notes; physician and office staff time study; and physician and office staff focus group discussions. Forty-five patients were enrolled, the majority with preexisting depression. Both groups improved; the GCM group did not demonstrate better clinical and functional outcomes than the UC group. Patients in the GCM group were more likely to have prescriptions of correct dosing by chart data. GCMs most often addressed comorbid conditions (36%), then social issues (27%) and appointment reminders (14%). GCMs recorded an average of 46 interactions per patient in the GCM arm. Focus group data demonstrated that physicians valued using GCMs. A time study documented that staff required no more time interacting with GCMs, whereas physicians spent an average of 4 minutes more per week. GCMs can be trained in care of depression and other chronic illnesses, are acceptable to practices and patients, and result in physicians prescribing guideline concordant care. GCMs appear to be a feasible intervention for community medical practices and to warrant a larger scale trial to test their appropriateness for Medicaid programs nationally.


Immunity in amyotrophic lateral sclerosis: blurred lines between excessive inflammation and inefficient immune responses.
Despite wide genetic, environmental and clinical heterogeneity in amyotrophic lateral sclerosis, a rapidly fatal neurodegenerative disease targeting motoneurons, neuroinflammation is a common finding. It is marked by local glial activation, T cell infiltration and systemic immune system activation. The immune system has a prominent role in the pathogenesis of various chronic diseases, hence some of them, including some types of cancer, are successfully targeted by immunotherapeutic approaches. However, various anti-inflammatory or immunosuppressive therapies in amyotrophic lateral sclerosis have failed. This prompted increased scrutiny over the immune-mediated processes underlying amyotrophic lateral sclerosis. Perhaps the biggest conundrum is that amyotrophic lateral sclerosis pathogenesis exhibits features of three otherwise distinct immune dysfunctions-excessive inflammation, autoimmunity and inefficient immune responses. Epidemiological and genome-wide association studies show only minimal overlap between amyotrophic lateral sclerosis and autoimmune diseases, so excessive inflammation is usually thought to be secondary to protein aggregation, mitochondrial damage or other stresses. In contrast, several recently characterized amyotrophic lateral sclerosis-linked mutations, including those in TBK1, OPTN, CYLD and C9orf72, could lead to inefficient immune responses and/or damage pile-up, suggesting that an innate immunodeficiency may also be a trigger and/or modifier of this disease. In such cases, non-selective immunosuppression would further restrict neuroprotective immune responses. Here we discuss multiple layers of immune-mediated neuroprotection and neurotoxicity in amyotrophic lateral sclerosis. Particular focus is placed on individual patient mutations that directly or indirectly affect the immune system, and the mechanisms by which these mutations influence disease progression. The topic of immunity in amyotrophic lateral sclerosis is timely and relevant, because it is one of the few common and potentially malleable denominators in this heterogenous disease. Importantly, amyotrophic lateral sclerosis progression has recently been intricately linked to patient T cell and monocyte profiles, as well as polymorphisms in cytokine and chemokine receptors. For this reason, precise patient stratification based on immunophenotyping will be crucial for efficient therapies.


[Memory and aging: changes in the mammillary body and anterior thalamic nuclei due to age].
Karyometric changes and variation of neuron number in mammillary body and anterior thalamic nuclei, dependent of the age, have been studied in humans and rats. The behaviour of the neurons number is very similar in all studied nuclei of both species: there is a constant and gradual loss of neurons from the first until the last period of the life. The intensity of the neuronal loss, however, is different according the nuclei and is more pronounced in rats than in humans. The evolution of the nuclear area of the human mammillary body is similar to that of the anterior thalamic nuclei: there is a significant decrease until the age of 60-70 years and then begins an increase up to the end of the life. This increase is more intense in the mammillary nuclei than in the thalamic ones. In rats, the behaviour of the mammillary nuclei is different to that of the anterior thalamic nuclei. In the mammillary body there is an increase of the nuclear are until 18 months of age, followed by a decrease until the end of the life. The anterior thalamic nuclei show a double inflexion: first, it appears a marked decrease of the nuclear area during the first 6 months, then the area remains unchanged until the age of 15 months, but from this period up to the end of the 2nd year undergoes an increase and, finally, during the 3 year shows a significant decrease. There is not a complete correlation between neuronal loss and nuclear area increase. Although the neuronal loss is constant during the entire life, the nuclear are increase only in a certain period: in rats during the second year and in humans in the final period of life. The signification of this partial correlation is discussed.


Breeding objectives for sheep should be customised depending on variation in pasture growth across years.
Breeding programmes for livestock require economic weights for traits that reflect the most profitable animal in a given production system, which affect the response in each trait after selection. The profitability of sheep production systems is affected by changes in pasture growth as well as grain, meat and wool prices between seasons and across years. Annual pasture growth varies between regions within Australia's Mediterranean climate zone from low growth with long periods of drought to high growth with shorter periods of drought. Therefore, the objective of this study was to assess whether breeding objectives need to be adapted for regions, depending on how reliable the pasture growth is across years. We modelled farms with Merino sheep bred for wool and meat in 10 regions in Western Australia. Across these 10 regions, mean annual pasture growth decreased, and the CV of annual pasture growth increased as pasture growth for regions became less reliable. We calculated economic values for nine traits, optimising management across 11 years, including variation for pasture growth and wool, meat and grain prices between and within years from 2002 to 2012. These economic values were used to calculate responses to selection for each trait for the 10 regions. We identified two potential breeding objectives, one for regions with low or high reliability and the other for regions with medium reliability of pasture growth. Breeding objectives for high or low pasture growth reliability had more emphasis on live weight traits and number of lambs weaned. Breeding objectives for medium reliability of pasture growth had more emphasis on decreasing fibre diameter. Relative economic weights for fleece weight did not change across the regions. Regions with low or high pasture reliability had similar breeding objectives and response to selection, because the relationship between the economic values and CV of pasture growth were not linear for live weight traits and the number of lambs weaned. This non-linearity was caused by differences in distribution of pasture growth between regions, particularly during summer and autumn, when ewes were pregnant, with increases in energy requirements affecting the value of lambs weaned. In addition, increasing live weight increased the intake capacity of sheep, which meant that more poor quality pasture could be consumed during summer and autumn, which had more value in regions with low and high pasture reliability. We concluded that breeding values for sheep production systems should be customised depending on the reliability of pasture growth between years.


Expression and nucleotide sequence of the Lactobacillus bulgaricus beta-galactosidase gene cloned in Escherichia coli.
The Lactobacillus bulgaricus beta-galactosidase gene was cloned on a ca. 7-kilobase-pair HindIII fragment in the vector pKK223-3 and expressed in Escherichia coli by using its own promoter. The nucleotide sequence of the gene and approximately 400 bases of 3'- and 5'-flanking sequences was determined. The amino acid sequence of the beta-galactosidase, deduced from the nucleotide sequence of the gene, yielded a monomeric molecular mass of ca. 114 kilodaltons, slightly smaller than the E. coli lacZ and Klebsiella pneumoniae lacZ enzymes but larger than the E. coli evolved (ebgA) beta-galactosidase. The cloned beta-galactosidase was found to be indistinguishable from the native enzyme by several criteria. From amino acid sequence alignments, the L. bulgaricus beta-galactosidase has a 30 to 34% similarity to the E. coli lacZ, E. coli ebgA, and K. pneumoniae lacZ enzymes. There are seven regions of high similarity common to all four of these beta-galactosidases. Also, the putative active-site residues (Glu-461 and Tyr-503 in the E. coli lacZ beta-galactosidase) are conserved in the L. bulgaricus enzyme as well as in the other two beta-galactosidases mentioned above. The conservation of active-site amino acids and the large regions of similarity suggest that all four of these beta-galactosidases evolved from a common ancestral gene. However, these enzymes are quite different from the thermophilic beta-galactosidase encoded by the Bacillus stearothermophilus bgaB gene.


Age mediation of frontoparietal activation during visual feature search.
Activation of frontal and parietal brain regions is associated with attentional control during visual search. We used fMRI to characterize age-related differences in frontoparietal activation in a highly efficient feature search task, detection of a shape singleton. On half of the trials, a salient distractor (a color singleton) was present in the display. The hypothesis was that frontoparietal activation mediated the relation between age and attentional capture by the salient distractor. Participants were healthy, community-dwelling individuals, 21 younger adults (19-29 years of age) and 21 older adults (60-87 years of age). Top-down attention, in the form of target predictability, was associated with an improvement in search performance that was comparable for younger and older adults. The increase in search reaction time (RT) associated with the salient distractor (attentional capture), standardized to correct for generalized age-related slowing, was greater for older adults than for younger adults. On trials with a color singleton distractor, search RT increased as a function of increasing activation in frontal regions, for both age groups combined, suggesting increased task difficulty. Mediational analyses disconfirmed the hypothesized model, in which frontal activation mediated the age-related increase in attentional capture, but supported an alternative model in which age was a mediator of the relation between frontal activation and capture.


Cancer and palliative care in COVID-19 and other challenging situations-highlights from the Uganda Cancer Institute-Palliative Care Association of Uganda 3rd Uganda Conference on Cancer and Palliative Care, 23-24 September 2021, held in Kampala, Uganda and virtually.
The 3rd Uganda Conference on Cancer and Palliative Care was held in September 2021 with the theme: cancer and palliative care in COVID-19 and other challenging situations. It was hosted by the Uganda Cancer Institute and the Palliative Care Association of Uganda (UCI-PCAU). The conference was held virtually, with a mix of pre-recorded sessions, plenary sessions being broadcast live on television (TV) by the Uganda Broadcasting Corporation TV, live speakers at the studio and others presenting in real time via Zoom. The conference brought together >350 participants who participated on Zoom, along with those attending in person at the studio and those watching the plenary sessions on TV. At the heart of this joint UCI-PCAU conference was the commitment to not only continue but to improve the provision of cancer care and palliative care within Uganda. Key themes from the conference included: the importance of Universal Health Coverage; the impact of COVID-19 on the provision of cancer and palliative care; that both cancer care and palliative care are available in Uganda; education for all; the importance of working together to provide care and overcome challenges, e.g. through technology; the resilience shown by those working in cancer and palliative care; the grief experienced by so many people who have lost loved ones during the pandemic; the importance of good health seeking behaviour - prevention is better than cure; the challenge of funding; the need for health care equity for marginalised and vulnerable populations and finally we can't wait for the world to stop COVID-19 - COVID-19 is here to stay - we need to find solutions. The last few years have seen significant challenges due to the COVID-19 pandemic; however, despite this, cancer and palliative care service provision has continued. This conference, whilst unique and very different from previous conferences, was a great opportunity to share not only amongst each other, but also to share key messages with the public through the live broadcasting of the plenary sessions of the conference.


Variability of neural activation during walking in humans: short heels and big calves.
People come in different shapes and sizes. In particular, calf muscle size in humans varies considerably. One possible cause for the different shapes of calf muscles is the inherent difference in neural signals sent to these muscles during walking. In sedentary adults, the variability in neural control of the calf muscles was examined with muscle size, walking kinematics and limb morphometrics. Half the subjects walked while activating their medial gastrocnemius (MG) muscles more strongly than their lateral gastrocnemius (LG) muscles during most walking speeds ('MG-biased'). The other subjects walked while activating their MG and LG muscles nearly equally ('unbiased'). Those who walked with an MG-biased recruitment pattern also had thicker MG muscles and shorter heel lengths, or MG muscle moment arms, than unbiased walkers, but were similar in height, weight, lower limb length, foot length, and exhibited similar walking kinematics. The relatively less plastic skeletal system may drive calf muscle size and motor recruitment patterns of walking in humans.


Location Isn't Everything: Proximity, Hospital Characteristics, Choice of Hospital, and Disparities for Breast Cancer Surgery Patients.
Assess the relative importance of proximity and other hospital characteristics in the choice of hospital for breast cancer surgery by race/ethnicity. SEER-Medicare data. Observational study of women aged >65 years receiving surgery for stage I/II/III breast cancer diagnosed in 1992-2007 in Detroit (N = 10,746 white/black), Atlanta (N = 4,018 white/black), Los Angeles (N = 9,433 white/black/Asian/Hispanic), and San Francisco (N = 4,856 white/black/Asian). We calculated the distance from each patient's census tract of residence to each area hospital. We estimated discrete choice models for the probability of receiving surgery at each hospital based on distance and assessed whether deviations from these predictions entailed interactions of hospital characteristics with the patient's race/ethnicity. We identified high-quality hospitals by rates of adjuvant radiation therapy and by survey measures of patient experiences, and we assessed how observed surgery rates at high-quality hospitals deviated from those predicted based on distance alone. Proximity was significantly associated with hospital choice in all areas. Minority more often than white breast cancer patients had surgery at hospitals with more minority patients, those treating more Medicaid patients, and in some areas, lower quality hospitals. Residential location alone does not explain concentration of racial/ethnic-minority breast cancer surgery patients in certain hospitals that are sometimes of lower quality.


A kick-start for CLC antiporters' pharmacology.
Despite the role of chloride channels and anion/proton antiporters of the CLC protein family in physiological processes and different genetic diseases, their pharmacology has been under-developed. In this issue of Chemistry & Biology, Howery et al. report the synthesis of 4'-octanamidostilbene-2,2'-disulfonate, the first high-affinity inhibitor of a CLC antiporter, a critical step toward reviving the CLC pharmacology.


Empirically evaluating the WHO global code of practice on the international recruitment of health personnel's impact on four high-income countries four years after adoption.
Shortages of health workers in low-income countries are exacerbated by the international migration of health workers to more affluent countries. This problem is compounded by the active recruitment of health workers by destination countries, particularly Australia, Canada, UK and USA. The World Health Organization (WHO) adopted a voluntary Code of Practice in May 2010 to mitigate tensions between health workers' right to migrate and the shortage of health workers in source countries. The first empirical impact evaluation of this Code was conducted 11-months after its adoption and demonstrated a lack of impact on health workforce recruitment policy and practice in the short-term. This second empirical impact evaluation was conducted 4-years post-adoption using the same methodology to determine whether there have been any changes in the perceived utility, applicability, and implementation of the Code in the medium-term. Forty-four respondents representing government, civil society and the private sector from Australia, Canada, UK and USA completed an email-based survey evaluating their awareness of the Code, perceived impact, changes to policy or recruitment practices resulting from the Code, and the effectiveness of non-binding Codes generally. The same survey instrument from the original study was used to facilitate direct comparability of responses. Key lessons were identified through thematic analysis. The main findings between the initial impact evaluation and the current one are unchanged. Both sets of key informants reported no significant policy or regulatory changes to health worker recruitment in their countries as a direct result of the Code due to its lack of incentives, institutional mechanisms and interest mobilizers. Participants emphasized the existence of previous bilateral and regional Codes, the WHO Code's non-binding nature, and the primacy of competing domestic healthcare priorities in explaining this perceived lack of impact. The Code has probably still not produced the tangible improvements in health worker flows it aspired to achieve. Several actions, including a focus on developing bilateral codes, linking the Code to topical global priorities, and reframing the Code's purpose to emphasize health system sustainability, are proposed to improve the Code's uptake and impact.


Regeneration of the radial nerve cord in the sea cucumber Holothuria glaberrima.
Regeneration of neurons and fibers in the mammalian spinal cord has not been plausible, even though extensive studies have been made to understand the restrictive factors involved. New experimental models and strategies are necessary to determine how new nerve cells are generated and how fibers regrow and connect with their targets in adult animals. Non-vertebrate deuterostomes might provide some answers to these questions. Echinoderms, with their amazing regenerative capacities could serve as model systems; however, very few studies have been done to study the regeneration of their nervous system. We have studied nerve cord regeneration in the echinoderm Holothuria glaberrima. These are sea cucumbers or holothurians members of the class Holothuroidea. One radial nerve cord, part of the echinoderm CNS, was completely transected using a scalpel blade. Animals were allowed to heal for up to four weeks (2, 6, 12, 20, and 28 days post-injury) before sacrificed. Tissues were sectioned in a cryostat and changes in the radial nerve cord were analyzed using classical dyes and immunohistochemistry. In addition, the temporal and spatial distribution of cell proliferation and apoptosis was assayed using BrdU incorporation and the TUNEL assay, respectively.We found that H. glaberrima can regenerate its radial nerve cord within a month following transection. The regenerated cord looks amazingly similar in overall morphology and cellular composition to the uninjured cord. The cellular events associated to radial cord regeneration include: (1) outgrowth of nerve fibers from the injured radial cord stumps, (2) intense cellular division in the cord stumps and in the regenerating radial nerve cords, (3) high levels of apoptosis in the RNC adjacent to the injury and within the regenerating cord and (4) an increase in the number of spherule-containing cells. These events are similar to those that occur in other body wall tissues during wound healing and during regeneration of the intestine. Our data indicate that holothurians are capable of rapid and complete regeneration of the main component of their CNS. Regeneration involves both the outgrowth of nerve fibers and the formation of neurons. Moreover, the cellular events employed during regeneration are similar to those involved in other regenerative processes, namely wound healing and intestinal regeneration. Thus, holothurians should be viewed as an alternative model where many of the questions regarding nervous system regeneration in deuterostomes could be answered.