Shared Cultural History as a Predictor of Political and Economic Changes among Nation States.
Political and economic risks arise from social phenomena that spread within and across countries. Regime changes, protest movements, and stock market and default shocks can have ramifications across the globe. Quantitative models have made great strides at predicting these events in recent decades but incorporate few explicitly measured cultural variables. However, in recent years cultural evolutionary theory has emerged as a major paradigm to understand the inheritance and diffusion of human cultural variation. Here, we combine these two strands of research by proposing that measures of socio-linguistic affiliation derived from language phylogenies track variation in cultural norms that influence how political and economic changes diffuse across the globe. First, we show that changes over time in a country's democratic or autocratic character correlate with simultaneous changes among their socio-linguistic affiliations more than with changes of spatially proximate countries. Second, we find that models of changes in sovereign default status favor including socio-linguistic affiliations in addition to spatial data. These findings suggest that better measurement of cultural networks could be profoundly useful to policy makers who wish to diversify commercial, social, and other forms of investment across political and economic risks on an international scale.


Origins of the parasitophorous vacuole membrane of the malaria parasite, Plasmodium falciparum, in human red blood cells.
We have attempted to determine whether the parasitophorous vacuole membrane, in which the malaria parasite (merozoite) encapsulates itself when it enters a red blood cell, is derived from the host cell plasma membrane, as the appearance of the invasion process in the electron microscope has been taken to suggest, or from lipid material stored in the merozoite. We have incorporated into the red cell membrane a haptenic phospholipid, phosphatidylethanolamine, containing an NBD (N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)) group, substituted in the acyl chain, and allowed it to translocate into the inner bilayer leaflet. After invasion of these labelled cells by the parasite, Plasmodium falciparum, immuno-gold electron microscopy was used to follow the distribution of the labelled lipid; this was found to be overwhelmingly in favour of the host cell membrane relative to the parasitophorous vacuole. Merozoites of P. knowlesi were allowed to attach irreversibly to red cells without invasion, using the method of pretreatment with cytochalasin. The region of contact between the merozoite and the host cell membrane was in all cases devoid of the labelled phosphatidylethanolamine. These results lead us to infer that the parasitophorous vacuole membrane is derived wholly or partly from lipid preexisting in the merozoite.


Preoperative Risk Score to Predict Occult Metastatic or Locally Advanced Disease in Patients with Resectable Perihilar Cholangiocarcinoma on Imaging.
Many patients with resectable perihilar cholangiocarcinoma (PHC) on imaging are diagnosed intraoperatively with occult metastatic or locally advanced disease, precluding a curative-intent resection. This study aimed to develop and validate a preoperative risk score. Patients with resectable PHC on imaging who underwent operations in 2 high-volume centers (US and Europe) between 2000 and 2015 were included. Multivariable logistic regression analysis was used to develop the risk score. Cross-validation was used to validate the score, alternating the 2 centers as "training" and "testing" datasets. Of 566 patients who underwent operations, 309 (55%) patients had a resection, and in 257 (45%) patients, a curative-intent resection was precluded due to distant metastasis (n = 151 [27%]) or locally advanced disease (n = 106 [19%]). Preoperative predictors included bilirubin >2 mg/dL, bile duct involvement on imaging, portal vein involvement on imaging (≥180 degrees), hepatic artery involvement on imaging (≥180 degrees), and suspicious lymph nodes on imaging. The new risk score (c-index 0.75 after cross-validation) provided significantly more accurate predictions than the Bismuth classification (c-index 0.62), Blumgart T-staging (c-index 0.67), and cTNM staging (c-index 0.68). The new risk score identified 4 risk groups for occult metastatic or locally advanced disease: low (14.7%), intermediate (29.5%), high (47.3%), and very high risk (81.3%). The preoperative score groups also predicted survival after operation, irrespective of intraoperative findings (p < 0.001). The validated risk score can predict occult distant metastatic or locally advanced PHC based on 5 preoperatively available factors. The score can be useful in preoperative shared decision making and selection of patients in neoadjuvant clinical trials.


Ubiquitin binding by a variant Jab1/MPN domain in the essential pre-mRNA splicing factor Prp8p.
The U1, U2, U4/U6, and U5 small nuclear ribonucleoproteins (snRNPs) are components of the spliceosome, which catalyzes pre-mRNA splicing. One of the largest and the most highly conserved proteins in the spliceosome is Prp8p, a component of the U5 snRNP. Despite its size and conservation, very few motifs have been identified that suggest specific biochemical functions. A variant of the Jab1/MPN domain found in a class of deubiquitinating enzymes is present near the C terminus of Prp8p. Ubiquitination regulates a broad range of cellular pathways, and its functions generally require ubiquitin recognition by one or more ubiquitin-binding domains (UBDs). No precise role for ubiquitin has been defined in the pre-mRNA splicing pathway, and no known UBDs have been found within splicing proteins. Here we show that a Prp8p fragment containing the Jab1/MPN domain binds directly to ubiquitin with an affinity comparable to other known UBDs. Several mutations within this domain that compromise splicing also reduce interaction of the fragment with ubiquitin-Sepharose. Our results define a new UBD and suggest functional links between ubiquitin and the pre-mRNA splicing machinery.


The effect of aging on the pharmacokinetics of nizatidine.
A single oral dose of 300 mg of nizatidine was administered to two groups of volunteers: a first group of 12 young healthy subjects and a second group of 12 elderly patients, in order to compare the pharmacokinetic parameters and to verify a possible influence of the age on nizatidine's kinetics. Blood samples were collected periodically and plasma concentrations of the drug were determined by a specific HPLC method. The results evidenced differences among the pharmacokinetic parameters of the two groups: the mean of Cmax was higher in the elderly than in the young (3.28 mg/ml against 2.23 mg/ml), the peak time was prolonged in the elderly (2.1 h against 1.00 h) and the AUC values were higher too (12.17 in the elderly and 7.99 in the young). Only the elimination half-life t1/2 was slightly higher in the elderly (1.8 h against 1.7 h in the young).


Renewable poly(δ-decalactone) based block copolymer micelles as drug delivery vehicle: in vitro and in vivo evaluation.
Polymers from natural resources are attracting much attention in various fields including drug delivery as green alternatives to fossil fuel based polymers. In this quest, novel block copolymers based on renewable poly(δ-decalactone) (PDL) were evaluated for their drug delivery capabilities and compared with a fossil fuel based polymer i.e. methoxy-poly(ethylene glycol)-b-poly(ε-caprolactone) (mPEG-b-PCL). Using curcumin as a hydrophobic drug model, micelles of PDL block copolymers with different orientation i.e. AB (mPEG-b-PDL), ABA (PDL-b-PEG-b-PDL), ABC (mPEG-b-PDL-b-poly(pentadecalactone) and (mPEG-b-PCL) were prepared by nanoprecipitation method. The size, drug loading and curcumin stability studies results indicated that mPEG-b-PDL micelles was comparable to its counterpart mPEG-b-PCL micelles towards improved delivery of curcumin. Therefore, mixed micelles using these two copolymers were also evaluated to see any change in size, loading and drug release. Drug release studies proposed that sustained release can be obtained using poly(pentadecalactone) as crystalline core whereas rapid release can be achieved using amorphous PDL core. Further, mPEG-b-PDL micelles were found to be non-haemolytic, up to the concentration of 40 mg/mL. In vivo toxicity studies on rats advised low-toxic behaviour of these micelles up to 400 mg/kg dose, as evident by histopathological and biochemical analysis. In summary, it is anticipated that mPEG-b-PDL block copolymer micelles could serve as a renewable alternative for mPEG-b-PCL copolymers in drug delivery applications.


The cardiovascular effect of the uremic solute indole-3 acetic acid.
In CKD, uremic solutes may induce endothelial dysfunction, inflammation, and oxidative stress, leading to increased cardiovascular risk. We investigated whether the uremic solute indole-3 acetic acid (IAA) predicts clinical outcomes in patients with CKD and has prooxidant and proinflammatory effects. We studied 120 patients with CKD. During the median study period of 966 days, 29 patients died and 35 experienced a major cardiovascular event. Kaplan-Meier analysis revealed that mortality and cardiovascular events were significantly higher in the higher IAA group (IAA>3.73 µM) than in the lower IAA group (IAA<3.73 µM). Multivariate Cox regression analysis demonstrated that serum IAA was a significant predictor of mortality and cardiovascular events after adjustments for age and sex; cholesterol, systolic BP, and smoking; C-reactive protein, phosphate, body mass index, and albumin; diastolic BP and history of cardiovascular disease; and uremic toxins p-cresyl sulfate and indoxyl sulfate. Notably, IAA level remained predictive of mortality when adjusted for CKD stage. IAA levels were positively correlated with markers of inflammation and oxidative stress: C-reactive protein and malondialdehyde, respectively. In cultured human endothelial cells, IAA activated an inflammatory nongenomic aryl hydrocarbon receptor (AhR)/p38MAPK/NF-κB pathway that induced the proinflammatory enzyme cyclooxygenase-2. Additionally, IAA increased production of endothelial reactive oxygen species. In conclusion, serum IAA may be an independent predictor of mortality and cardiovascular events in patients with CKD. In vitro, IAA induces endothelial inflammation and oxidative stress and activates an inflammatory AhR/p38MAPK/NF-κB pathway.


Major advances in fundamental dairy cattle nutrition.
Fundamental nutrition seeks to describe the complex biochemical reactions involved in assimilation and processing of nutrients by various tissues and organs, and to quantify nutrient movement (flux) through those processes. Over the last 25 yr, considerable progress has been made in increasing our understanding of metabolism in dairy cattle. Major advances have been made at all levels of biological organization, including the whole animal, organ systems, tissues, cells, and molecules. At the whole-animal level, progress has been made in delineating metabolism during late pregnancy and the transition to lactation, as well as in whole-body use of energy-yielding substrates and amino acids for growth in young calves. An explosion of research using multicatheterization techniques has led to better quantitative descriptions of nutrient use by tissues of the portal-drained viscera (digestive tract, pancreas, and associated adipose tissues) and liver. Isolated tissue preparations have provided important information on the interrelationships among glucose, fatty acid, and amino acid metabolism in liver, adipose tissue, and mammary gland, as well as the regulation of these pathways during different physiological states. Finally, the last 25 yr has witnessed the birth of "molecular biology" approaches to understanding fundamental nutrition. Although measurements of mRNA abundance for proteins of interest already have provided new insights into regulation of metabolism, the next 25 yr will likely see remarkable advances as these techniques continue to be applied to problems of dairy cattle biology. Integration of the "omics" technologies (functional genomics, proteomics, and metabolomics) with measurements of tissue metabolism obtained by other methods is a particularly exciting prospect for the future. The result should be improved animal health and well being, more efficient dairy production, and better models to predict nutritional requirements and provide rations to meet those requirements.


Understanding Time-driven Activity-based Costing.
Transitioning to a value-based health care system will require providers to increasingly scrutinize their outcomes and costs. Although there has been a great deal of effort to understand outcomes, cost accounting in health care has been a greater challenge. Currently the cost accounting methods used by hospitals and providers are based off a fee-for-service system. As resources become increasingly scarce and the health care system attempts to understand which services provide the greatest value, it will be critically important to understand the true costs of delivering a service. An understanding of the true costs of a particular service will help providers make smarter decisions on how to allocate and utilize resources as well as determine which activities are nonvalue added. Achieving value will require providers to have a greater focus on accurate outcome data as well as better methods of cost accounting.


Limited Polymorphism of the Kelch Propeller Domain in Plasmodium malariae and P. ovale Isolates from Thailand.
Artemisinin resistance in Plasmodium falciparum, the agent of severe malaria, is currently a major obstacle to malaria control in Southeast Asia. A gene named "kelch13" has been associated with artemisinin resistance in P. falciparum The orthologue of the kelch gene in P. vivax was identified and a small number of mutations were found in previous studies. The kelch orthologues in the other two human malaria parasites, P. malariae and P. ovale, have not yet been studied. Therefore, in this study, the orthologous kelch genes of P. malariae, P. ovale wallikeri, and P. ovale curtisi were isolated and analyzed for the first time. The homologies of the kelch genes of P. malariae and P. ovale were 84.8% and 82.7%, respectively, compared to the gene in P. falciparum kelch polymorphisms were studied in 13 P. malariae and 5 P. ovale isolates from Thailand. There were 2 nonsynonymous mutations found in these samples. One mutation was P533L, which was found in 1 of 13 P. malariae isolates, and the other was K137R, found in 1 isolate of P. ovale wallikeri (n = 4). This result needs to be considered in the context of widespread artemisinin used within the region; their functional consequences for artemisinin sensitivity in P. malariae and P. ovale will need to be elucidated.


A New Conceptual Approach for Voice Feminization: 12 Years of Experience.
Voice feminization has been developed with various surgical techniques to increase voice pitch. The purpose of this study was to evaluate 12 years of experience using a new endoscopical surgical technique, vocal fold shortening and retrodisplacement of the anterior commissure (VFSRAC) and retraining the voice pattern performed in female patients with androphonia or male-to-female transsexuals. Retrospective study. A retrospective study was performed on 362 patients who underwent VFSRAC (34 with androphonia, eight with androgenital syndrome, four with aplastic anemia treated with androgens, three with pseudohermaphroditism, and 313 male-to-female [MTF] transsexuals) and phonatory pattern rehabilitation between 2003 and 2014. Subjective and perceptual assessments, aerodynamic and acoustic assessments, and a videostroboscopic assessment were evaluated pre- and postoperatively in the female androphonia group and the MTF transsexual group. Mean preoperative fundamental frequency (F0 ) was 144.1 Hz, and mean postoperative F0 was 207.4 Hz, 6 months postoperatively. The mean increase of F0 in the female androphonia group was 53.0 Hz but was 73.6 Hz in the MTF transsexual group after the postoperative phonatory pattern retraining program. Voice femininity increased on the postoperative subjective assessment. Acoustic and aerodynamic assessments were within the normal range. Regularity of the vocal fold mucosal wave were maintained in the normal range. These findings suggest that patients could achieve a natural phonation pattern after surgery. Our new approach for voice feminization, VFSRAC, and retraining of the phonatory pattern was effective for patients who wanted a natural feminine voice. 4 Laryngoscope, 127:1102-1108, 2017.


Fungicide Sensitivity of Sclerotinia homoeocarpa from Golf Courses in Ohio.
Managing dollar spot, the most common and chronic disease on intensively cultivated turfgrass, relies on the judicious use of fungicides. The heavy use of fungicides has led to the development of isolates of Sclerotinia homoeocarpa insensitive to several classes of fungicides, including benzimidazoles, demethylation-inhibitors, and dicarboximides. In vitro fungicide sensitivity assays using single discriminatory concentrations of thiophanate-methyl, propiconazole, and iprodione were developed in this study for evaluating field efficacy of these fungicides and the prevalence of fungicide insensitivity within S. homoeocarpa isolated from golf courses throughout Ohio. Discriminatory concentrations for these fungicides were determined to be: thiophanate-methyl = 1,000 μg a.i. ml-1, propiconazole = 0.1 μg a.i. ml-1, and iprodione = 1.0 μg a.i. ml-1. Effective concentration that produces 50% inhibition (EC50) was estimated based on relative mycelial growth of S. homoeocarpa on potato dextrose agar (PDA) versus PDA amended with the discriminatory concentration of each fungicide. Field trials conducted at 3 locations in 2002 and 10 locations in 2003 revealed that the in vitro assays accurately predicted field efficacy for thiophanate-methyl. When used to screen 192 S. homoeocarpa isolates collected previously from 55 golf courses throughout Ohio, the in vitro assays revealed that 34 of the golf courses sampled had S. homoeocarpa resistant to thiophanate-methyl. S. homoeocarpa with reduced in vitro sensitivities was isolated from 18 and 1 golf courses for propiconazole and iprodione, respectively.


Botox as an adjuvant to tendon transfer for foot drop.
The authors' hypothesis was that weakening the gastrocnemius muscle at the time of tendon transfer will reduce the risk of rupture and facilitate faster rehabilitation with increased active dorsiflexion and improved overall outcome, because the transferred tendon is spared the antagonistic effect of the gastrocnemius during the early recovery period. A retrospective chart review identified 12 patients who underwent a tibialis posterior tendon (PTT) transfer with gastrocnemius Botox injection for foot drop. All statistical analyses were conducted using SAS version 9.2 (SAS Institute, Inc., Cary, North Carolina). There were no failures or tendon ruptures. All patients had zero dorsiflexion (DF) strength preoperatively, and those with DF strength documented postoperatively had a mean DF strength at final clinical evaluation of 3.9 (p < .001) with a mean active DF of 4°. It was concluded that gastrocnemius chemodenervation with Botox at the time of PTT transfer is a safe and effective means of restoring active dorsiflexion.


Role of the glutathione redox cycle in acquired and de novo multidrug resistance.
Drug resistance represents a major obstacle to successful cancer chemotherapy. However, the specific biochemical mechanisms responsible for clinical drug resistance are unknown. In these studies resistance to the antitumor agent adriamycin was found to involve two mechanisms, one that decreased drug accumulation by the P170 mechanism and another that altered the glutathione redox cycle, an important pathway in the detoxification of reactive oxygen. This dual mechanism of drug resistance was demonstrated in cell lines that had acquired the multidrug-resistant phenotype and in human colorectal cancer cells with de novo resistance. These studies support a model of acquired and de novo multidrug resistance that includes alterations in both drug accumulation and the glutathione redox cycle.


Adaptation of the Monetary Choice Questionnaire to accommodate extreme monetary discounting in cocaine users.
Delay discounting, which refers to the phenomenon that rewards decrease in subjective value as the delay associated with their receipt increases, is a paradigm that has been used extensively in substance abuse research to understand impulsive decision making. One common measure to assess delay discounting is the Monetary Choice Questionnaire (MCQ) developed by Kirby, Petry, and Bickel (1999). While the MCQ has great utility because of its simplicity and brief administration time, it is possible that the MCQ produces a ceiling effect in estimating delay discounting parameters in highly impulsive individuals. In the present study, we adapted the MCQ to attempt to address this ceiling effect by extending the original scale with 9 items, and we then compared scores on the original MCQ with the extended MCQ in a sample of active cocaine users. The ceiling effect, while observed in the original MCQ scores for over a quarter of the sample, was largely eliminated with the extended scale. Highly impulsive participants, whose scores on the extended scale exceeded the highest possible score on the original scale, had higher levels of sensation seeking compared to other participants, but not trait impulsivity. The extended MCQ may be useful in populations with high rates of impulsivity, where the original measure may underestimate discounting rates due to a ceiling effect.


Introduction of Glyoxylate Bypass Increases Hydrogen Gas Yield from Acetate and l-Glutamate in Rhodobacter sphaeroides.
Rhodobacter sphaeroides produces hydrogen gas (H2) from organic compounds via nitrogenase under anaerobic-light conditions in the presence of poor nitrogen sources, such as l-glutamate. R. sphaeroides utilizes the ethylmalonyl-coenzyme A (EMC) pathway for acetate assimilation, but its H2 yield from acetate in the presence of l-glutamate has been reported to be low. In this study, the deletion of ccr encoding crotonyl-coenzyme A (crotonyl-CoA) carboxylase/reductase, a key enzyme for the EMC pathway in R. sphaeroides, revealed that the EMC pathway is essential for H2 production from acetate and l-glutamate but not for growth and acetate consumption in the presence of l-glutamate. We introduced a plasmid expressing aceBA from Rhodobacter capsulatus encoding two key enzymes for the glyoxylate bypass into R. sphaeroides, which resulted in a 64% increase in H2 production. However, compared with the wild-type strain expressing heterologous aceBA genes, the strain with aceBA introduced in the genetic background of an EMC pathway-disrupted mutant showed a lower H2 yield. These results indicate that a combination of the endogenous EMC pathway and a heterologously expressed glyoxylate bypass is beneficial for H2 production. In addition, introduction of the glyoxylate bypass into a polyhydroxybutyrate (PHB) biosynthesis-disrupted mutant resulted in a delay in growth along with H2 production, although its H2 yield was comparable to that of the wild-type strain expressing heterologous aceBA genes. These results suggest that PHB production is important for fitness to the culture during H2 production from acetate and l-glutamate when both acetate-assimilating pathways are present.IMPORTANCE As an alternative to fossil fuel, H2 is a promising renewable energy source. Although photofermentative H2 production from acetate is key to developing an efficient process of biohydrogen production from biomass-derived sugars, H2 yields from acetate and l-glutamate by R. sphaeroides have been reported to be low. In this study, we observed that in addition to the endogenous EMC pathway, heterologous expression of the glyoxylate bypass in R. sphaeroides markedly increased H2 yields from acetate and l-glutamate. Therefore, this study provides a novel strategy for improving H2 yields from acetate in the presence of l-glutamate and contributes to a clear understanding of acetate metabolism in R. sphaeroides during photofermentative H2 production.


Esophageal saturation during antegrade cerebral perfusion: a preliminary report using visible light spectroscopy.
Visible light spectroscopy (VLS) is newer technology that measures real-time tissue oxygenation. It has been validated in detecting mucosal ischemia in adults. During complex neonatal heart surgery, antegrade cerebral perfusion (ACP) maintains cerebral saturation. Whether ACP maintains peripheral tissue perfusion in humans is not known. Five patients undergoing neonatal open heart surgery with hypothermic cardiopulmonary bypass (CPB) were studied using a VLS esophageal probe in addition to bilateral near infrared cerebral oximetry. Three of five patients required ACP for arch repair, while two patients did not. VLS and cerebral saturation data were collected and analyzed in 5 min intervals prior to CPB, during CPB, and during ACP. In the two patients undergoing heart surgery with routine hypothermic CPB, both cerebral and esophageal saturations were maintained. However in all three neonates requiring ACP, although cerebral saturations did not decrease, esophageal saturation fell below the ischemic threshold (35%). Following establishment of normal CPB, esophageal saturation returned to baseline. Antegrade cerebral perfusion maintains cerebral oxygen delivery, however, it does not adequately perfuse the esophagus in neonates. This could have clinical implications.


TRIM32, but not its muscular dystrophy-associated mutant, positively regulates and is targeted to autophagic degradation by p62/SQSTM1.
The tripartite motif (TRIM) proteins constitute a family of ubiquitin E3 ligases involved in a multitude of cellular processes, including protein homeostasis and autophagy. TRIM32 is characterized by six protein-protein interaction domains termed NHL, various point mutations in which are associated with limb-girdle-muscular dystrophy 2H (LGMD2H). Here, we show that TRIM32 is an autophagy substrate. Lysosomal degradation of TRIM32 was dependent on ATG7 and blocked by knockout of the five autophagy receptors p62 (also known as SQSTM1), NBR1, NDP52 (also known as CALCOCO2), TAX1BP1 and OPTN, pointing towards degradation by selective autophagy. p62 directed TRIM32 to lysosomal degradation, while TRIM32 mono-ubiquitylated p62 on lysine residues involved in regulation of p62 activity. Loss of TRIM32 impaired p62 sequestration, while reintroduction of TRIM32 facilitated p62 dot formation and its autophagic degradation. A TRIM32LGMD2H disease mutant was unable to undergo autophagic degradation and to mono-ubiquitylate p62, and its reintroduction into the TRIM32-knockout cells did not affect p62 dot formation. In light of the important roles of autophagy and p62 in muscle cell proteostasis, our results point towards impaired TRIM32-mediated regulation of p62 activity as a pathological mechanisms in LGMD2H.


The lateral patellar retinaculum defect: anatomical study using ultrasound.
The purpose of this study was to evaluate whether ultrasound allows precise assessment of a focal defect at the lateral patellar retinaculum (LPR) and to determine its anatomical significance. This work was initially undertaken in four cadaveric specimens and followed by high-resolution ultrasound study in 48 healthy adult volunteers (96 knees) by two musculoskeletal radiologists. Dimension and location of the LPR defect and its relations to adjacent anatomical structures were analyzed. A focal defect of the LPR through which vessels penetrated was constantly identified by ultrasound in our population. The mean transverse diameter of the defect was 2.4 mm (range, 1-6 mm). The defect was located a mean of 23.7 mm (range, 18-30 mm) proximal to the lateral tibial plateau and 6.6 mm (range, 4-9 mm) lateral to the patellar tendon. Anatomical dissection demonstrated that the LPR defect is related to perforating vessels that originate from the peripatellar anastomotic ring. The focal defect of the LPR can be depicted by ultrasonography. Reproducible anatomical landmarks for its detection could be defined. Our data suggest the LPR defect may be considered a normal variant rather than a pathologic change.


Inactivating and noninactivating Ca(2+)- and voltage-dependent K+ current in rat adrenal chromaffin cells.
The properties of Ca(2+)- and voltage-dependent K+ currents and their role in defining membrane potential were studied in cultured rat chromaffin cells. Two variants of large-conductance, Ca2+ and voltage-dependent BK channels, one noninactivating and one inactivating, were largely segregated among patches. Whole-cell noninactivating and inactivating currents resulting from each of these channels were segregated among different chromaffin cells. Cell-to-cell variation in the rate and extent of whole-cell current decay was not explained by differences in cytosolic [Ca2+] regulation among cells; rather, variation was due to differences in the intrinsic properties of the underlying BK channels. About 75% of rat chromaffin cells and patches express inactivating BK current (termed BKi) while the remainder express noninactivating BK current (termed BKs). The activation time course of both currents is similar, as is the dependence of activation on [Ca2+] and membrane potential. However, deactivation of BKi channels is slower than that of BKs channels. The functional role of these BK channel variants was studied in current-clamp recordings. Although both BKi and BKs currents contribute to action potential repolarization, cells expressing BKi current are better able to fire repetitively in response to constant current injection. Blockade of BKi current by charybdotoxin abolishes this behavior, showing that afterhyperpolarizations mediated by BKi current are permissive for repetitive firing. Thus, important properties of chromaffin cell membrane excitability are determined by the type of BK current expressed.


Reduced and Compressed Cisplatin-Based Chemotherapy in Children and Adolescents With Intermediate-Risk Extracranial Malignant Germ Cell Tumors: A Report From the Children's Oncology Group.
Purpose To investigate whether event-free survival (EFS) can be maintained among children and adolescents with intermediate-risk (IR) malignant germ cell tumors (MGCT) if the administration of cisplatin, etoposide, and bleomycin (PEb) is reduced from four to three cycles and compressed from 5 to 3 days per cycle. Patients and Methods In a phase 3, single-arm trial, patients with IR MGCT (stage II-IV testicular, II-III ovarian, I-II extragonadal, or stage I gonadal tumors with subsequent recurrence) received three cycles of PEb. A parametric comparator model specified that the observed EFS rate should not be significantly < 92%. As recommended for trials that test a reduction of therapy, a one-sided P value ≤ .10 was used to indicate statistical significance. In a post hoc analysis, we also compared results to the EFS rate of comparable patients treated with four cycles of PEb in two prior studies. Results Among 210 eligible patients enrolled from 2003 to 2011, 4-year EFS (EFS4) rate was 89% (95% confidence interval, 83% to 92%), which was significantly lower than the 92% threshold of the comparison model ( P = .08). Among 181 newly diagnosed patients, the EFS4 rate was 87%, compared with 92% for 92 comparable children in the historical cohort ( P = .15). The EFS4 rate was significantly associated with stage (stage I, 100%; stage II, 92%; stage III, 85%; and stage IV, 54%; P < .001). Conclusion The EFS rate for children with IR MGCT observed after three cycles of PEb was less than that of a prespecified parametric model, particularly for patients with higher-stage tumors. These data do not support a reduction in the number of cycles of PEb from four to three. However, further investigation of a reduction in the number of cycles for patients with lower-stage tumors is warranted.


Identification of PATCHED mutations in medulloblastomas by direct sequencing.
Medulloblastoma is the most common malignant embryonic tumors of the central nervous system. The nevoid basal cell carcinoma syndrome (NBCCS), which is caused by mutations of PTCH gene on chromosome 9q22, accounts for about 2% of all medulloblastomas. Previous studies of PTCH in sporadic medulloblastomas using single strand conformational polymorphism (SSCP) detected mutations in about 10% of the tumors. In this study, we directly sequenced the PTCH gene in 20 sporadic medulloblastoma DNA samples. A nonsense mutation (Q694X) and a splice site alteration (2875+1G>A) were identified in two of the samples. The mutations are predicted to result in a truncated PTCH protein and aberrant splicing, respectively. In both cases, only the mutant alleles were identified, indicating that the mutations were associated with loss of the wild-type PTCH allele in the tumor cells. Several novel variants, including 1653T>C, 1672C>T, and 2292C>T, were also found in these tumor samples. One of the two mutations detected in this study had been missed by SSCP, suggesting that the true rate of PTCH mutations in sporadic medulloblastomas may be underestimated by SSCP screening. Nevertheless, the frequency of mutations in this study did not differ from previous reports.


Site-Specific Neuromodulation of Detrusor and External Urethral Sphincter by Epidural Spinal Cord Stimulation.
Impairments of the lower urinary tract function including urine storage and voiding are widely spread among patients with spinal cord injuries. The management of such patients includes bladder catheterization, surgical and pharmacological approaches, which reduce the morbidity from urinary tract-related complications. However, to date, there is no effective treatment of neurogenic bladder and restoration of urinary function. In the present study, we examined neuromodulation of detrusor (Detr) and external urethral sphincter by epidural electrical stimulation (EES) of lumbar and sacral regions of the spinal cord in chronic rats. To our knowledge, it is the first chronic study where detrusor and external urethral sphincter signals were recorded simultaneously to monitor their neuromodulation by site-specific spinal cord stimulation (SCS). The data obtained demonstrate that activation of detrusor muscle mainly occurs during the stimulation of the upper lumbar (L1) and lower lumbar (L5-L6) spinal segments whereas external urethral sphincter was activated predominantly by sacral stimulation. These findings can be used for the development of neurorehabilitation strategies based on spinal cord epidural stimulation for autonomic function recovery after severe spinal cord injury (SCI).


Sociodemographic Disparities in the Receipt of Adjuvant Chemotherapy Among Patients With Resected Stage I-III Pancreatic Adenocarcinoma.
Adjuvant therapy for resected pancreatic adenocarcinoma was given a category 1 NCCN recommendation in 2000, yet many patients do not receive chemotherapy after definitive surgery. Whether sociodemographic disparities exist for receipt of adjuvant chemotherapy is poorly understood. The National Cancer Database was used to identify patients diagnosed with nonmetastatic pancreatic adenocarcinoma who underwent definitive surgery from 2004 through 2015. Multivariable logistic regression defined the adjusted odds ratio (aOR) and associated 95% CI of receipt of adjuvant chemotherapy. Among patients receiving chemotherapy, multivariable logistic regression assessed the odds of treatment with multiagent chemotherapy. Among 18,463 patients, 11,288 (61.1%) received any adjuvant chemotherapy. Sociodemographic factors inversely associated with receipt of any adjuvant chemotherapy included uninsured status (aOR, 0.61; 95% CI, 0.50-0.74), Medicaid insurance (aOR, 0.66; 95% CI, 0.57-0.77), and lower income (P<.001 for all income levels compared with ≥$46,000). Black race (aOR, 0.72; 95% CI, 0.57-0.90) and female sex (aOR, 0.75; 95% CI, 0.65-0.86) were associated with lower odds of receiving multiagent chemotherapy. There was a statistically significant interaction term between black race and age/comorbidity status (P=.03), such that 26.4% of black versus 35.8% of nonblack young (aged ≤65 years) and healthy (Charlson-Deyo comorbidity score 0) patients received multiagent adjuvant chemotherapy (P=.006), whereas multiagent adjuvant chemotherapy rates were similar among patients who were not young and healthy (P=.15). In this nationally representative study, receipt of adjuvant chemotherapy appeared to be associated with sociodemographic characteristics, independent of clinical factors. Sociodemographic differences in receipt of adjuvant chemotherapy may represent a missed opportunity for improving outcomes and a driver of oncologic disparities.


Modulation of endotoxicity of Shigella generalized modules for membrane antigens (GMMA) by genetic lipid A modifications: relative activation of TLR4 and TLR2 pathways in different mutants.
Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from Shigella. As GMMA are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (LPS). We examined ways of reducing their reactogenicity by modifying lipid A, the endotoxic part of LPS, through deletion of late acyltransferase genes, msbB or htrB, in GMMA-producing Shigella sonnei and Shigella flexneri strains. GMMA with resulting penta-acylated lipid A from the msbB mutants showed a 600-fold reduced ability, and GMMA from the S. sonnei ΔhtrB mutant showed a 60,000-fold reduced ability compared with GMMA with wild-type lipid A to stimulate human Toll-like receptor 4 (TLR4) in a reporter cell line. In human peripheral blood mononuclear cells, GMMA with penta-acylated lipid A showed a marked reduction in induction of inflammatory cytokines (S. sonnei ΔhtrB, 800-fold; ΔmsbB mutants, 300-fold). We found that the residual activity of these GMMA is largely due to non-lipid A-related TLR2 activation. In contrast, in the S. flexneri ΔhtrB mutant, a compensatory lipid A palmitoleoylation resulted in GMMA with hexa-acylated lipid A with ∼10-fold higher activity to stimulate peripheral blood mononuclear cells than GMMA with penta-acylated lipid A, mostly due to retained TLR4 activity. Thus, for use as vaccines, GMMA will likely require lipid A penta-acylation. The results identify the relative contributions of TLR4 and TLR2 activation by GMMA, which need to be taken into consideration for GMMA vaccine development.


Local field potential modeling predicts dense activation in cerebellar granule cells clusters under LTP and LTD control.
Local field-potentials (LFPs) are generated by neuronal ensembles and contain information about the activity of single neurons. Here, the LFPs of the cerebellar granular layer and their changes during long-term synaptic plasticity (LTP and LTD) were recorded in response to punctate facial stimulation in the rat in vivo. The LFP comprised a trigeminal (T) and a cortical (C) wave. T and C, which derived from independent granule cell clusters, co-varied during LTP and LTD. To extract information about the underlying cellular activities, the LFP was reconstructed using a repetitive convolution (ReConv) of the extracellular potential generated by a detailed multicompartmental model of the granule cell. The mossy fiber input patterns were determined using a Blind Source Separation (BSS) algorithm. The major component of the LFP was generated by the granule cell spike Na(+) current, which caused a powerful sink in the axon initial segment with the source located in the soma and dendrites. Reproducing the LFP changes observed during LTP and LTD required modifications in both release probability and intrinsic excitability at the mossy fiber-granule cells relay. Synaptic plasticity and Golgi cell feed-forward inhibition proved critical for controlling the percentage of active granule cells, which was 11% in standard conditions but ranged from 3% during LTD to 21% during LTP and raised over 50% when inhibition was reduced. The emerging picture is that of independent (but neighboring) trigeminal and cortical channels, in which synaptic plasticity and feed-forward inhibition effectively regulate the number of discharging granule cells and emitted spikes generating "dense" activity clusters in the cerebellar granular layer.


Nursing care for patients with COVID-19 on extracorporeal membrane oxygenation (ECMO) support.
In Japan, four medical facilities including our own - the National Center for Global health and Medicine (NCGM) - have been designated for the treatment of specified infectious diseases by the Minister of Health, Labour, and Welfare. Here, we report our nursing care for patients with severe COVID-19 on extracorporeal membrane oxygenation (ECMO) support. In addition to infection control measures in the form of an N95 mask, a water-repellent isolation gown, a cap, a shielded mask on top of the N95, and double-layered gloves, nurses were required to wear one-piece suits (DuPont™ Tyvek®) and use powered air-purifying respirators (PAPRs). While closed system catheters are normally changed once a day to limit aerosol exposure, they are now changed once every 4 days. Nursing care included equipment checks, monitoring of hemodynamics and respiratory status, management of anticoagulants, observation of the patients general condition, management of sedatives and analgesics, prevention of medical device-related pressure ulcers and bedsores, and maintenance of hygiene. Fundamentally sound nursing remains the best practice for patient treatment and management. During nursing care for patients with COVID-19 on ECMO, infection control measures should be faithfully and properly followed.


Trace but not delay fear conditioning requires attention and the anterior cingulate cortex.
Higher cognitive functions such as attention have been difficult to model in genetically tractable organisms. In humans, attention-distracting stimuli interfere with trace but not delay conditioning, two forms of associative learning. Attention has also been correlated with activation of anterior cingulate cortex (ACC), but its functional significance is unclear. Here we show that a visual distractor interferes selectively with trace but not delay auditory fear conditioning in mice. Trace conditioning is associated with increased neuronal activity in ACC, as assayed by relative levels of c-fos expression, and is selectively impaired by lesions of this structure. The effects of the ACC lesions are unlikely to be caused by indirect impairment of the hippocampus, which is required for mnemonic aspects of trace conditioning. These data suggest that trace conditioning may be useful for studying neural substrates of attention in mice, and implicate the ACC as one such substrate.


The Quality of Online Resources Available to Patients Regarding Cannabidiol for Symptomatic Relief of Hip or Knee Arthritis is Poor.
This study aimed to assess the quality of online resources pertaining to cannabidiol (CBD) for the nonoperative management of hip and knee arthritis. Websites were identified on the three most popular global search engines using terms relevant to CBD, hip or knee pain, and arthritis. Websites were scored based on a 25-point scale regarding diagnosis, evaluation, and treatment of hip and knee pathologies. The initial search yielded 287 results, and 94 websites were analyzed after meeting inclusion criteria. The average Flesch-Kincaid reading level was 48, corresponding to a college education level. Mean website score was poor at 7.46 (SD 3.51) of 25 (29.8%). Websites published by physicians had statistically higher scores (P = 0.03). Many online resources regarding CBD use for hip and knee arthritis are available; however, the readability is more advanced than recommended by the National Institutes of Health. Very few resources are sponsored by physicians or professional organizations, and many are overtly sales oriented. Patients should be counseled that the information available online on this topic is generally unreliable. Surgeons and professional health organizations should play a stronger role in providing balanced resources to patients regarding CBD use for hip and knee arthritis.


Women's experiences of pregnancy, childbirth, and the postnatal period in The Gambia: a qualitative study.
In sub-Saharan African countries, there are unique cultural factors and adverse physical conditions that contribute to women's experiences of pregnancy and birth. The objective of this study was to qualitatively explore women's experiences of pregnancy, childbirth, the postnatal period, and maternal psychological distress in The Gambia. Semi-structured interviews were carried out with 55 women who had given birth within the previous year. Thematic analysis identified five themes: (1) transition to adulthood, (2) physical difficulties, (3) value of children in relation to others, (4) children as a strain, and (5) going through it alone. The results suggest that having a child is a defining point in women's lives associated with happiness and joy. However, women also described situations which could lead to unhappiness and distress in the perinatal period. A child conceived out of wedlock or a baby girl can be sources of distress because of negative cultural perceptions. The strain of having a child, particularly the additional financial burden, and minimal support from men were also a concern for women. Finally, women recognized the danger associated with delivery and expressed recurrent worries of complications during childbirth which could result in the death of them or the baby. Further research is needed to identify women vulnerable to psychological distress so that health services and target interventions can be developed accordingly.


Computer simulation models are implementable as replacements for animal experiments.
It has become increasingly difficult to perform animal experiments, because of issues related to the procurement of animals, and strict regulations and ethical issues related to their use. As a result, it is felt that the teaching of pharmacology should be more clinically oriented and that unnecessary animal experimentation should be avoided. Although a number of computer simulation models (CSMs) are available, they are not being widely used. Interactive demonstrations were conducted to encourage the departmental faculty to use CSMs. Four different animal experiments were selected, that dealt with actions of autonomic drugs. The students observed demonstrations of animal experiments involving conventional methods and the use of CSMs. This was followed by hands-on experience of the same experiment, but using CSMs in small groups, instead of hands-on experience with the animal procedures. Test scores and feedback showed that there was better understanding of the mechanisms of action of the drugs, gained in a shorter time. The majority of the students found the teaching programme used to be good to excellent. CSMs can be used repeatedly and independently by students, and this avoids unnecessary experimentation and also causing pain and trauma to animals. The CSM programme can be implemented in existing teaching schedules for pharmacology undergraduate teaching with basic infrastructure support, and is readily adaptable for use by other institutes.


High Ultraviolet Absorption in Colloidal Gallium Nanoparticles Prepared from Thermal Evaporation.
New methods for the production of colloidal Ga nanoparticles (GaNPs) are introduced based on the evaporation of gallium on expendable aluminum zinc oxide (AZO) layer. The nanoparticles can be prepared in aqueous or organic solvents such as tetrahydrofuran in order to be used in different sensing applications. The particles had a quasi mono-modal distribution with diameters ranging from 10 nm to 80 nm, and their aggregation status depended on the solvent nature. Compared to common chemical synthesis, our method assures higher yield with the possibility of tailoring particles size by adjusting the deposition time. The GaNPs have been studied by spectrophotometry to obtain the absorption spectra. The colloidal solutions exhibit strong plasmonic absorption in the ultra violet (UV) region around 280 nm, whose width and intensity mainly depend on the nanoparticles dimensions and their aggregation state. With regard to the colloidal GaNPs flocculate behavior, the water solvent case has been investigated for different pH values, showing UV-visible absorption because of the formation of NPs clusters. Using discrete dipole approximation (DDA) method simulations, a close connection between the UV absorption and NPs with a diameter smaller than ~40 nm was observed.


Monocular optical constraints on collision control.
A simulated ball-hitting task was used to explore the optical basis for collision control. Ball speed and size were manipulated in Experiments 1 and 2. Results showed a tendency for participants to respond earlier to slower and larger balls. Early in practice, participants would consistently miss the slowest and largest balls. Experiments 3 and 4 examined performance as a function of the range of speeds. Performance for identical speeds differed depending on whether the speeds were fastest or slowest within a range. Asymmetric transfer between the 2 ranges of speeds showed that those trained with slow speeds were very successful when tested with a faster range of speeds. Those trained with fast speeds did not do as well when tested on slower speeds. The pattern of results across 4 experiments suggests that participants were using optical angle and expansion rate as separate degrees of freedom for solving the collision task.


Effectiveness of a Hospital-Based Multidisciplinary Pediatric Weight Management Program: Two-Year Outcomes of PHIT Kids.
For children with obesity, long-term sustainability of weight loss after treatment is difficult to achieve. This study examined 2-year anthropometric outcomes of a moderately intensive group behaviorally based weight management program. One hundred seventy-three children with obesity ages 8-18 years participated with their parent or adult caregiver in a 24-week multicomponent intervention, which was followed by monthly sessions for a total of 2 years. Children were considered treatment completers if they attended ≥50% of the 24 weekly sessions. A multilevel model (multiple assessment time points nested within participants) was used to test person-level change in BMI z-score (BMIz) for program completers between (1) pre- and post-treatment, (2) pretreatment and 24-month follow-up, (3) post-treatment and 12-month follow-up, and (4) post-treatment and 24-month follow-up. One hundred twenty-four (72%) of the participants completed the 24-week intervention. Significant reductions in BMIz were observed over the course of treatment (β = -0.03; standard error [SE] = 0.004; t = -6.85; p < 0.001). Completers showed a significant reduction in BMIz between initiation of treatment and 2-year follow-up (n = 110 at 24 weeks; n = 38 at 24 months; β = -0.02; SE = 0.005; t = -4.12; p < 0.001). Children did not show any significant changes in BMIz between post-treatment and 24-month follow-up (β = -0.006; SE = 0.011; t = -0.61; p = 0.54), suggesting that treatment effects were maintained. Children maintained treatment gains achieved during a 24-week family-based behavioral weight management program at 2-year follow-up. Although these findings suggest that gains are sustainable, further research is needed to understand how these long-term changes impact child health.


Use of transcutaneous electrical nerve stimulation as an adjunctive to epidural analgesia in the management of acute thoracotomy pain.
The present randomized study was conducted in our institute of pulmonary medicine and tuberculosis over a period of 1 year. This study aimed to evaluate the effectiveness of transcutaneous electrical nerve stimulation (TENS) as an adjunctive to thoracic epidural analgesia for the treatment of postoperative pain in patients who underwent posterolateral thoracotomy for decortication of lung. Sixty patients in the age group 15-40 years scheduled to undergo elective posterolateral thoracotomy were divided into two groups of 30 each. Patients were alternatively assigned to one of the groups. In group I, only thoracic epidural analgesia with local anaesthetics was given at regular intervals; however, an identical apparatus which did not deliver an electric current was applied to the control (i.e. group I) patients. While in group II, TENS was started immediately in the recovery period in addition to the epidural analgesia. A 0-10 visual analog scale (VAS) was used to assess pain at regular intervals. The haemodynamics were also studied at regular intervals of 2 h for the first 10 h after the surgery. When the VAS score was more than three, intramuscular analgesia with diclofenac sodium was given. The VAS score and the systolic blood pressure were comparable in the immediate postoperative period (P = NS) but the VAS score was significantly less in group II at 2, 4, 6, 8 h (P < 0.01, P < 0.05, P < 0.05, P < 0.05, respectively), and at 10 h the P value was not significant. Similarly, the systolic blood pressure was significantly less in group II at 2, 4, 6 h after surgery, that is P < 0.02, P < 0.01, P < 0.01, respectively, but at 8 and 10 h the pressures were comparable in both the groups. Adding TENS to epidural analgesia led to a significant reduction in pain with no sequelae. The haemodynamics were significantly stable in group II compared to group I. TENS is a valuable strategy to alleviate postoperative pain following thoracic surgery with no side effects and with a good haemodynamic stability; however, the effects are short lasting.


Resource availability and competition shape the evolution of survival and growth ability in a bacterial community.
Resource availability is one of the main factors determining the ecological dynamics of populations or species. Fluctuations in resource availability can increase or decrease the intensity of resource competition. Resource availability and competition can also cause evolutionary changes in life-history traits. We studied how community structure and resource fluctuations affect the evolution of fitness related traits using a two-species bacterial model system. Replicated populations of Serratia marcescens (copiotroph) and Novosphingobium capsulatum (oligotroph) were reared alone or together in environments with intergenerational, pulsed resource renewal. The comparison of ancestral and evolved bacterial clones with 1 or 13 weeks history in pulsed resource environment revealed species-specific changes in life-history traits. Co-evolution with S. marcescens caused N. capsulatum clones to grow faster. The evolved S. marcescens clones had higher survival and slower growth rate then their ancestor. The survival increased in all treatments after one week, and thereafter continued to increase only in the S. marcescens monocultures that experienced large resource pulses. Though adaptive radiation is often reported in evolution studies with bacteria, clonal variation increased only in N. capsulatum growth rate. Our results suggest that S. marcescens adapted to the resource renewal cycle whereas N. capsulatum was more affected by the interspecific competition. Our results exemplify species-specific evolutionary response to both competition and environmental variation.


Dual inhibition of the PI3K/mTOR pathway increases tumor radiosensitivity by normalizing tumor vasculature.
The aberrant vascular architecture of solid tumors results in hypoxia that limits the efficacy of radiotherapy. Vascular normalization using antiangiogenic agents has been proposed as a means to improve radiation therapy by enhancing tumor oxygenation, but only short-lived effects for this strategy have been reported so far. Here, we show that NVP-BEZ235, a dual inhibitor of phosphoinositide-3-kinase (PI3K) and mTOR, can improve tumor oxygenation and vascular structure over a prolonged period that achieves the aim of effective vascular normalization. Because PI3K inhibition can radiosensitize tumor cells themselves, our experimental design explicitly distinguished effects on the blood vasculature versus tumor cells. Drug administration coincident with radiation enhanced the delay in tumor growth without changing tumor oxygenation, establishing that radiosensitization is a component of the response. However, the enhanced growth delay was substantially greater after induction of vascular normalization, meaning that this treatment enhanced the tumoral radioresponse. Importantly, changes in vascular morphology persisted throughout the entire course of the experiment. Our findings indicated that targeting the PI3K/mTOR pathway can modulate the tumor microenvironment to induce a prolonged normalization of blood vessels. The substantial therapeutic gain observed after combination of NVP-BEZ235 with irradiation has conceptual implications for cancer therapy and could be of broad translational importance.


Nox2 contributes to age-related oxidative damage to neurons and the cerebral vasculature.
Oxidative stress plays an important role in aging-related neurodegeneration. This study used littermates of WT and Nox2-knockout (Nox2KO) mice plus endothelial cell-specific human Nox2 overexpression-transgenic (HuNox2Tg) mice to investigate Nox2-derived ROS in brain aging. Compared with young WT mice (3-4 months), aging WT mice (20-22 months) had obvious metabolic disorders and loss of locomotor activity. Aging WT brains had high levels of angiotensin II (Ang II) and ROS production; activation of ERK1/2, p53, and γH2AX; and losses of capillaries and neurons. However, these abnormalities were markedly reduced in aging Nox2KO brains. HuNox2Tg brains at middle age (11-12 months) already had high levels of ROS production and activation of stress signaling pathways similar to those found in aging WT brains. The mechanism of Ang II-induced endothelial Nox2 activation in capillary damage was examined using primary brain microvascular endothelial cells. The clinical significance of Nox2-derived ROS in aging-related loss of cerebral capillaries and neurons was investigated using postmortem midbrain tissues of young (25-38 years) and elderly (61-85 years) adults. In conclusion, Nox2 activation is an important mechanism in aging-related cerebral capillary rarefaction and reduced brain function, with the possibility of a key role for endothelial cells.


Human intestinal tissue-resident memory T cells comprise transcriptionally and functionally distinct subsets.
Tissue-resident memory T (TRM) cells provide key adaptive immune responses in infection, cancer, and autoimmunity. However, transcriptional heterogeneity of human intestinal TRM cells remains undefined. Here, we investigate transcriptional and functional heterogeneity of human TRM cells through study of donor-derived TRM cells from intestinal transplant recipients. Single-cell transcriptional profiling identifies two transcriptional states of CD8+ TRM cells, delineated by ITGAE and ITGB2 expression. We define a transcriptional signature discriminating these populations, including differential expression of cytotoxicity- and residency-associated genes. Flow cytometry of recipient-derived cells infiltrating the graft, and lymphocytes from healthy gut, confirm these CD8+ TRM phenotypes. CD8+ CD69+CD103+ TRM cells produce interleukin-2 (IL-2) and demonstrate greater polyfunctional cytokine production, whereas β2-integrin+CD69+CD103- TRM cells have higher granzyme expression. Analysis of intestinal CD4+ T cells identifies several parallels, including a β2-integrin+ population. Together, these results describe the transcriptional, phenotypic, and functional heterogeneity of human intestinal CD4+ and CD8+ TRM cells.


The effect of drop volume and micropillar shape on the apparent contact angle of ordered microstructured surfaces.
In the present paper, we propose a new theoretical approach to evaluate the shape and apparent contact angle (ACA) of a drop gently deposited on microstructured superhydrophobic surfaces. We exploit the very large separation of scales between the drop size and the features of the micromorphology of the interface to propose a numerical methodology to calculate the apparent contact area and apparent contact angle. In agreement with very recent experiments, calculations show that, in the case of surfaces made of conical micropillars, the ACA may take values very close to 180° not depending on the size of the liquid drop. At large drop volumes, the shape of the drop deviates from the spherical one as a result of the gravity effects, but it is noteworthy that the apparent contact angle does not change at all. Our calculations shows that this holds true also for different pillar shapes, showing that, for any given Young contact angle of the solid constituting the pillars, the ACA is an intrinsic property of the surface microgeometry.


The Human Phenotype Ontology in 2021.
The Human Phenotype Ontology (HPO, https://hpo.jax.org) was launched in 2008 to provide a comprehensive logical standard to describe and computationally analyze phenotypic abnormalities found in human disease. The HPO is now a worldwide standard for phenotype exchange. The HPO has grown steadily since its inception due to considerable contributions from clinical experts and researchers from a diverse range of disciplines. Here, we present recent major extensions of the HPO for neurology, nephrology, immunology, pulmonology, newborn screening, and other areas. For example, the seizure subontology now reflects the International League Against Epilepsy (ILAE) guidelines and these enhancements have already shown clinical validity. We present new efforts to harmonize computational definitions of phenotypic abnormalities across the HPO and multiple phenotype ontologies used for animal models of disease. These efforts will benefit software such as Exomiser by improving the accuracy and scope of cross-species phenotype matching. The computational modeling strategy used by the HPO to define disease entities and phenotypic features and distinguish between them is explained in detail.We also report on recent efforts to translate the HPO into indigenous languages. Finally, we summarize recent advances in the use of HPO in electronic health record systems.


Pituitary dysfunction after traumatic brain injury: a clinical and pathophysiological approach.
Traumatic brain injury (TBI) is a growing public health problem worldwide and is a leading cause of death and disability. The causes of TBI include motor vehicle accidents, which are the most common cause, falls, acts of violence, sports-related head traumas, and war accidents including blast-related brain injuries. Recently, pituitary dysfunction has also been described in boxers and kickboxers. Neuroendocrine dysfunction due to TBI was described for the first time in 1918. Only case reports and small case series were reported until 2000, but since then pituitary function in TBI victims has been investigated in more detail. The frequency of hypopituitarism after TBI varies widely among different studies (15-50% of the patients with TBI in most studies). The estimates of persistent hypopituitarism decrease to 12% if repeated testing is applied. GH is the most common hormone lost after TBI, followed by ACTH, gonadotropins (FSH and LH), and TSH. The underlying mechanisms responsible for pituitary dysfunction after TBI are not entirely clear; however, recent studies have shown that genetic predisposition and autoimmunity may have a role. Hypopituitarism after TBI may have a negative impact on the pace or degree of functional recovery and cognition. What is not clear is whether treatment of hypopituitarism has a beneficial effect on specific function. In this review, the current data related to anterior pituitary dysfunction after TBI in adult patients are updated, and guidelines for the diagnosis, follow-up strategies, and therapeutic approaches are reported.


Krüppel-like factor 6 is frequently down-regulated and induces apoptosis in non-small cell lung cancer cells.
Krüppel-like factor 6 (KLF6) is a ubiquitously expressed zinc finger transcriptional factor, which has been suggested to be a candidate tumor suppressor gene in prostate cancer and astrocytic glioma. Because KLF6 is located at chromosome 10p15, where non-small cell lung cancers (NSCLCs) also exhibit frequent allelic loss, we hypothesized that the inactivation of KLF6 is also involved in the development of NSCLC. To determine this, we performed mutational analysis for 105 NSCLCs, including 9 cell lines and 96 primary tumors, and Northern blot analysis for 74 NSCLCs, including the 9 cell lines and 65 primary tumors. Although somatic mutations were not detected in the coding sequence of KLF6, expression of KLF6 mRNA was down-regulated in the 9 cell lines and in 55 (85%) of the 65 primary tumors compared with normal lung tissue. Treatment of two cell lines expressing KLF6 at low levels with 5-azacytidine did not induce KLF6 expression, suggesting that KLF6 down-regulation is not due to promoter hypermethylation. We also performed loss of heterozygosity (LOH) analysis using the laser capture microdissection technique, and found that 21 of 62 (34%) informative samples had LOH in the KLF6 gene locus. Comparing the LOH status with mRNA expression of KLF6, we found that 14 of the 14 (100%) samples with LOH showed KLF6 down-regulation, and that even 23 of 31 (74%) samples without LOH also showed this down-regulation. We also studied the expression of the WAF1 gene, a possible downstream gene of KLF6, and detected simultaneous down-regulation of WAF1 and KLF6 mRNA in 6 of 9 (67%) cell lines and 48 of the 55 (87%) primary tumors, although there was not a significant association between loss of KLF6 and WAF1 expression. Furthermore, colony formation assay of two NSCLC cell lines (NCI-H1299 and NCI-H2009) induced a markedly reduced colony formation by KLF6 transfection, and Annexin V staining and terminal deoxynucleotidyl transferase-mediated nick end labeling assays revealed that KLF6 induced apoptosis. Our present studies demonstrated that KLF6 is frequently down-regulated in NSCLC and suppresses tumor growth via induction of apoptosis in NSCLC, which may suggest that KLF6 is a tumor suppressor for NSCLC.


Relative proliferative rates of limbal and corneal epithelia. Implications of corneal epithelial migration, circadian rhythm, and suprabasally located DNA-synthesizing keratinocytes.
An important element of the recently proposed limbal stem cell model is that corneal epithelial cells migrate centripetally. The driving force for this migration is unknown, although it has been suggested that limbal epithelium, proliferates at a higher rate than central corneal epithelium, thus creating a population pressure toward the central cornea. This hypothesis was tested by measuring the relative proliferative rates of limbal and central corneal epithelia using 3H-thymidine autoradiographic techniques. The results indicate that, in both the New Zealand white rabbit and SENCAR mouse, the labeling index (LI) of limbal epithelium is actually lower than that of central corneal epithelium. This difference in LI persists throughout the circadian rhythm cycle. These results suggest that population pressure per se cannot be responsible for the centripetal migration of corneal epithelium and raise the possibility that preferential desquamation of central corneal epithelium may "draw" peripheral cells toward the central cornea. In both epithelia, the LI peak precedes the mitotic index (MI) peak during circadian cycle by 4-6 hr. These data therefore are in close agreement with earlier results on several nonocular stratified epithelia but contradict an earlier suggestion that the LI and MI peaks of corneal epithelium coincide. Finally, although most of the 3H-thymidine incorporating cells in central cornea may appear to be suprabasally located, they are only partially displaced into the suprabasal compartment. In most cases, such cells are still connected with the basement membrane through a thin stalk of cytoplasm. Since corneal epithelium rests on an exceptionally flat and rigid substratum, an increase in cellular volume in DNA-synthesizing cells may not be tolerated well in an already crowded basal layer. This may explain why an unusually large proportion of DNA-synthesizing cells are expelled preferentially into either a "second tier basal layer" or into the suprabasal compartment.


Effect of refractive index mismatch on multi-photon direct laser writing.
This work reports how the process of three-dimensional multi-photon direct laser writing (mpDLW) is affected when there is a small mismatch in refractive index between the material being patterned and the medium in which the focusing objective is immersed. Suspended-line microstructures were fabricated by mpDLW in the cross-linkable epoxide SU-8 as a function of focus depth and average incident power. It is found that even a small refractive index contrast of Δn = + 0.08 causes significant variation in feature width and height throughout the depth of the material. In particular, both the width and height of features can either increase or decrease with depth, depending upon how much the average incident laser power exceeds the threshold for writing. Vectorial diffraction theory is used to obtain insight into the origin of the effect and how to compensate for it. We demonstrate that varying the average focused power is a practical means for controlling the variation in feature size with focal depth.


SMFM Consult Series #46: Evaluation and management of polyhydramnios.
Polyhydramnios, or hydramnios, is an abnormal increase in the volume of amniotic fluid. Identification of polyhydramnios should prompt a search for an underlying etiology. Although most cases of mild polyhydramnios are idiopathic, the 2 most common pathologic causes are maternal diabetes mellitus and fetal anomalies, some of which are associated with genetic syndromes. Other causes of polyhydramnios include congenital infection and alloimmunization. The purpose of this document is to provide guidance on the evaluation and management of polyhydramnios. The following are Society for Maternal-Fetal Medicine recommendations: (1) we suggest that polyhydramnios in singleton pregnancies be defined as either a deepest vertical pocket of ≥8 cm or an amniotic fluid index of ≥24 cm (GRADE 2C); (2) we recommend that amnioreduction be considered only for the indication of severe maternal discomfort, dyspnea, or both in the setting of severe polyhydramnios (GRADE 1C); (3) we recommend that indomethacin should not be used for the sole purpose of decreasing amniotic fluid in the setting of polyhydramnios (GRADE 1B); (4) we suggest that antenatal fetal surveillance is not required for the sole indication of mild idiopathic polyhydramnios (GRADE 2C); (5) we recommend that labor should be allowed to occur spontaneously at term for women with mild idiopathic polyhydramnios; that induction, if planned, should not occur at <39 weeks of gestation in the absence of other indications; and that mode of delivery should be determined based on usual obstetric indications (GRADE 1C); and (6) we recommend that women with severe polyhydramnios deliver at a tertiary center due to the significant possibility that fetal anomalies may be present (GRADE 1C).


The genome sequence of an H11N2 avian influenza virus from a Thick-billed Murre (Uria lomvia) shows marine-specific and regional patterns of relationships to other viruses.
Influenza A viruses infect a range of host species, including various mammals and more than 100 species of birds. For avian influenza viruses (AIV), prevalence varies between different groups of birds, with waterfowl showing the highest prevalence. We have sequenced the complete genome of A/Thick-billed Murre/Newfoundland/031/2007(H11N2), an AIV identified in the pelagic seabird, Thick-billed Murre (Uria lomvia). This represents the first complete genome sequence of an AIV from this host species, and only the second complete genome sequence from a seabird in the alcid group. All of the virus segments fall within the American avian lineage. Several of the segments show a close relationship to AIV identified in other marine host species, and also a strong geographic association with other AIV sequences from the northeastern coast of North America from recent years. The identification of this virus, and the growing number of AIV identified in seabird species, indicates these marine birds could be underappreciated host species. This has potential consequences for global influenza dynamics because of the seasonal distributions and migratory patterns of this group of birds.


Definition of transcriptional pause elements in fission yeast.
Downstream elements (DSEs) with transcriptional pausing activity play an important role in transcription termination of RNA polymerase II. We have defined two such DSEs in Schizosaccharomyces pombe, one for the ura4 gene and a new one in the 3'-end region of the nmt2 gene. Although these DSEs do not have sequence homology, both are orientation specific and are composed of multiple and redundant sequence elements that work together to achieve full pausing activity. Previous studies on the nmt1 and nmt2 genes revealed that transcription extends several kilobases past the genes' poly(A) sites. We show that the insertion of either DSE immediately downstream of the nmt1 poly(A) site induces more immediate termination. nmt2 termination efficiency can be increased by moving the DSE closer to the poly(A) site. These results suggest that DSEs may be a common feature in yeast genes.


Construct validation of a non-exercise measure of cardiorespiratory fitness in older adults.
Cardiorespiratory fitness (CRF) is associated with a decreased risk of all-cause mortality but is rarely assessed in medical settings due to burdens of time, cost, risk, and resources. The purpose of this study was to test the construct validity of a regression equation developed by Jurca and colleagues (2005) to estimate CRF without exercise testing in community dwelling older adults. Participants (n = 172) aged 60 to 80 years with no contraindications to submaximal or maximal exercise testing completed a maximal graded exercise test (GXT) and the submaximal Rockport 1-mile walk test on separate occasions. Data included in the regression equation (age, sex, body mass index, resting heart rate, and physical activity) were obtained via measurement or self-report. Participants also reported presence of cardiovascular conditions. The multiple R for the regression equation was .72, p < .001 and CRF estimated from this equation was significantly correlated with the MET value from the GXT (r = 0.66) and with CRF estimated from submaximal field testing (r = 0.67). All three CRF indices were significantly and inversely associated with reporting more cardiovascular conditions. This research provides preliminary evidence that a non-exercise estimate of CRF is at least as valid as field test estimates of CRF and represents a low-risk, low-cost, and expedient method for estimating fitness in older adults.


A common 5'-UTR variant in MATE2-K is associated with poor response to metformin.
Multidrug and toxin extrusion 2 (MATE2-K (SLC47A2)), a polyspecific organic cation exporter, facilitates the renal elimination of the antidiabetes drug metformin. In this study, we characterized genetic variants of MATE2-K, determined their association with metformin response, and elucidated their impact by means of a comparative protein structure model. Four nonsynonymous variants and four variants in the MATE2-K basal promoter region were identified from ethnically diverse populations. Two nonsynonymous variants-c.485C>T and c.1177G>A-were shown to be associated with significantly lower metformin uptake and reduction in protein expression levels. MATE2-K basal promoter haplotypes containing the most common variant, g.-130G>A (>26% allele frequency), were associated with a significant increase in luciferase activities and reduced binding to the transcriptional repressor myeloid zinc finger 1 (MZF-1). Patients with diabetes who were homozygous for g.-130A had a significantly poorer response to metformin treatment, assessed as relative change in glycated hemoglobin (HbA1c) (-0.027 (-0.076, 0.033)), as compared with carriers of the reference allele, g.-130G (-0.15 (-0.17, -0.13)) (P=0.002). Our study showed that MATE2-K plays a role in the antidiabetes response to metformin.


Recommendations for translation and reliability testing of International Spinal Cord Injury Data Sets.
To provide recommendations regarding translation and reliability testing of International Spinal Cord Injury (SCI) Data Sets. The Executive Committee for the International SCI Standards and Data Sets. Translations of any specific International SCI Data Set can be accomplished by translation from the English version into the target language, and be followed by a back-translation into English, to confirm that the original meaning has been preserved. Another approach is to have the initial translation performed by translators who have knowledge of SCI, and afterwards controlled by other person(s) with the same kind of knowledge. The translation process includes both language translation and cultural adaptation, and therefore shall not be made word for word, but will strive to include conceptual equivalence. At a minimum, the inter-rater reliability should be tested by no less than two independent observers, and preferably in multiple countries. Translations must include information on the name, role and background of everyone involved in the translation process, and shall be dated and noted with a version number. By following the proposed guidelines, translated data sets should assure comparability of data acquisition across countries and cultures. If the translation process identifies irregularities or misrepresentation in either the original English version or the target language, the working group for the particular International SCI Data Set shall revise the data set accordingly, which may include re-wording of the original English version in order to accomplish a compromise in the content of the data set.


Crosstalk between the TGF-β and WNT signalling pathways during cardiac fibrogenesis.
Cardiac fibrosis is referred to as an excessive accumulation of stromal cells and extracellular matrix proteins in the myocardium. Progressive fibrosis causes stiffening of the cardiac tissue and affects conduction of electrical impulses, leading to heart failures in a broad range of cardiac conditions. At the cellular level, activation of the cardiac stromal cells and myofibroblast formation are considered as hallmarks of fibrogenesis. At the molecular level, transforming growth factor β (TGF-β) is traditionally considered as a master regulator of the profibrotic processes. More recently, the WNT signalling pathway has also been found to be implicated in the development of myocardial fibrosis. In this review, we summarize current knowledge on the involvement of TGF-β and WNT downstream molecular pathways to cardiac fibrogenesis and describe a crosstalk between these two profibrotic pathways. TGF-β and WNT ligands bind to different receptors and trigger various outputs. However, a growing body of evidence points to cross-regulation between these two pathways. It has been recognized that in cardiac pathologies TGF-β activates WNT/β-catenin signalling, which in turn stabilizes the TGF-β/Smad response. Furthermore both, the non-canonical TGF-β and non-canonical WNT signalling pathways, activate the same mitogen-activated protein kinases (MAPKs): the extracellular signal-regulated kinase (Erk), the c-Jun N-terminal kinases (JNKs) and p38. The crosstalk between TGF-β and WNT pathways seems to play an essential role in switching on the genetic machinery initiating profibrotic changes in the heart. Better understanding of these mechanisms will open new opportunities for development of targeted therapeutic approaches against cardiac fibrosis in the future.


Lotus japonicus LjKUP is induced late during nodule development and encodes a potassium transporter of the plasma membrane.
The KUP family of potassium transporters in plants is large but poorly characterized. We isolated and characterized the first KUP transporter from a legume, LjKUP of Lotus japonicus. Although expressed throughout plants, LjKUP transcript levels were highest in nodules. Induction of LjKUP expression occurred late during nodule development, at a time of rapid organ expansion. A high level of LjKUP expression was maintained in mature, full-sized nodules. However, induction of LjKUP expression was independent of symbiotic nitrogen fixation (SNF), and occurred in ineffective nodules resulting from mutations in either the plant or its microsymbiont, Mesorhizobium loti. Heterologous expression of LjKUP in Escherichia coli showed that the protein is able to transport potassium. Transient expression of a GFP-LjKUP fusion protein in Arabidopsis cells indicated a plasma membrane location for the transporter. Taken together, the results indicate that LjKUP is a potassium transporter of the plasma membrane, which may play roles in cell expansion during nodule development and in ion homeostasis during SNF.


Molecular characterizations of derivatives of HCT116 colorectal cancer cells that are resistant to the chemotherapeutic agent 5-fluorouracil.
5-Fluorouracil (5-FU) is the chemotherapeutic drug of choice for the treatment of metastatic colorectal cancer, but resistance to 5-FU remains a major obstacle to successful therapy. We generated 5-FU-resistant derivatives of the HCT116 human colon cancer cell line by serial passage of these cells in the presence of increasing 5-FU concentrations in an attempt to elucidate the biological mechanisms involved in resistance to 5-FU. Two resultant resistant derivatives, HCT116 ResB and ResD, were characterized for resistance phenotypes, genotypes, and gene expression using cells maintained long-term in 5-FU-free media. Compared to parental HCT116 cells that respond to 5-FU challenge by inducing high levels of apoptosis, ResB and ResD derivatives had significantly reduced apoptotic fractions when transiently challenged with 5-FU. ResB and ResD cells were respectively 27- and 121-fold more resistant to 5-FU, had increased doubling times, and significantly increased plating efficiencies compared to the parental cells. Both resistant derivatives retained the wild-type TP53 genotype, TP53 copy number and CGH profile characteristic of the parental line. Alterations in gene expression in the resistant derivatives compared to the parental line were assessed using oligonucleotide microarrays. Overall, the 5-FU-resistant derivatives were characterized by reduced apoptosis and a more aggressive growth phenotype, consistent with the observed up-regulation of apoptosis-inhibitory genes (e.g., IRAK1, MALT1, BIRC5), positive growth-regulatory genes (e.g., CCND3, CCNE2, CCNF, CYR61), and metastasis genes (e.g., LMNB1, F3, TMSNB), and down-regulation of apoptosis-promoting genes (e.g., BNIP3, BNIP3L, FOXO3A) and negative growth-regulatory genes (e.g., AREG, CCNG2, CDKN1A, CDKN1C, GADD45A). 5-FU metabolism-associated genes (e.g., TYMS, DTYMK, UP) and DNA repair genes (e.g., FEN1, FANCG, RAD23B) were also up-regulated in one or both resistant derivatives, suggesting that the resistant derivatives might be able to overcome both 5-FU inhibition of thymidylate synthase and the DNA damage caused by 5-FU, respectively. Development of 5-FU resistance thus appears to encompass deregulation of apoptosis-, proliferation-, DNA repair-, and metastasis-associated regulatory pathways.


The Bohr Effect Is Not a Likely Promoter of Renal Preglomerular Oxygen Shunting.
The aim of this study was to evaluate whether possible preglomerular arterial-to-venous oxygen shunting is affected by the interaction between renal preglomerular carbon dioxide and oxygen transport. We hypothesized that a reverse (venous-to-arterial) shunting of carbon dioxide will increase partial pressure of carbon dioxide and decrease pH in the arteries and thereby lead to increased oxygen offloading and consequent oxygen shunting. To test this hypothesis, we employed a segment-wise three-dimensional computational model of coupled renal oxygen and carbon dioxide transport, wherein coupling is achieved by shifting the oxygen-hemoglobin dissociation curve in dependence of local changes in partial pressure of carbon dioxide and pH. The model suggests that primarily due to the high buffering capacity of blood, there is only marginally increased acidity in the preglomerular vasculature compared to systemic arterial blood caused by carbon dioxide shunting. Furthermore, effects of carbon dioxide transport do not promote but rather impair preglomerular oxygen shunting, as the increase in acidity is higher in the veins compared to that in the arteries. We conclude that while substantial arterial-to-venous oxygen shunting might take place in the postglomerular vasculature, the net amount of oxygen shunted at the preglomerular vasculature appears to be marginal.


Interdependence of mycobacterial iron regulation, oxidative-stress response and isoniazid resistance.
Iron is an essential cofactor for vital functions in microorganisms. Bacterial pathogens have developed efficient iron-acquisition systems to counteract the defensive sequestration of iron by their hosts. In mycobacteria, the recently described protein, IdeR, negatively controls iron-uptake systems. This protein also has a role in the oxidative-stress response, as well as in resistance to the frontline antimycobacterial drug isoniazid.


Modelling Animal Interactive Rhythms in Communication.
Time is one crucial dimension conveying information in animal communication. Evolution has shaped animals' nervous systems to produce signals with temporal properties fitting their socio-ecological niches. Many quantitative models of mechanisms underlying rhythmic behaviour exist, spanning insects, crustaceans, birds, amphibians, and mammals. However, these computational and mathematical models are often presented in isolation. Here, we provide an overview of the main mathematical models employed in the study of animal rhythmic communication among conspecifics. After presenting basic definitions and mathematical formalisms, we discuss each individual model. These computational models are then compared using simulated data to uncover similarities and key differences in the underlying mechanisms found across species. Our review of the empirical literature is admittedly limited. We stress the need of using comparative computer simulations - both before and after animal experiments - to better understand animal timing in interaction. We hope this article will serve as a potential first step towards a common computational framework to describe temporal interactions in animals, including humans.


Evolutionary history of rabies in Ghana.
Rabies virus (RABV) is enzootic throughout Africa, with the domestic dog (Canis familiaris) being the principal vector. Dog rabies is estimated to cause 24,000 human deaths per year in Africa, however, this estimate is still considered to be conservative. Two sub-Saharan African RABV lineages have been detected in West Africa. Lineage 2 is present throughout West Africa, whereas Africa 1a dominates in northern and eastern Africa, but has been detected in Nigeria and Gabon, and Africa 1b was previously absent from West Africa. We confirmed the presence of RABV in a cohort of 76 brain samples obtained from rabid animals in Ghana collected over an eighteen-month period (2007-2009). Phylogenetic analysis of the sequences obtained confirmed all viruses to be RABV, belonging to lineages previously detected in sub-Saharan Africa. However, unlike earlier reported studies that suggested a single lineage (Africa 2) circulates in West Africa, we identified viruses belonging to the Africa 2 lineage and both Africa 1 (a and b) sub-lineages. Phylogeographic Bayesian Markov chain Monte Carlo analysis of a 405 bp fragment of the RABV nucleoprotein gene from the 76 new sequences derived from Ghanaian animals suggest that within the Africa 2 lineage three clades co-circulate with their origins in other West African countries. Africa 1a is probably a western extension of a clade circulating in central Africa and the Africa 1b virus a probable recent introduction from eastern Africa. We also developed and tested a novel reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of RABV in African laboratories. This RT-LAMP was shown to detect both Africa 1 and 2 viruses, including its adaptation to a lateral flow device format for product visualization. These data suggest that RABV epidemiology is more complex than previously thought in West Africa and that there have been repeated introductions of RABV into Ghana. This analysis highlights the potential problems of individual developing nations implementing rabies control programmes in the absence of a regional programme.


Protein kinases curb cell death.
Networks of aspartic acid-directed caspases play a major role in the execution of programmed cell death. Studies have provided evidence that caspases or their substrates are subjected to phosphorylation, which suggests a potential convergence of protein kinase and caspase signaling pathways. Various caspase substrates, but also several procaspases, are protected from cleavage when they are phosphorylated at sites adjacent to caspase cleavage sites. Whereas many distinct protein kinases could potentially protect substrates from caspase-mediated cleavage, a study has identified protein kinase CK2 as the most prominent kinase that exerts a global inhibition of caspase signaling pathways. CK2 is a component of protein kinase networks that are involved in tumors derived from various tissues. Its dysregulation in many cancers, together with its dual function in promoting cell growth and in suppressing apoptosis, is particularly relevant to its oncogenic potential. Thus, this study suggests that the ability of CK2 to contribute to tumorigenesis resides, at least in part, in its ability to phosphorylate caspases or their targets.


DNA damage stabilizes interaction of CSB with the transcription elongation machinery.
The Cockayne syndrome B (CSB) protein is essential for transcription-coupled DNA repair (TCR), which is dependent on RNA polymerase II elongation. TCR is required to quickly remove the cytotoxic transcription-blocking DNA lesions. Functional GFP-tagged CSB, expressed at physiological levels, was homogeneously dispersed throughout the nucleoplasm in addition to bright nuclear foci and nucleolar accumulation. Photobleaching studies showed that GFP-CSB, as part of a high molecular weight complex, transiently interacts with the transcription machinery. Upon (DNA damage-induced) transcription arrest CSB binding these interactions are prolonged, most likely reflecting actual engagement of CSB in TCR. These findings are consistent with a model in which CSB monitors progression of transcription by regularly probing elongation complexes and becomes more tightly associated to these complexes when TCR is active.


Adjuvant treatment of non-small-cell lung cancer: how do we improve the cure rates further?
Surgery remains the initial treatment for patients with early-stage non-small-cell lung cancer (NSCLC). Additional therapy is necessary because of high rates of distant and local disease recurrence after surgical resection. Early trials of adjuvant chemotherapy and postoperative radiation were often plagued by small patient sample size, inadequate surgical staging, and ineffective or antiquated treatment. A 1995 meta-analysis found a nonsignificant reduction in risk of death for postoperative cisplatin-based chemotherapy. Since then, a new generation of randomized phase III trials have been conducted, some of which have reported a benefit for chemotherapy in the adjuvant setting. The role of postoperative radiation therapy remains to be defined. It may not be beneficial in early-stage NSCLC but still may have utility in stage IIIA disease. Improvement in survival outcomes from adjuvant treatment are likely to result from the evaluation of novel agents, identification of tumor markers predictive of disease relapse, and definition of factors that determine sensitivity to therapeutic agents. Some of the molecularly targeted agents such as the angiogenesis and epidermal growth factor receptor inhibitors are being incorporated into clinical trials. Preliminary results with gene-expression profiles and lung cancer proteomics have been promising. These techniques may be used to create prediction models to identify patients at risk for disease relapse. Molecular markers such as ERCC1 may determine response to treatment. All of these innovations will hopefully increase cure rates for lung cancer patients by maximizing the efficacy of adjuvant therapy.


DNA-templated carbon nanotube field-effect transistor.
The combination of their electronic properties and dimensions makes carbon nanotubes ideal building blocks for molecular electronics. However, the advancement of carbon nanotube-based electronics requires assembly strategies that allow their precise localization and interconnection. Using a scheme based on recognition between molecular building blocks, we report the realization of a self-assembled carbon nanotube field-effect transistor operating at room temperature. A DNA scaffold molecule provides the address for precise localization of a semiconducting single-wall carbon nanotube as well as the template for the extended metallic wires contacting it.


Acceptable medication non-adherence: A crowdsourcing study among French physicians for commonly prescribed medications.
Achieving good medication adherence is a major challenge for patients with chronic conditions. Our study aimed to assess the Threshold for Unacceptable Risk of Non-adherence (TURN), defined as the threshold at which physicians consider the health risks incurred by patients due to medication non-adherence unacceptable, for the most commonly prescribed drugs in France. We conducted an online study using a crowdsourcing approach among French general practitioners and specialists from September 2016 to August 2017. Physicians assessed the TURN for various levels of missed doses by evaluating a series of randomly presented clinical vignettes, each presenting a given medication with a given therapeutic indication. For each "drug-indication group" (i.e., all drugs from the same pharmacological class with a similar therapeutic indication): 1) we described the distribution of physicians' assessments, 2) we provided a summary estimate of the TURN, defined as the frequency of missed doses above which 75% of the physicians' assessments were located; 3) we computed the number of pill boxes reimbursed in France in 2016 to put our results into context. We collected a total of 5365 assessments from 544 physicians, each of whom evaluated a random sample among 528 distinct clinical vignettes. Estimates of the TURN varied widely across drug-indication groups, ranging from risk considered unacceptable with 1 daily dose missed per month (e.g., insulin for diabetes) to risk always considered acceptable (e.g., anti-dementia drugs). Drugs with an estimated TURN of over one missing daily dose per week represented 44.9% of the prescription volume of the medications assessed in our study. According to physicians, the impact of non-adherence may vary greatly. Patient-physician discussions on the variable consequences of non-adherence could lead to a paradigm shift by seeking to reach "optimal adherence" depending on drugs rather than unrealistic "perfect adherence" to all drugs.


Complement component C5a promotes expression of IL-22 and IL-17 from human T cells and its implication in age-related macular degeneration.
Age related macular degeneration (AMD) is the leading cause of irreversible blindness in elderly populations worldwide. Inflammation, among many factors, has been suggested to play an important role in AMD pathogenesis. Recent studies have demonstrated a strong genetic association between AMD and complement factor H (CFH), the down-regulatory factor of complement activation. Elevated levels of complement activating molecules including complement component 5a (C5a) have been found in the serum of AMD patients. Our aim is to study whether C5a can impact human T cells and its implication in AMD. Human peripheral blood mononuclear cells (PBMCs) were isolated from the blood of exudative form of AMD patients using a Ficoll gradient centrifugation protocol. Intracellular staining and enzyme-linked immunosorbent assays were used to measure protein expression. Apoptotic cells were detected by staining of cells with the annexin-V and TUNEL technology and analyzed by a FACS Caliber flow cytometer. SNP genotyping was analyzed by TaqMan genotyping assay using the Real-time PCR system 7500. We show that C5a promotes interleukin (IL)-22 and IL-17 expression by human CD4+ T cells. This effect is dependent on B7, IL-1β and IL-6 expression from monocytes. We have also found that C5a could protect human CD4+ cells from undergoing apoptosis. Importantly, consistent with a role of C5a in promoting IL-22 and IL-17 expression, significant elevation in IL-22 and IL-17 levels was found in AMD patients as compared to non-AMD controls. Our results support the notion that C5a may be one of the factors contributing to the elevated serum IL-22 and IL-17 levels in AMD patients. The possible involvement of IL-22 and IL-17 in the inflammation that contributes to AMD may herald a new approach to treat AMD.