Rapid acid treatment of Escherichia coli: transcriptomic response and recovery.
Many E. coli genes show pH-dependent expression during logarithmic growth in acid (pH 5-6) or in base (pH 8-9). The effect of rapid pH change, however, has rarely been tested. Rapid acid treatment could distinguish between genes responding to external pH, and genes responding to cytoplasmic acidification, which occurs transiently following rapid external acidification. It could reveal previously unknown acid-stress genes whose effects are transient, as well as show which acid-stress genes have a delayed response. Microarray hybridization was employed to observe the global gene expression of E. coli K-12 W3110 following rapid acidification of the external medium, from pH 7.6 to pH 5.5. Fluorimetric observation of pH-dependent tetR-YFP showed that rapid external acidification led to a half-unit drop in cytoplasmic pH (from pH 7.6 to pH 6.4) which began to recover within 20 s. Following acid treatment, 630 genes were up-regulated and 586 genes were down-regulated. Up-regulated genes included amino-acid decarboxylases (cadA, adiY, gadA), succinate dehydrogenase (sdhABCD), biofilm-associated genes (bdm, gatAB, and ymgABC), and the Gad, Fur and Rcs regulons. Genes with response patterns consistent with cytoplasmic acid stress were revealed by addition of benzoate, a membrane-permeant acid that permanently depresses cytoplasmic pH without affecting external pH. Several genes (yagU, ygiN, yjeI, and yneI) were up-regulated specifically by external acidification, while other genes (fimB, ygaC, yhcN, yhjX, ymgABC, yodA) presented a benzoate response consistent with cytoplasmic pH stress. Other genes (the nuo operon for NADH dehydrogenase I, and the HslUV protease) showed delayed up-regulation by acid, with expression rising by 10 min following the acid shift. Transcriptomic profiling of E. coli K-12 distinguished three different classes of change in gene expression following rapid acid treatment: up-regulation with or without recovery, and delayed response to acid. For eight genes showing acid response and recovery (fimB, ygaC, yhcN, yhjX, ymgABC, yodA), responses to the permeant acid benzoate revealed expression patterns consistent with sensing of cytoplasmic pH. The delayed acid response of nuo genes shows that NADH dehydrogenase I is probably induced as a secondary result of acid-associated metabolism, not as a direct response to cytoplasmic acidification.


Thermodynamic limits on drug loading in nanoparticle cores.
Recently, biodegradable nanoparticles based on block copolymers have attracted attention as effective drug delivery vehicles. Maximizing the amount of drug loaded into particle is the desired goal, but loadings of only between 3 to about 25 wt% drug (for paclitaxel) are found experimentally. The reasons for the low loading and variability in loading have not been fully explained. In this study, a model is presented that quantitatively explains the observed phenomena. The thermodynamic model of drug loading is based on the molar free energy of the drug, which depends on the block copolymers size (entropic term), the interaction parameter between the drug and the hydrophobic core (enthalpic term), and the pressure-volume work to load the particle. The pressure-volume work, related directly to the interfacial tension between the core and the corona region, has not been previously considered with respect to drug loading. To validate the model, calculations were compared with experimental results for organic solutes, including paclitaxel, loaded into poly(ethylene glycol)-b-poly(epsilon-caprolactone), PEG-b-PCL block copolymer micelles. The model developed was found to predict the loading values in close agreement with experiments reported in the literature.


C26-Ceramide as highly sensitive biomarker for the diagnosis of Farber Disease.
Farber disease (FD) is a rare autosomal recessive disease caused by mutations in the acid ceramidase gene (ASAH1). Low ceramidase activity results in the accumulation of fatty substances, mainly ceramides. Hallmark symptoms at clinical level are periarticular nodules, lipogranulomas, swollen and painful joints and a hoarse voice. FD phenotypes are heterogeneous varying from mild to very severe cases, with the patients not surviving past their first year of life. The diagnostic aspects of FD are poorly developed due to the rarity of the disease. In the present study, the screening for ceramides and related molecules was performed in Farber affected patients (n = 10), carriers (n = 11) and control individuals (n = 192). This study has the highest number of enrolled Farber patients and carriers reported to present. Liquid chromatography multiple reaction mass spectrometry (LC/MRM-MS) studies revealed that the ceramide C26:0 and especially its isoform 1 is a highly sensitive and specific biomarker for FD (p < 0.0001). The new biomarker can be determined directly in the dried blood spot extracts with low sample consumption. This allows for easy sample preparation, high reproducibility and use in high throughput screenings.


Social readjustment and ischemic stroke: lack of an association in a multiethnic population.
Clinical experience has suggested that stressful life events and ongoing stressful illness, collectively termed 'social readjustment', may precipitate stroke. We investigated the association between a simple in-office evaluation of such stressors and stroke in an urban, multiethnic study population. Cases were patients from the Northern Manhattan Stroke Study with first ischemic stroke; controls were derived through random digit dialing with n:m matching for age, gender, and race-ethnicity. Social readjustment was measured through in-person interview using Amster and Krauss' Geriatric Social Readjustment Rating Scale (GSRRS), a one-time, 35-item, checklist type weighted questionnaire of stressful life events occurring in the previous 6 months. Conditional logistic regression was used to analyze the GSRRS and its quartiles as well as stressful events subgroups, adjusting for education, hypertension, cardiac disease, diabetes, and number of weekly visits as a measure of socialization. Six hundred and fifty-five cases of ischemic stroke and 1,087 controls were utilized. The mean age of the cases was 69.8 years, with 55.4% women, 51.0% Hispanics, 28.4% blacks, and 19.1% whites. GSRRS scores ranged from 0 to 812; the mean score was 205.5 for the cases and 206.2 for the controls. The analysis showed no association between stroke and a 20-point increase on the GSRRS (OR = 1.01, 95% CI = 0.99-1.01). There was also no effect for the second, third or highest versus lowest quartile. No association was found in age, gender or race-ethnic subgroups, or when analyzing negative events, severely threatening events, or ongoing stressful illnesses separately. While this study does not preclude social readjustment as a stroke risk factor, it suggests that the one-time assessment often done in the medical office setting has little relevance for stroke prevention planning.


Kinetics of the reaction of the simplest Criegee intermediate with ammonia: a combination of experiment and theory.
The kinetics of the reaction of the simplest Criegee intermediate (CH2OO) with ammonia has been measured under pseudo-first-order conditions with two different experimental methods. We investigated the rate coefficients at 283, 298, 308, and 318 K at a pressure of 50 Torr using an OH laser-induced fluorescence (LIF) method. Weak temperature dependence of the rate coefficient was observed, which is consistent with the theoretical activation energy of -0.53 kcal mol-1 predicted by quantum chemistry calculation at the QCISD(T)/CBS//B3LYP/6-311+G(2d,2p) level. At 298 K, the rate coefficient at 50 Torr from the OH LIF experiment was (5.64 ± 0.56) × 10-14 cm3 molecule-1 s-1 while at 100 Torr we obtained a slightly larger value of (8.1 ± 1.0) × 10-14 cm3 molecule-1 s-1 using the UV transient absorption method. These experimental values are within the theoretical error bars of the present as well as previous theoretical results. Our experimental results confirmed the previous conclusion that ammonia is negligible in the consumption of CH2OO in the atmosphere. We also note that CH2OO may compete with OH in the oxidation of ammonia under certain circumstances, such as at night-time, high altitude and winter time.


Long-term (6-wk) hindlimb suspension inhibits spermatogenesis in adult male rats.
The International Space Station will allow extended habitation in space and long-term exposure to microgravity (microG). A concern is the impact of long-term microG exposure on the ability of species to reproduce. The model often used to simulate microG is rat hindlimb suspension (HLS), where the hindlimbs are elevated above the cage floor with a tail harness. Experiments described here are the first to examine the effect of long-term HLS on testicular function in adult male rats. Free-roaming (controls), animals with only the tail harnessed but hindlimbs in contact with the cage floor (TO), and HLS animals were tested for 6 wk. Cryptorchidism was prevented in TO and HLS animals by partial constriction of the inguinal canal with sutures. All parameters were compared at the end of the 6-wk experiment. Testicular weights and spermatogenesis were significantly reduced by HLS, such that no spermatogenic cells beyond round spermatids were present and epididymides were devoid of mature sperm. In many tubules, loss of all germ cells, except a few spermatogonia, resulting in histopathology similar to the Sertoli cell, was observed. Spermatogenesis appeared unaffected in control and TO animals. Sertoli and Leydig cell appearance, testosterone, luteinizing hormone, and follicle-stimulating hormone levels, and epididymal and seminal vesicle weight were unchanged by HLS. Cortisone was not elevated by HLS; thus stress may not be a factor. These results demonstrate that spermatogenesis is severely inhibited by long-term HLS, whereas testicular androgen production is not. These results have significant implications regarding serious effects of long-term exposure to microG on the reproductive capability of scrotal mammals, including humans.


Intracellular replication of attenuated Mycobacterium tuberculosis phoP mutant in the absence of host cell cytotoxicity.
Intracellular pathogen Mycobacterium tuberculosis survives and replicates in macrophages but limited information is available on its replication into non-phagocytic cells. Here we study the role of the M. tuberculosis virulence gene phoP in the intracellular growth with rat and human lung fibroblasts. In contrast to macrophages, attenuated M. tuberculosis phoP mutant was able to multiply intracellularly in fibroblasts at the same level as the virulent M. tuberculosis. However, when M. tuberculosis virulence was studied using human foetal lung fibroblasts, MRC-5 cell line, the virulent strain caused a significant damage in cells compared with attenuated strains BCG and M. tuberculosis phoP mutant. We analysed the effect of cytoskeleton inhibitors in NRK-49F fibroblasts. M. tuberculosis invasion was not inhibited, suggesting that mycobacterial uptake was microtubule and microfilament independent. Our results suggest that PhoP in M. tuberculosis does not regulate intracellular replication in fibroblasts, contrary to what happens in macrophages. The ability of M. tuberculosis phoP mutant to replicate within non-phagocytic cells, such as fibroblasts, without causing damage, could be a potential advantage for a live attenuated vaccine against tuberculosis.


Genetic diversity of human immunodeficiency virus type 2: evidence for distinct sequence subtypes with differences in virus biology.
The virulence properties of human immunodeficiency virus type 2 (HIV-2) are known to vary significantly and to range from relative attenuation in certain individuals to high-level pathogenicity in others. These differences in clinical manifestations may, at least in part, be determined by genetic differences among infecting virus strains. Evaluation of the full spectrum of HIV-2 genetic diversity is thus a necessary first step towards understanding its molecular epidemiology, natural history of infection, and biological diversity. In this study, we have used nested PCR techniques to amplify viral sequences from the DNA of uncultured peripheral blood mononuclear cells from 12 patients with HIV-2 seroreactivity. Sequence analysis of four nonoverlapping genomic regions allowed a comprehensive analysis of HIV-2 phylogeny. The results revealed (i) the existence of five distinct and roughly equidistant evolutionary lineages of HIV-2 which, by analogy with HIV-1, have been termed sequence subtypes A to E; (ii) evidence for a mosaic HIV-2 genome, indicating that coinfection with genetically divergent strains and recombination can occur in HIV-2-infected individuals; and (iii) evidence supporting the conclusion that some of the HIV-2 subtypes may have arisen from independent introductions of genetically diverse sooty mangabey viruses into the human population. Importantly, only a subset of HIV-2 strains replicated in culture: all subtype A viruses grew to high titers, but attempts to isolate representatives of subtypes C, D, and E, as well as the majority of subtype B viruses, remained unsuccessful. Infection with all five viral subtypes was detectable by commercially available serological (Western immunoblot) assays, despite intersubtype sequence differences of up to 25% in the gag, pol, and env regions. These results indicate that the genetic and biological diversity of HIV-2 is far greater than previously appreciated and suggest that there may be subtype-specific differences in virus biology. Systematic natural history studies are needed to determine whether this heterogeneity has clinical relevance and whether the various HIV-2 subtypes differ in their in vivo pathogenicity.


Mechanism of action of 1,25-dihydroxyvitamin D3 on intestinal calcium absorption.
1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is the major controlling hormone of intestinal calcium absorption. As the body's demand for calcium increases from a diet deficient in calcium, from growth, pregnancy or lactation, the synthesis of 1,25(OH)(2)D(3) is increased resulting in the stimulation of intestinal calcium absorption. However a complete description of the molecular mechanisms involved in the 1,25(OH)(2)D(3) regulated calcium absorptive process remains incomplete. Intestinal calcium absorption occurs by both an active saturable transcellular pathway and a passive nonsaturable paracellular pathway. Each step in the process of transcellular calcium transport (apical entry of calcium, translocation of calcium through the interior of the enterocyte and basolateral extrusion of calcium by the plasma membrane pump) has been reported to involve a vitamin D dependent component. This article will review recent studies, including those using knockout mice, that have suggested that 1,25(OH)(2)D(3) mediated calcium absorption is more complex than the traditional three step model of transcellular calcium transport. Current concepts are reviewed and questions that remain are addressed. Evidence for a role of 1,25(OH)(2)D(3) in the regulation of the paracellular pathway is also discussed.


Ketoconazole potentiates terfenadine-induced apoptosis in human Hep G2 cells through inhibition of cytochrome p450 3A4 activity.
Terfenadine (TF) is a highly potent histamine H1 receptor antagonist that in clinically effective doses is free of significant central nervous system side effects. Ketoconazole (KT) is a worldwide used oral antifungal agent with a broad spectrum of activity against both superficial and systemic mycosis. Simultaneously administration of KT and TF has been reported to induce several potent symptoms including cardiotoxicity, excitotoxicity, inhibition of blood mononuclear cells proliferation, and cardiovascular toxicity. However, the intracellular molecular mechanisms of TF-KT interactions in cells were still uncertain. In this study, we first demonstrated that TF (5-30 microM) induced apoptosis in several types of human cancer cell lines including human hepatoma (Hep G2), colorectal cancer (COLO 205), and fibroblast (CCD 922SK) cells for 24 h. The cellular responses to TF-induced apoptosis were demonstrated to be associated with the p53-signaling pathway, including induction of p53, p21/Cip1, p27/Kip1, bax protein expression and inhibition of bcl-2 protein expression. To realized the role of H1 receptor involved in TF-induced apoptosis, different H1 receptor antagonists including promethazine, mequitazine, and chlorpheniramin (50-100 microM) were administered and demonstrated that these chemicals cannot induced apoptosis through the H1 receptor signaling pathway. Interestingly, we found that the apoptotic effect of TF (2.5 microM) was significantly potentiated by KT (1 microM) treatment in Hep G2 cells through inhibition of the cytochrome p450 3A4 (CYP 3A4) activity. Such results were demonstrated by decreased of the TF activity with recombinant CYP 3A4, which prepared from baculovirus-infected insect cells. Our results provide the molecular basis of TF-KT interaction and this information should allow more rational forecasting of the risk for TF therapy during co-administration of KT.


The prophylactic effects of different Lactobacilli on collagen-induced arthritis in rats.
Recent studies have shed light on the prophylactic effects of Lactobacilli on rheumatoid arthritis (RA). However, the modulatory mechanisms of Lactobacilli remain unclear. The current study evaluated different Lactobacillus species' ability to alleviate arthritis induced by collagen. Rats were intragastrically administered different lactobacilli cocktails two weeks before arthritis induction. The results revealed that the performance of Lactobacillus in relieving arthritis was different for some species. L. reuteri, L. casei, L. rhamnosus and L. fermentum attenuated RA through species-independent pathways that inhibited pro-inflammatory cytokines and anti-CII-antibodies; and through species-dependent immune regulation that was based on rebalancing the intestinal microbiota, and metabolites such as short-chain fatty acids. In particular, L. reuteri and L. casei weaken the Th1 immune response, while L. rhamnosus and L. fermentum impaired Th17 responses. Interestingly, L. plantarum did not alleviate arthritis although it did suppress Th1 and Th17 immune responses, while L. salivarius only delayed the onset of arthritis without influencing the immune response. In conclusion, Lactobacilli protect against collagen-induced-arthritis through both common and individual pathways.


Enhanced immune response after a second dose of an AS03-adjuvanted H1N1 influenza A vaccine in patients after hematopoietic stem cell transplantation.
Seroconversion rates following influenza vaccination in patients with hematologic malignancies after hematopoietic stem cell transplantation (HSCT) are known to be lower compared to healthy adults. The aim of our diagnostic study was to determine the rate of seroconversion after 1 or 2 doses of a novel split virion, inactivated, AS03-adjuvanted pandemic H1N1 influenza vaccine (A/California/7/2009) in HSCT recipients (ClinicalTrials.gov Identifier: NCT01017172). Blood samples were taken before and 21 days after a first dose and 21 days after a second dose of the vaccine. Antibody (AB) titers were determined by hemagglutination inhibition assay. Seroconversion was defined by either an AB titer of ≤ 1:10 before and ≥ 1:40 after or ≥ 1:10 before and ≥ 4-fold increase in AB titer 21 days after vaccination. Seventeen patients (14 allogeneic, 3 autologous HSCT) received 1 dose and 11 of these patients 2 doses of the vaccine. The rate of seroconversion was 41.2% (95% confidence interval [CI] 18.4-67.1) after the first and 81.8% (95% CI 48.2-97.7) after the second dose. Patients who failed to seroconvert after 1 dose of the vaccine were more likely to receive any immunosuppressive agent (P = .003), but time elapsed after or type of HSCT, age, sex, or chronic graft-versus-host disease was not different when compared to patients with seroconversion. In patients with hematologic malignancies after HSCT the rate of seroconversion after a first dose of an adjuvanted H1N1 influenza A vaccine was poor, but increased after a second dose.


Viral oncoproteins E1A and E7 and cellular LxCxE proteins repress SUMO modification of the retinoblastoma tumor suppressor.
The retinoblastoma tumor suppressor protein (pRB) is a major regulator of cell-cycle progression and cellular differentiation. Central to pRB function is the pocket domain, which serves as the main binding region for cellular regulators. In tumors pRB is frequently inactivated by mutations in the pocket domain or by binding of viral oncoproteins to this region. A characteristic feature of these viral oncoproteins and many cellular pRB-binding partners is an LxCxE sequence motif, which interacts with pRB's pocket domain. Here, we show that the ubiquitin-like modifier SUMO is covalently attached to a distinct residue (K720) of pRB within the B-box of the pocket region that binds LxCxE-motif proteins. We provide evidence that SUMO preferentially targets the active, hypophosphorylated form of pRB and show that tumorigenic mutations of pRB in the pocket domain lead to a loss of SUMOylation. Notably, the level of pRB SUMOylation is controlled by the interaction of pRB with viral and cellular LxCxE-motif proteins. Inhibitors of pRB function, including the viral oncoproteins E1A and E7 and the cellular E1A-like inhibitor of differentiation EID-1, completely abolish SUMO modification of pRB. Conversely, pRB mutants deficient in binding of LxCxE-motif proteins exhibit a drastically enhanced modification by SUMO. Finally, we provide evidence that SUMOylation can influence pRB function, as the SUMO-deficient pRB(K720R) mutant exerts a slightly higher repressive potential on an E2F-responsive reporter gene than wild-type pRB. Taken together, these data identify SUMO modification as a novel post-translational modification of pRB that may control pRB activity by modulating LxCxE-pocket interactions.


Hypertrophic lichen sclerosus with dyskeratosis and parakeratosis--a common presentation of vulvar lichen sclerosus not associated with a significant risk of malignancy.
Epithelial hyperplasia, individual necrotic keratocytes, and parakeratosis are common findings in lichen sclerosus. When those changes are prominent, they may pose diagnostic problems, especially because such lesions often show no or only minimal sclerosis. Necrotic keratocytes are often numerous and are found in all reaches of the epidermis, presenting themselves as eosinophilic globules with or without remnants of pyknotic nuclei. Because those changes tend to be accentuated focally above dermal papillae, they often give rise to narrow columns of parakeratosis in the overlying cornified layer. Within those columns, individual necrotic keratocytes with pyknotic nuclei are preserved as distinct dyskeratotic parakeratotic cells. That constellation of findings is fairly characteristic of hypertrophic lichen sclerosus. It was found, at least subtle and focally, in 14 of 70 consecutive biopsy specimens of lichen sclerosus, most of which came from the vulva of elderly women. Although similar cases have been described as differentiated vulvar intraepithelial neoplasia (VIN) in the literature, there was no significant nuclear atypia, no crowding of nuclei, and no significant mitotic activity in any of those lesions. Follow-up of at least 5 years in 8 patients revealed no development of squamous cell carcinoma. Hypertrophic lichen sclerosus with dyskeratosis and parakeratosis seems to be a relatively common presentation of vulvar lichen sclerosus not associated with a significant risk of malignancy.


Correlation of radiographic muscle cross-sectional area with glenohumeral deformity in children with brachial plexus birth palsy.
Muscle imbalance about the shoulder in children with persistent brachial plexus birth palsy is thought to contribute to glenohumeral joint deformity. We quantified cross-sectional areas of the internal and external rotator muscles in the shoulder by magnetic resonance imaging in patients with chronic brachial plexopathy and the correlation between these muscle cross-sectional area ratios and glenohumeral deformity. The purposes of this investigation were to evaluate differences in the ratios between affected and unaffected shoulders in the same individual and to assess whether an increased internal to external rotator muscle cross-sectional area correlated with greater glenohumeral deformity. This cohort study consisted of magnetic resonance imaging of seventy-four patients with chronic neuropathic changes about the shoulder from brachial plexus birth palsy. There were at least nine patients with scans available for each of the five classified subtypes of glenohumeral deformity: type I (fifteen patients), type II (seventeen), type III (seventeen), type IV (sixteen), and type V (nine). Cross-sectional areas of the pectoralis major, teres minor-infraspinatus (external rotators), and subscapularis muscles were measured. The supraspinatus muscle cross-sectional area could not be reliably measured. The ratio of subscapularis to external rotators, the ratio of pectoralis major to external rotators, and the compound ratio of subscapularis and pectoralis major to external rotators were compared with the severity of the glenohumeral deformity. Passive range of motion, Mallet and Toronto clinical scores, and Narakas type were also compared with the severity of the glenohumeral deformity and the muscle cross-sectional area measurements. Muscle cross-sectional area ratios were significantly correlated with glenohumeral deformity type. The mean ratio of pectoralis major to external rotators for affected shoulders over all deformity types compared with that for unaffected shoulders was significantly increased by 30% (p < 0.001); the mean ratio for subscapularis and pectoralis major to external rotators, by 19% (p = 0.015), and the mean ratio for subscapularis to external rotators, by 10% (p = 0.008). There was a significant increase in the ratio of pectoralis major to external rotators in affected shoulders within each type of deformity. Analysis of variance indicated higher ratios of pectoralis major to external rotator muscle cross-sectional areas in more severe deformity types (p < 0.001). There were significant differences in external rotation measurements with the shoulder at 90 degrees of abduction only among glenohumeral deformity types I, II, and III (p < 0.05). The degree of muscle imbalance between internal and external rotators about the shoulder is measurable by magnetic resonance imaging in children with persistent brachial plexopathy, and the imbalance correlates with the degree of glenohumeral deformity. Our results may provide useful information to guide the timing and the choice of operative intervention in these children.


Oral cavity and base of the tongue tumors. Correlation between clinical, MRI and pathological staging of primary tumor.
The diagnosis of oral cavity and oropharyngeal tumors can be obtained through clinical examination and biopsy. CT and MRI can then be used to define the extension of the disease. The aim of this study was to define the accuracy of clinical and MRI T staging of oral cavity and base of the tongue tumors and correlate the results with pathological data. Mandibular involvement, in a subgroup of patients, was determined and sensitivity, specificity, accuracy and positive and negative predictive values were evaluated. Fifty-nine patients affected by squamous cell carcinoma and 1 case of adenoido-cystic carcinoma were examined by means of a superconductive MR unit, using SE T1, and fat-suppressed T2 weighted sequences before contrast medium infusion. SE T1 and T1 fat-suppressed sequences after gadolinium-DTPA infusion were used. T stage accuracy of both clinical examination and MRI were found to be respectively 62% (k 0.459) and 82% (k 0.775). The sensitivity, specificity and accuracy of MRI in the detection of mandibular involvement were 94.1%, 60% and 81.5%, while the positive and negative predictive values were 80% and 85.7%, respectively. The sensitivity, specificity and accuracy of clinical examination in the detection of mandibular involvement were 100%, 30% and 74.1%, while the positive and negative predictive values were 70.8% and 100%. In the present study, MRI was seen to be an adequate technique for the assessment of oral cavity malignancies, in the evaluation of depth invasion, presence and extension of mandibular involvement.


Code stroke. An attempt to shorten inhospital therapeutic delays.
Significant delays often occur during the treatment of patients with acute stroke. Some of those delays occur in the hospital. We attempted to shorten inhospital treatment intervals by creating a rapid-response system, similar to that available for cardiac arrest, that would allow the stroke team to be available within a few minutes to care for stroke victims. We connected all beepers (pocket pagers) of stroke team members to a common access number and instructed the emergency staff to activate that number immediately upon arrival of a stroke victim. We monitored the response time and treatment interval for patients who were treated after this system was activated (Code Stroke patients) during the first 3 months of its availability and compared the results to those of patients seen for similar reasons during the study period but without the use of Code Stroke (control patients). A total of 12 Code Stroke patients were available for analysis, representing 12% of all patients (n = 98) seen in the emergency department for ischemic stroke during the study period. The remaining 86 patients constituted the control group. The mean time to evaluation of a Code Stroke patient by a stroke team member was 4.8 minutes (range, 2 to 7 minutes), and the mean time to treatment institution was 30 minutes (range, 10 to 120 minutes). There were significant differences between the consultation intervals in the two groups (P < .05). There was only a trend of a difference between treatment institution intervals (P = .06). It is possible to shorten inhospital treatment delays by instituting rapid-response systems within individual institutions.


The incidence of dementia and intake of animal products: preliminary findings from the Adventist Health Study.
We investigated the relationship between animal product consumption and evidence of dementia in two cohort substudies. The first enrolled 272 California residents matched for age, sex, and zip code (1 vegan, 1 lacto-ovo-vegetarian, and 2 'heavy' meat eaters in each of 68 quartets). This design ensured a wide range of dietary exposure. The second included 2,984 unmatched subjects who resided within the Loma Linda, California area. All subjects were enrolled in the Adventist Health Study. The matched subjects who ate meat (including poultry and fish) were more than twice as likely to become demented as their vegetarian counterparts (relative risk 2.18, p = 0.065) and the discrepancy was further widened (relative risk 2.99, p = 0.048) when past meat consumption was taken into account. There was no significant difference in the incidence of dementia in the vegetarian versus meat-eating unmatched subjects. There was no obvious explanation for the difference between the two substudies, although the power of the unmatched sub-study to detect an effect of 'heavy' meat consumption was unexpectedly limited. There was a trend towards delayed onset of dementia in vegetarians in both substudies.


Muscle volume changes.
Measurements have been made of the volume changes accompanying single isometric and isotonic twitches of frog sartorius muscle. The volume change consists of a rapid increase, a subsequent decrease, and a return to the initial volume; the order of magnitude of increase and decrease is 10(-5) cc/g of muscle. This volume change is length-dependent: the initial increase becomes more pronounced as the initial length of the muscle is decreased, while the volume decrease is greatest at reference length and is diminished for longer and shorter initial lengths. Muscle volume changes are also dependent upon temperature and amount of shortening: the return phase is prolonged as the temperature is lowered; and, in an isotonic twitch, a volume increase accompanying muscle shortening is superimposed upon the volume change described for an isometric twitch. These "shortening volume changes" may correspond to the volume decrease observed in frog muscle under a passive stretch. If the active state is prolonged by the use of a frog Ringer solution in which iodide ions have been substituted for chloride ions, the time course of the volume decrease is likewise prolonged; this suggests a relationship between the volume decrease and the active state of the muscle.


Effects of methyl substitution on DNA binding enthalpies of enantiopure Ru(phenanthroline)2dipyridophenazine2+ complexes.
Isothermal titration calorimetry (ITC) has been utilized to investigate the effect of methyl substituents on the intercalating dppz ligand of the enantiomers of the parent complex Ru(phen)2dppz2+ (phen = 1,10-phenanthroline; dppz = dipyrido[3,2-a:2',3'-c]phenazine) on DNA binding thermodynamics. The methylated complexes (10-methyl-dppz and 11,12-dimethyl-dppz) have large, concentration-dependent, positive heats of dilution, and a strong endothermic background is also apparent in the ITC-profiles from titration of methylated complexes into poly(dAdT)2, which make direct comparison between complexes difficult. By augmenting a simple cooperative binding model with one equilibrium for complex self-aggregation in solution and one equilibrium for complex aggregation on saturated DNA, it was possible to find an excellent global fit to the experimental data with DNA affinity parameters restricted to be equal for all Δ-enantiomers as well as for all Λ-enantiomers. In general, enthalpic differences, compared to the unsubstituted complex, were small and less than 4 kJ mol-1, except for the heat of intercalation of Δ-10-methyl-dppz (-11,6 kJ mol-1) and Λ-11,12-dimethyl-dppz (+4.3 kJ mol-1).


Association between human papillomavirus type 16 E6 and E7 variants with subsequent persistent infection and recurrence of cervical high-grade squamous intraepithelial lesion after conization.
The study aimed to detect the variants of human papillomavirus (HPV) type 16 E6 and E7 in patients with cervical high-grade squamous intraepithelial lesion (HSIL), and to determine the existence and recurrence of persistent infection after treatment with loop electrosurgical excision procedure (LEEP). Preoperatively collected cervical exfoliated cells from 100 HPV 16 positive HSIL patients enrolled in the study were used to test for E6 and E7 variants. Follow-ups which included TCT, HPV test, and colposcopy were performed every 3 months after the operation, and colposcopic biopsy and endocervical curettage were performed for patients with abnormalities. Patients were followed for 2 years, and recurrence was defined as detecting low-grade squamous intraepithelial lesion (LSIL) or relapse of HSIL in 1 year. In 81% of patients, the E6 variant was the Asian prototype (As.P), 14% of patients had the European variant, 2% had the European prototype (EP), and 3% had the African 1 variant (Af1). The HPV16 could be easily cleared by LEEP in patients with As.P. Persistent infection or recurrence was very rare in this group. The patients with European variants T350G or A442C had a significantly higher incidence of persistent and recurring HPV16 infection. In conclusion, (i) in most cases, As.P caused HSIL. (ii) The European variant E6 T350G/A442C may be associated with higher rates of recurring and persistent HPV16 infection after the LEEP. (iii) The E7 gene mutation may not be a risk factor for recurring HSIL caused by HPV16 or persistent infection. J. Med. Virol. 88:1982-1988, 2016. © 2016 Wiley Periodicals, Inc.


Gene selection via the BAHSIC family of algorithms.
Identifying significant genes among thousands of sequences on a microarray is a central challenge for cancer research in bioinformatics. The ultimate goal is to detect the genes that are involved in disease outbreak and progression. A multitude of methods have been proposed for this task of feature selection, yet the selected gene lists differ greatly between different methods. To accomplish biologically meaningful gene selection from microarray data, we have to understand the theoretical connections and the differences between these methods. In this article, we define a kernel-based framework for feature selection based on the Hilbert-Schmidt independence criterion and backward elimination, called BAHSIC. We show that several well-known feature selectors are instances of BAHSIC, thereby clarifying their relationship. Furthermore, by choosing a different kernel, BAHSIC allows us to easily define novel feature selection algorithms. As a further advantage, feature selection via BAHSIC works directly on multiclass problems. In a broad experimental evaluation, the members of the BAHSIC family reach high levels of accuracy and robustness when compared to other feature selection techniques. Experiments show that features selected with a linear kernel provide the best classification performance in general, but if strong non-linearities are present in the data then non-linear kernels can be more suitable. Accompanying homepage is http://www.dbs.ifi.lmu.de/~borgward/BAHSIC. Supplementary data are available at Bioinformatics online.


Structure-based Methods for Binding Mode and Binding Affinity Prediction for Peptide-MHC Complexes.
Understanding the mechanisms involved in the activation of an immune response is essential to many fields in human health, including vaccine development and personalized cancer immunotherapy. A central step in the activation of the adaptive immune response is the recognition, by T-cell lymphocytes, of peptides displayed by a special type of receptor known as Major Histocompatibility Complex (MHC). Considering the key role of MHC receptors in T-cell activation, the computational prediction of peptide binding to MHC has been an important goal for many immunological applications. Sequence- based methods have become the gold standard for peptide-MHC binding affinity prediction, but structure-based methods are expected to provide more general predictions (i.e., predictions applicable to all types of MHC receptors). In addition, structural modeling of peptide-MHC complexes has the potential to uncover yet unknown drivers of T-cell activation, thus allowing for the development of better and safer therapies. In this review, we discuss the use of computational methods for the structural modeling of peptide-MHC complexes (i.e., binding mode prediction) and for the structure-based prediction of binding affinity.


1,3-Benzothia-zolium tetra-chlorido-aurate(III) tetra-hydro-furan solvate.
In the crystal structure of the title ionic compound (C(7)H(6)NS)[AuCl(4)]·C(4)H(8)O, the [AuCl(4)](-) anion shows a typical square-planar geometry. Numerous weak C-H⋯Cl hydrogen bonds between [AuCl(4)](-) and the 1,3-benzothia-zolium units form layers comprised of 24-membered rings in which hydrogen-bonded tetra-hydro-furan (THF) solvent mol-ecules are accommodated. C-H⋯Cl inter-actions between THF and [AuCl(4)](-) from adjacent layers result in bilayers. These are further stabilized by π-π inter-actions between the thia-zole and benzene rings [centroid-centroid distance = 3.971 (3) Å], resulting in the formation of a three-dimensional supra-molecular assembly.


The effects of different training programs on the trapezius muscle of women with work-related neck and shoulder myalgia.
The aim of this study was to examine the effects of training on the structural characteristics of the trapezius muscle in women with work-related trapezius myalgia. Muscle biopsies were taken before and after 10 weeks of three different training programs (strength, endurance and coordination). Enzyme-immunohistochemical analysis was performed to assess muscle fibre types, fibre area, capillary supply and cytochrome c oxidase (COX) activity. There was an increase in the proportion of type IIA fibres in strength trained group (P < 0.05). Strength training elicited a preferential increase in the area of type II fibres (P < 0.05); both strength and endurance programs induced an increase in the number of capillaries around type I and IIA muscle fibres. Finally, all training programs induced a decrease in the proportion of COX-negative fibres. In conclusion, the trapezius muscle of women with neck and shoulder myalgia is characterised by a great potential of adaptation to physical exercise over a period of 10 weeks. The significant changes in the number of capillaries and the specific changes induced by training at the level of muscle fibres might well explain the improvement of muscle function.


Effect of reducing elevated blood pressure on cerebral circulation.
Circulation to the brain is protected not only by autoregulation, which maintains cerebral blood flow (CBF) in spite of a decrease in mean arterial pressure, but also by baroreflexes that tend to prevent or minimize hypotension. Hypertensive patients have impaired cerebral autoregulation and depressed baroreflex sensitivity, suggesting that the brain might be vulnerable to a reduction in blood pressure. Nevertheless, in clinical experience, studies of cerebral blood flow during drug-induced reduction of elevated blood pressure have shown that control of hypertension can usually be achieved without jeopardizing cerebral blood flow or inducing signs and symptoms of cerebral ischemia, even in patients with cerebrovascular disease. Indeed, control of hypertension reduces both the risk of stroke and the recurrence rate for second strokes.


Inference about the expected performance of a data-driven dynamic treatment regime.
A dynamic treatment regime (DTR) comprises a sequence of decision rules, one per stage of intervention, that recommends how to individualize treatment to patients based on evolving treatment and covariate history. These regimes are useful for managing chronic disorders, and fit into the larger paradigm of personalized medicine. The Value of a DTR is the expected outcome when the DTR is used to assign treatments to a population of interest. The Value of a data-driven DTR, estimated using data from a Sequential Multiple Assignment Randomized Trial, is both a data-dependent parameter and a non-smooth function of the underlying generative distribution. These features introduce additional variability that is not accounted for by standard methods for conducting statistical inference, for example, the bootstrap or normal approximations, if applied without adjustment. Our purpose is to provide a feasible method for constructing valid confidence intervals (CIs) for this quantity of practical interest. We propose a conceptually simple and computationally feasible method for constructing valid CIs for the Value of an estimated DTR based on subsampling. The method is self-tuning by virtue of an approach called the double bootstrap. We demonstrate the proposed method using a series of simulated experiments. The proposed method offers considerable improvement in terms of coverage rates of the CIs over the standard bootstrap approach. In this article, we have restricted our attention to Q-learning for estimating the optimal DTR. However, other methods can be employed for this purpose; to keep the discussion focused, we have not explored these alternatives. Subsampling-based CIs provide much better performance compared to standard bootstrap for the Value of an estimated DTR.


RAG2's acidic hinge restricts repair-pathway choice and promotes genomic stability.
V(D)J recombination-associated DNA double-strand breaks (DSBs) are normally repaired by the high-fidelity classical nonhomologous end-joining (cNHEJ) machinery. Previous studies implicated the recombination-activating gene (RAG)/DNA postcleavage complex (PCC) in regulating pathway choice by preventing access to inappropriate repair mechanisms such as homologous recombination (HR) and alternative NHEJ (aNHEJ). Here, we report that RAG2's "acidic hinge," previously of unknown function, is critical for several key steps. Mutations that reduce the hinge's negative charge destabilize the PCC, disrupt pathway choice, permit repair of RAG-mediated DSBs by the translocation-prone aNHEJ machinery, and reduce genomic stability in developing lymphocytes. Structural predictions and experimental results support our hypothesis that reduced flexibility of the hinge underlies these outcomes. Furthermore, sequence variants present in the human population reduce the hinge's negative charge, permit aNHEJ, and diminish genomic integrity.


Photo and hormonal control of meristem identity in the Arabidopsis flower mutants apetala2 and apetala1.
We have analyzed the contributions of phytochrome and gibberellin signal transduction to the control of flower meristem identity in the Arabidopsis mutants apetala1 (ap1) and apetala2 (ap2). ap1 flowers are partially defective for the establishment of flower meristem identity and are characterized by the production of ectopic secondary or axillary flowers and by branching. Axillary flower production is also induced in ap2-1 flowers by short-day photoperiod and is suppressed by hy1, a mutation blocking phytochrome activity. The production of axillary flower by ap2-1 is also suppressed by exogenous gibberellins and by spindly (spy), a mutation that activates basal gibberellin signal transduction in hormone-independent manner. Ectopic axillary flower production and floral branching by ap1 flowers are also suppressed by spy. We conclude that gibberellins promote flower meristem identity and that the inflorescence-like traits of ap2-1 and ap1-1 flowers are due in part to SPY gene activity.


Effects of exercise on aortic input impedance and pressure wave forms in normal humans.
The effects of supine bicycle exercise on the input impedance of the ascending aorta were studied in thirteen male subjects undergoing cardiac catheterization, but in whom no cardiovascular disease was found. Ascending aortic flow velocity and pressure were recorded simultaneously from a multisensor catheter with an electromagnetic flow velocity probe and a pressure sensor mounted at the same location. A second pressure sensor at the catheter tip provided left ventricular pressure. Fick cardiac outputs were used to scale the velocity signal to instantaneous volumetric flow. Input impedance was calculated from 10 harmonics of aortic pressure and flow. For each subject, impedance moduli and phases from a minimum of 15 beats during rest and exercise were averaged. Peripheral resistance decreased from a resting value of 1142 +/- 51 (+/- SE) dynes sec/cm5 to 712 +/- 39 dynes sec/cm5 during exercise. Characteristic impedance remained unchanged with a resting value of 47 +/- 4 dynes sec/cm5 and an exercise value of 45 +/- 4 dynes sec/cm5. These results were associated with an increase in aortic pressure (96 +/- 2 to 111 +/- 2 mm Hg) and pulse wave velocity implying a decrease in aortic compliance. An increase in aortic cross-sectional area apparently offsets the effects of these changes in compliance and pulse wave velocity to result in an unchanged characteristic impedance. The increase in pulse wave velocity caused wave reflections to occur earlier during exercise, but the general characteristics of the pressure wave shapes remained unchanged.


Dynamics of proteins confined in non-ionic bicontinuous microemulsions: a FCS study.
In our present work we present an approach which allows one to confine proteins in structurally nearly identical bicontinuous microemulsions with systematically decreasing water domain size. It is shown that sub-diffusive behaviour occurs already at water domain sizes below 13 nm. However, above 13 nm normal diffusion is seen. Moreover, we compare protein diffusion in microemulsions to the transport of a much smaller fluorescent dye.


Sense of agency and mentalizing: dissociation of subdomains of social cognition in patients with schizophrenia.
The sense of agency, i.e., the sense that "I am the one who is causing an action", and mentalizing, the ability to understand the mental states of other individuals, are key domains of social cognition. It has been hypothesized that an intact sense of agency is an important precondition for higher-level mentalizing abilities. A substantial body of evidence shows that both processes rely on similar brain areas and are severely impaired in schizophrenia, suggesting a close link between agency and mentalizing. Yet this relationship has not been explicitly tested. We investigated 40 individuals with schizophrenia and 40 healthy controls on an agency and mentalizing task. On the agency task, participants carried out simple mouse movements and judged the partially manipulated visual feedback as either self- or other-generated. On the mentalizing task, participants inferred mental states from pictures that depicted others' eyes ("Reading the mind in the eyes test"). Neuropsychological, psychopathological and social functioning levels were also evaluated. Both sense of agency and mentalizing were impaired in schizophrenia patients compared to healthy controls. However, testing for a relationship revealed no significant correlations between the two processes, either in the schizophrenia or the control group. The present findings demonstrate a dissociation of agency and mentalizing deficits in schizophrenia, suggesting that the multifaceted construct of social cognition consists of independent subdomains in healthy and psychiatrically ill individuals.


External quality assessment program for Chlamydia trachomatis diagnostic testing by nucleic acid amplification assays.
We report the results from 57 Australian diagnostic laboratories testing two external quality assessment panels using either the Roche Amplicor Chlamydia trachomatis test (R-PCR) or the Abbott LCx Chlamydia trachomatis assay (A-ligase chain reaction [LCR]). Panel samples were either normal urine spiked with Chlamydia trachomatis antigen or clinical urine specimens. There was no significant difference between laboratories or between assays in detection of C. trachomatis-positive clinical samples. Only at the lower limit of detection of the assays did the R-PCR demonstrate increased sensitivity over the A-LCR in the detection of C. trachomatis antigen. However, it was found that single-sample testing could lead to decreased test sensitivity. Detection of the presence of inhibitors of nucleic acid amplification differed between laboratories.


Running energetics in the pronghorn antelope.
The pronghorn antelope (Antilocapra americana) has an alleged top speed of 100 km h-1, second only to the cheetah (Acionyx jubatus) among land vertebrates, a possible response to predation in the exposed habitat of the North American prairie. Unlike cheetahs, however, pronghorn antelope are distance runners rather than sprinters, and can run 11 km in 10 min, an average speed of 65 km h-1. We measured maximum oxygen uptake in pronghorn antelope to distinguish between two potential explanations for this ability: either they have evolved a uniquely high muscular efficiency (low cost of transport) or they can supply oxygen to the muscles at unusually high levels. Because the cost of transport (energy per unit distance covered per unit body mass) varies as a predictable function of body mass among terrestrial vertebrates, we can calculate the predicted cost to maintain speeds of 65 and 100 km h-1 in an average 32-kg animal. The resulting range of predicted values, 3.2-5.1 ml O2 kg-1 s-1, far surpasses the predicted maximum aerobic capacity of a 32-kg mammal (1.5 ml O2 kg-1 s-1). We conclude that their performance is achieved by an extraordinary capacity to consume and process enough oxygen to support a predicted running speed greater than 20 ms-1 (70 km h-1), attained without unique respiratory-system structures.


Two types of visual dysfunction in autosomal dominant retinitis pigmentosa.
Visual thresholds and rhodopsin levels were determined in nine subjects with autosomal dominant retinitis pigmentosa. The subjects fell into two groups, corresponding to two subtypes of the disease revealed by two-color, dark-adapted static perimetry. In the first of these subtypes, rod-mediated function was variably reduced and was accompanied by a corresponding reduction in cone function in the same retinal region. Dark-adapted threshold elevations varied in a way consistent with decreased quantal absorption by the rods as a result of reduced rhodopsin levels. In the second subtype, rod function was greatly reduced or absent throughout the retina, while cone function was much less severely affected. Although the levels of rhodopsin were only about half of normal, they were much too great to account for the visual threshold elevations on the basis of decreased probabilities of absorption by the visual pigment. Rhodopsin regeneration appeared to follow normal kinetics in patients from both groups. The results indicate that the examples of the two psychophysical subtypes of AD RP investigated here have very different disease manifestations.


Developing the evidence base for gender- and age-relevant school sex education: questionnaire findings from an adolescent sample using an augmented theory of planned behaviour.
Positive adolescent sexual health is supported by effective school-based sex education. Methods to promote positive sexual health need to reflect determinants of contraception intention, which must include understanding gender and age (year group) differences. To date, there has been limited theory-based exploration of these determinants in school age participants, placing limitations on sexual health educators to tailor learning most effectively. Cross-sectional survey data were collected from UK school pupils (n=1378) aged 12-16 years. Measures included theory of planned behaviour, prototype willingness, anticipated regret and knowledge items. Linear regression determined significant predictors of intention to use condoms, the oral contraceptive pill and emergency contraception (EC). The significance of differences by gender and school year was evaluated using t-tests and analysis of variance (ANOVA). Three distinct predictive models emerged for condom, pill and EC use, predicting 36%, 18% and 23% of variance respectively. Attitude, gender and anticipated regret for unprotected sex significantly predicted intention for all types of contraception (P<0.001). The effects of other explanatory variables differed by contraceptive. Girls scored higher on all variables except condom intention, and intention scores peaked in Year 10. Intention to use condoms, the pill and EC have different predictive profiles, with girls more strongly motivated and Year 10 a crucial stage for intention. Social comparisons and control beliefs exert different effects across contraceptive types, whereas attitudes and anticipated regret are consistently strong influences. The findings suggest clear scope for supporting sexual health and well being through modified school sex education.


Visceral afferent stimulation-evoked changes in the release of peptides into the parabrachial nucleus in vivo.
Previous investigations have demonstrated that the peptides substance P (SP), calcitonin gene-related peptide (CGRP), cholecystokinin (CCK), neurotensin (NT) and somatostatin (SOM) significantly modulate the glutamate-mediated transmission of visceral information through the parabrachial nucleus (PBN) to the ventrobasal thalamus. In addition, we have shown that the staining intensity of SOM, CCK and NT in the PBN decreases significantly following 2 h of vagal stimulation as visualized using immunohistochemistry. As well, the staining intensity of both SP and CGRP in the PBN were shown to increase under similar conditions. The present investigation was done to determine whether the altered peptide staining intensity of these peptides observed following 2 h of vagal stimulation was the result of an altered peptide release from terminals within the PBN. Male Sprague-Dawley rats were anesthetized with sodium thiobutabarbitol and instrumented to record blood pressure and heart rate and for the stimulation of the cervical vagus nerve. A push-pull perfusion cannula was lowered into the region of the PBN for the continuous sampling of extracellular fluid. Radioenzymatic quantification of the perfusates for peptide content revealed that the extracellular fluid concentration of CGRP and SP increased significantly during the 2 h of vagal stimulation. When the vagal stimulation was terminated, the release of both CGRP and SP decreased significantly below prestimulated values for approximately 30 min before returning to prestimulated levels shortly thereafter. In contrast, there was a significant decrease in the release of CCK, SOM and NT into the PBN during the period of vagal stimulation. Extracellular perfusate levels of these peptides returned to normal upon termination of stimulation. These results demonstrate that terminal release of CGRP and SP is significantly increased and terminal release of CCK, SOM and NT is significantly decreased in the PBN during 2 h of vagal stimulation. These results are consistent with our previous finding that the immunohistochemical staining intensity of CGRP and SP is increased while that of CCK, SOM and NT is decreased following vagal stimulation.


Association of chronic renal insufficiency with in-hospital outcomes after percutaneous coronary intervention.
The association of chronic renal insufficiency with outcomes after percutaneous coronary intervention (PCI) in the current era of drug-eluting stents and modern antithrombotic therapy has not been well characterized. We queried the 2007-2011 Nationwide Inpatient Sample databases to identify all patients aged ≥18 years who underwent PCI. Multivariable logistic regression was used to compare in-hospital outcomes among patients with chronic kidney disease (CKD), patients with end-stage renal disease (ESRD), and those without CKD or ESRD. Of 3 187 404 patients who underwent PCI, 89% had no CKD/ESRD; 8.6% had CKD; and 2.4% had ESRD. Compared to patients with no CKD/ESRD, patients with CKD and patients with ESRD had higher in-hospital mortality (1.4% versus 2.7% versus 4.4%, respectively; adjusted odds ratio for CKD 1.15, 95% CI 1.12 to 1.19, P<0.001; adjusted odds ratio for ESRD 2.29, 95% CI 2.19 to 2.40, P<0.001), higher incidence of postprocedure hemorrhage (3.5% versus 5.4% versus 6.0%, respectively; adjusted odds ratio for CKD 1.21, 95% CI 1.18 to 1.23, P<0.001; adjusted odds ratio for ESRD 1.27, 95% CI 1.23 to 1.32, P<0.001), longer average length of stay (2.9 days versus 5.0 days versus 6.4 days, respectively; P<0.001), and higher average total hospital charges ($60 526 versus $77 324 versus $97 102, respectively; P<0.001). Similar results were seen in subgroups of patients undergoing PCI for acute coronary syndrome or stable ischemic heart disease. In patients undergoing PCI, chronic renal insufficiency is associated with higher in-hospital mortality, higher postprocedure hemorrhage, longer average length of stay, and higher average hospital charges.


Structural basis of enantioselective inhibition of cyclooxygenase-1 by S-alpha-substituted indomethacin ethanolamides.
The modification of the nonselective nonsteroidal anti-inflammatory drug, indomethacin, by amidation presents a promising strategy for designing novel cyclooxygenase (COX)-2-selective inhibitors. A series of alpha-substituted indomethacin ethanolamides, which exist as R/S-enantiomeric pairs, provides a means to study the impact of stereochemistry on COX inhibition. Comparative studies revealed that the R- and S-enantiomers of the alpha-substituted analogs inhibit COX-2 with almost equal efficacy, whereas COX-1 is selectively inhibited by the S-enantiomers. Mutagenesis studies have not been able to identify residues that manifest the enantioselectivity in COX-1. In an effort to understand the structural impact of chirality on COX-1 selectivity, the crystal structures of ovine COX-1 in complexes with an enantiomeric pair of these indomethacin ethanolamides were determined at resolutions between 2.75 and 2.85 A. These structures reveal unique, enantiomer-selective interactions within the COX-1 side pocket region that stabilize drug binding and account for the chiral selectivity observed with the (S)-alpha-substituted indomethacin ethanolamides. Kinetic analysis of binding demonstrates that both inhibitors bind quickly utilizing a two-step mechanism. However, the second binding step is readily reversible for the R-enantiomer, whereas for the S-enantiomer, it is not. These studies establish for the first time the structural and kinetic basis of high affinity binding of a neutral inhibitor to COX-1 and demonstrate that the side pocket of COX-1, previously thought to be sterically inaccessible, can serve as a binding pocket for inhibitor association.


Proliferative response of corneal endothelial cells from young and older donors.
To compare the effect of epidermal growth factor (EGF), nerve growth factor (NGF), platelet-derived growth factor-BB (PDGF-BB), bovine pituitary extract, and fetal bovine serum (FBS), alone or in combination, on proliferation of human corneal endothelial cells (HCEC) cultured from young (<30 years old) and older donors (>50 years old). Corneas from donors 2 to 79 years old were obtained from the National Disease Research Interchange. Descemet's membrane with intact endothelium was dissected. Cells were isolated by EDTA treatment and cultured to confluence. The HCEC marker, antibody 9.3.E, tested for pure endothelial populations. Antibody Ki67 and ZO-1 tested either before or after cultured cells reached confluence to indicate cell proliferation and cell-cell contact formation. Cell morphology was documented by inverted phase-contrast microscopy. Passages I through VII were used to test the effect of various factors on cell proliferation. For each study, equal numbers of cells were seeded, maintained overnight in 4% FBS to permit cell attachment, washed, and incubated for up to 3 weeks in one of the following: modified Eagle's Minimum Essential Medium (Opti-MEM-I) alone; Opti-MEM-I plus EGF, NGF, PDGF-BB, bovine pituitary extract, or FBS; or a combination of factors. At various times after seeding, cell numbers were determined by electronic cell counter. For each condition, three separate wells were tested and each sample was counted three times. Studies were repeated at least twice using cells from different donors and age groups. Within each study, a one-way ANOVA test was performed to analyze statistical significance. Cells stained positively with antibody 9.3.E, indicating isolation of HCEC and lack of contamination with epithelial cells or keratocytes. Positive staining of Ki67, indicating cycling cells, was found in subconfluent cultures. Plasma membrane-associated ZO-1 staining and lack of Ki67 staining indicated that cultured cells formed a contact-inhibited monolayer. Cultured cells decreased in density, increased in size, and became more heterogeneous depending on donor age and on the number of passages. Incubation in OptiMEM-I promoted attachment and induced a moderate proliferative response above that of MEM (P < 0.001). In general, proliferative responses to growth stimuli were relatively slow, with cell counts generally plateauing 10 to 14 days after exposure to growth-promoting agents. EGF yielded a broad, dose-dependent effect and, at 5-50 ng/mL, peak cell counts were significantly higher (P < 0.001) than basal levels. EGF consistently stimulated proliferation in cells from younger donors, but was less effective in stimulating growth of cells from older donors. NGF did not show a consistent significant stimulatory effect at any concentration tested. PDGF-BB (25 ng/mL) tended to stimulate growth to a greater extent than EGF (P < 0.05) in cultures from the same donor. Pituitary extract significantly increased counts at 1.0 (P < 0.05) to 100 ug/mL (P < 0.001). PDGF-BB plus pituitary extract demonstrated greater stimulation than pituitary extract (P < 0.01) or PDGF-BB alone (P < 0.01). FBS (1%-8%) increased cell numbers in a dose-dependent manner, and, at 4%-8%, yielded counts significantly higher (P < 0.001) than that of any single growth-promoting agent tested. HCEC from both young and older donors can proliferate in vitro in response to growth-promoting agents. Proliferation in the presence of multiple mitogens ceased when confluence was reached, indicating the formation of a contact-inhibited monolayer. In general, cells cultured from young donors were more responsive to the agents tested, but the relative response of HCEC to these agents was similar, regardless of donor age. The relative difference in the extent of the response of the same cell population to different mitogens suggests that these mitogens induce different downstream signals. The relatively robust proliferative response of HCEC to FBS may involve stimulation of multiple downstream signaling pathways may involve stimulation of multiple downstream signaling pathways and/or induce more sustained downstream signaling than the other growth-promoting agents tested.


A novel therapeutic approach to treating obesity through modulation of TGFβ signaling.
Obesity results from disproportionately high energy intake relative to energy expenditure. Many therapeutic strategies have focused on the intake side of the equation, including pharmaceutical targeting of appetite and digestion. An alternative approach is to increase energy expenditure through physical activity or adaptive thermogenesis. A pharmacological way to increase muscle mass and hence exercise capacity is through inhibition of the activin receptor type IIB (ActRIIB). Muscle mass and strength is regulated, at least in part, by growth factors that signal via ActRIIB. Administration of a soluble ActRIIB protein comprised of a form of the extracellular domain of ActRIIB fused to a human Fc (ActRIIB-Fc) results in a substantial muscle mass increase in normal mice. However, ActRIIB is also present on and mediates the action of growth factors in adipose tissue, although the function of this system is poorly understood. In the current study, we report the effect of ActRIIB-Fc to suppress diet-induced obesity and linked metabolic dysfunctions in mice fed a high-fat diet. ActRIIB-Fc induced a brown fat-like thermogenic gene program in epididymal white fat, as shown by robustly increased expression of the thermogenic genes uncoupling protein 1 and peroxisomal proliferator-activated receptor-γ coactivator 1α. Finally, we identified multiple ligands capable of reducing thermogenesis that represent likely target ligands for the ActRIIB-Fc effects on the white fat depots. These data demonstrate that novel therapeutic ActRIIB-Fc improves obesity and obesity-linked metabolic disease by both increasing skeletal muscle mass and by inducing a gene program of thermogenesis in the white adipose tissues.


[A case of acute promyelocytic leukemia (APL) with myeloblastoma in the oral cavity developing after receiving all-trans retinoic acid (ATRA)].
A 44-year-old woman was diagnosed as having acute promyelocytic leukemia (APL) in April 1988. On her first admission, chromosomal translocation (15; 17), +8, and +12 was detected. When she was readmitted to our hospital with the second relapse in May 1990, t(3; 13) and +8 was detected, instead of t(15;17). Complete remission was re-achieved with VP-16, MIT, and BHAC, but the third relapse occurred in September 1990. After obtaining informed consent, she was given etretinate 40 mg per day orally for 17 days, without any effect on leukemia. She was then given all-trans retinoic acid (ATRA) 60 mg per day orally for 29 days. Although a mild granulocytic recovery was observed, no sufficient hematological recovery was obtained (minor response). Besides common side effects of ATRA, such as dry skin and hypertriglycedemia, she had a myeloblastoma in the oral cavity, but it is unknown whether the symptom was a complication of ATRA therapy or not.


Perinatal events and the risk of developing primary sclerosing cholangitis.
To investigate whether perinatal events, intrauterine or postpartum, are associated with the development of primary sclerosing cholangitis (PSC) later in life. Birth records from 97 patients with adult PSC in Sweden were reviewed. Information on perinatal events including medications and complications during pregnancy, gestation length, birth weight and length were collected. Two control children of the same sex were selected for each subject. Conditional multiple logistic regression was used to assess associations of the perinatal measures with development of PSC. No significant associations were found between gestational age, birth length, breastfeeding, and the majority of medical complications including infections or medication during pregnancy for the mothers or postpartum for the children. Vaginal bleeding and peripheral oedema showed associations with PSC, with matched odds ratios of 5.70 (95% CI, 1.13-28.83) and 2.28 (95% CI, 1.04-5.03), respectively. The associations of vaginal bleeding and oedema with subsequent PSC cannot readily be explained, so our findings do not strongly support the hypothesis of a significant role of perinatal events as a risk for the development of PSC later in life.


Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds.
Three dimensional (3D) bioprinting is an emerging technology that enables complex spatial modeling of cell-based tissue engineering products, whose therapeutic potential in regenerative medicine is enormous. However, its success largely depends on the definition of a bioprintable zone, which is specific for each combination of cell-loaded hydrogels (or bioinks) and scaffolds, matching the mechanical and biological characteristics of the target tissue to be repaired. Therefore proper adjustment of the bioink formulation requires a compromise between: (i) the maintenance of cellular critical quality attributes (CQA) within a defined range of specifications to cell component, and (ii) the mechanical characteristics of the printed tissue to biofabricate. Herein, we investigated the advantages of using natural hydrogel-based bioinks to preserve the most relevant CQA in bone tissue regeneration applications, particularly focusing on cell viability and osteogenic potential of multipotent mesenchymal stromal cells (MSCs) displaying tripotency in vitro, and a phenotypic profile of 99.9% CD105+ /CD45,- 10.3% HLA-DR,+ 100.0% CD90,+ and 99.2% CD73+ /CD31- expression. Remarkably, hyaluronic acid, fibrin, and gelatin allowed for optimal recovery of viable cells, while preserving MSC's proliferation capacity and osteogenic potency in vitro. This was achieved by providing a 3D structure with a compression module below 8.8 ± 0.5 kPa, given that higher values resulted in cell loss by mechanical stress. Beyond the biocompatibility of naturally occurring polymers, our results highlight the enhanced protection on CQA exerted by bioinks of natural origin (preferably HA, gelatin, and fibrin) on MSC, bone marrow during the 3D bioprinting process, reducing shear stress and offering structural support for proliferation and osteogenic differentiation.


Immunomodulatory activity of a lupane triterpenoid ester isolated from the eastern Nigeria mistletoe, Loranthus micranthus (Linn).
To provide further evidence for the ethnomedicinal use of the Eastern Nigeria mistletoe, Loranthus micranthus (L. micranthus), as an immunostimulant. Solvent fractions from the crude extract of the mistletoe plant was obtained and screened by the cell mediated delayed type hypersensitivity reaction (DTHR) model in mice. Then the immunomodulatory potentials of a major lupane triterpenoid ester isolated from an active hexane fraction of the Eastern Nigeria mistletoe was investigated. Three lupeol-based triterpenoid esters: 7β 15α-dihydroxyl-lup-20(29)-ene-3β-palmitate (I), 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-stearate (II) and 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-decadecanoate (III) were isolated from the plant leaves epiphyting on a local kola nut tree and were characterized. Compound 1 was subjected to cell proliferation studies using C57Bl/6 splenocytes at three dose levels (5, 25 and 100 μg/mL) in presence of controls. Furthermore, the effect of this compound on IL-8 receptor expression was evaluated at three doses (1, 5 and 10 μg/mL) using the real time polymerase chain reaction assay. This triterpenoid ester produced some enhancement of the splenocytes at the tested doses but at doses higher than 5 μg/mL caused inhibition of the IL-8 receptor expression. The present findings support the ethnomedicinal use of the Eastern Nigeria Mistletoe in the management of diseases affecting the immune system. The triterpenoid(s) have some immunomodulatory abilities on splenocytes and IL-8 receptors and may partly account for the overall immunomodulatory activity of this plant.


Advanced paternal age and childhood cancer in offspring: A nationwide register-based cohort study.
Cancer initiation is presumed to occur in utero for many childhood cancers and it has been hypothesized that advanced paternal age may have an impact due to the increasing number of mutations in the sperm DNA with increasing paternal age. We examined the association between paternal age and specific types of childhood cancer in offspring in a large nationwide cohort of 1,904,363 children born in Denmark from 1978 through 2010. The children were identified in the Danish Medical Birth Registry and were linked to information from other national registers, including the Danish Cancer Registry. In total, 3,492 children were diagnosed with cancer before the age of 15 years. The adjusted hazard ratio of childhood cancer according to paternal age was estimated using Cox proportional hazards regressions. We found a 13% (95% confidence interval: 4-23%) higher hazard rate for every 5 years advantage in paternal age for acute lymphoblastic leukemia, while no clear association was found for acute myeloid leukemia (hazard ratio pr. 5 years = 1.02, 95% confidence interval: 0.80-1.30). The estimates for neoplasms in the central nervous system suggested a lower hazard rate with higher paternal age (hazard ratio pr. 5 years = 0.92, 95% confidence interval: 0.84-1.01). No clear associations were found for the remaining childhood cancer types. The findings suggest that paternal age is moderately associated with a higher rate of childhood acute lymphoblastic leukemia, but not acute myeloid leukemia, in offspring, while no firm conclusions could be made for other specific cancer types.


Visualization of Surface Acoustic Waves in Thin Liquid Films.
We demonstrate that the propagation path of a surface acoustic wave (SAW), excited with an interdigitated transducer (IDT), can be visualized using a thin liquid film dispensed onto a lithium niobate (LiNbO3) substrate. The practical advantages of this visualization method are its rapid and simple implementation, with many potential applications including in characterising acoustic pumping within microfluidic channels. It also enables low-cost characterisation of IDT designs thereby allowing the determination of anisotropy and orientation of the piezoelectric substrate without the requirement for sophisticated and expensive equipment. Here, we show that the optical visibility of the sound path critically depends on the physical properties of the liquid film and identify heptane and methanol as most contrast rich solvents for visualization of SAW. We also provide a detailed theoretical description of this effect.


Helping the Noncompliant Child: An Assessment of Program Costs and Cost-Effectiveness.
Disruptive behavior disorders (DBD) in children can lead to delinquency in adolescence and antisocial behavior in adulthood. Several evidence-based behavioral parent training (BPT) programs have been created to treat early onset DBD. This paper focuses on one such program, Helping the Noncompliant Child (HNC), and provides detailed cost estimates from a recently completed pilot study for the HNC program. The study also assesses the average cost-effectiveness of the HNC program by combining program cost estimates with data on improvements in child participants' disruptive behavior. The cost and effectiveness estimates are based on implementation of HNC with low-income families. Investigators developed a Microsoft Excel-based costing instrument to collect data from therapists on their time spent delivering the HNC program. The instrument was designed using an activity-based costing approach, where each therapist reported program time by family, by date, and for each skill that the family was working to master. Combining labor and non-labor costs, it is estimated that delivering the HNC program costs an average of $501 per family from a payer perspective. It also costs an average of $13 to improve the Eyberg Child Behavior Inventory intensity score by 1 point for children whose families participated in the HNC pilot program. The cost of delivering the HNC program appears to compare favorably with the costs of similar BPT programs. These cost estimates are the first to be collected systematically and prospectively for HNC. Program managers may use these estimates to plan for the resources needed to fully implement HNC.


Mesoscale Mapping of Mouse Cortex Reveals Frequency-Dependent Cycling between Distinct Macroscale Functional Modules.
Connectivity mapping based on resting-state activity in mice has revealed functional motifs of correlated activity. However, the rules by which motifs organize into larger functional modules that lead to hemisphere wide spatial-temporal activity sequences is not clear. We explore cortical activity parcellation in head-fixed, quiet awake GCaMP6 mice from both sexes by using mesoscopic calcium imaging. Spectral decomposition of spontaneous cortical activity revealed the presence of two dominant frequency modes (<1 and ∼3 Hz), each of them associated with a unique spatial signature of cortical macro-parcellation not predicted by classical cytoarchitectonic definitions of cortical areas. Based on assessment of 0.1-1 Hz activity, we define two macro-organizing principles: the first being a rotating polymodal-association pinwheel structure around which activity flows sequentially from visual to barrel then to hindlimb somatosensory; the second principle is correlated activity symmetry planes that exist on many levels within a single domain such as intrahemispheric reflections of sensory and motor cortices. In contrast, higher frequency activity >1 Hz yielded two larger clusters of coactivated areas with an enlarged default mode network-like posterior region. We suggest that the apparent constrained structure for intra-areal cortical activity flow could be exploited in future efforts to normalize activity in diseases of the nervous system.SIGNIFICANCE STATEMENT Increasingly, functional connectivity mapping of spontaneous activity is being used to reveal the organization of the brain. However, because the brain operates across multiple space and time domains a more detailed understanding of this organization is necessary. We used in vivo wide-field calcium imaging of the indicator GCaMP6 in head-fixed, awake mice to characterize the organization of spontaneous cortical activity at different spatiotemporal scales. Correlation analysis defines the presence of two to three superclusters of activity that span traditionally defined functional territories and were frequency dependent. This work helps define the rules for how different cortical areas interact in time and space. We provide a framework necessary for future studies that explore functional reorganization of brain circuits in disease models.


P-O-B(3) linkages in borophosphate glasses evidenced by high field (11)B/(31)P correlation NMR.
The long-standing debate about the presence of P-O-B(3) linkages in glasses has been solved by high-field scalar correlation NMR. Previously suggested by dipolar NMR methods, the presence of such species has been definitively demonstrated by (11)B((31)P) J-HMQC NMR techniques. The results indicate that borophosphate networks contain P-O-B(3) bonds and thus present a higher degree of atomic homogeneity than previously thought.


Investigation of dermal toxicity of ionic liquids in monolayer-cultured skin cells and 3D reconstructed human skin models.
Ionic liquids have gained increasing attention in the chemical industry as potential green substitutes for traditional solvents. However, little is known about toxicity of ionic liquids on the skin, a major exposure portal to toxic substances. Here, we evaluated dermal toxicity of ionic liquids using human keratinocyte and fibroblast cell line, 3D reconstructed human epidermis, and full-thickness model to investigate underlying mechanisms. Cytotoxicity of ionic liquids was evaluated for representative anions, [TFSI], [PF6], [BF4], and [DCA], as well as for cations, [EMIM], [BMPY], [TBA] and [Zn], in human keratinocyte cell line, HaCaT, and human dermal fibroblasts. In our results, significant cytotoxicity was induced by ionic liquids with [TFSI] in both cell lines. Notably, cytotoxicity of [TFSI] containing ionic liquids was comparable to xylene, a toxic conventional organic solvent. Fluorescent and flow cytometric analysis revealed that [TFSI]-exposed cells underwent necrotic cell death. Reactive oxygen species (ROS) was increased while the amount of glutathione was decreased by [TFSI] in dose-dependent manner, which was reversed by antioxidant, N-acetylcysteine. In 3D reconstructed human epidermis and full-thickness model, a single application of [TFSI] induced toxicity although it was minimal and largely limited to epidermal layer. Collectively, these results demonstrated potential dermal toxicity of ionic liquids.


Female parental consanguinity is associated with a reduced ovarian reserve.
Is parental consanguinity associated with a reduced ovarian reserve in women from the Arabian Peninsula, comparing anti-Müllerian hormone (AMH) and antral follicle count (AFC)? Retrospective large-scale observational study including 2482 women from the Arabian Peninsula, aged 19-49 years, who had their serum AMH and AFC measured as part of their fertility assessment, from May 2015 to November 2019. Consanguinity was defined as women whose parents were first-degree or second-degree cousins. Serum AMH was measured for all participants. A total of 2198 women were included: 605 in the consanguine group (27.53%), 1593 (72.47%) in the non-consanguine group. There were no significant differences between groups in terms of body mass index, years of infertility or smoking status. Women from the consanguine group were significantly younger (mean age 33.74 ± 6.64 years) compared with the non-consanguine group (mean age 34.78 ± 6.64 years, P < 0.0001). Median AMH and AFC for the consanguine group were 1.90 ng/ml (min-max: 0.01-23.8) and 11 (0-80), respectively, and for the non-consanguine group 1.84 ng/ml (min-max: 0.01-23.0) and 11 (0-60), respectively. AMH and AFC exhibit an age-dependent decline. As both parameters are age-dependent, the multivariate analysis showed that women from the consanguine group presented significantly lower AMH (coefficient of variation [CV] -0.07 ± 0.03, P = 0.036) and AFC (CV -0.16 ± 0.06, P = 0.003) compared with non-consanguine women, and the highest differences were found for women below 35 years of age (AMH median [min-max]: 2.82 ng/ml (0.01-23.80) versus 2.92 ng/ml (0.01-23.00); P = 0.035; AFC median [min-max]: 15 (0-80) versus 14 (0-80); P = 0.001). The adjusted analysis by age indicates that female parental consanguinity is associated with reduced ovarian reserve in the studied population. Clinical evaluation should include extensive family history and subsequent counselling of the affected couples.


Insights into genome plasticity and pathogenicity of the plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria revealed by the complete genome sequence.
The gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria is the causative agent of bacterial spot disease in pepper and tomato plants, which leads to economically important yield losses. This pathosystem has become a well-established model for studying bacterial infection strategies. Here, we present the whole-genome sequence of the pepper-pathogenic Xanthomonas campestris pv. vesicatoria strain 85-10, which comprises a 5.17-Mb circular chromosome and four plasmids. The genome has a high G+C content (64.75%) and signatures of extensive genome plasticity. Whole-genome comparisons revealed a gene order similar to both Xanthomonas axonopodis pv. citri and Xanthomonas campestris pv. campestris and a structure completely different from Xanthomonas oryzae pv. oryzae. A total of 548 coding sequences (12.2%) are unique to X. campestris pv. vesicatoria. In addition to a type III secretion system, which is essential for pathogenicity, the genome of strain 85-10 encodes all other types of protein secretion systems described so far in gram-negative bacteria. Remarkably, one of the putative type IV secretion systems encoded on the largest plasmid is similar to the Icm/Dot systems of the human pathogens Legionella pneumophila and Coxiella burnetii. Comparisons with other completely sequenced plant pathogens predicted six novel type III effector proteins and several other virulence factors, including adhesins, cell wall-degrading enzymes, and extracellular polysaccharides.


Patterns of alcohol use, their correlates, and impact in male drinkers: a population-based survey from Goa, India.
Associations between low socio-economic class and alcohol use disorders are relatively well established in developed countries; however, there is comparably little research in India and other developing countries on the associations between socio-economic class, drinking patterns, and alcohol-related problems. We sought to assess drinking patterns and adverse outcomes among male drinkers and examine whether the association between drinking patterns and adverse outcomes differ by socioeconomic class. Population survey of 732 male drinkers screened from 1,899 men, aged 18 to 49 years, randomly selected from rural and urban communities in northern Goa, India. Usual quantity of alcohol consumed by 14.8% (rural 16.8%; urban 13.6%) current drinkers is at high-risk level. About 28.6% (rural 31%; urban 27.2%) and 33.7% (rural 30.5%; urban 35.5%) of current drinkers reported monthly or more frequent heavy episodic drinking and drunkenness, respectively. Lower education and lower standard of living (SLI) were associated with higher usual quantity of alcohol consumption. More frequent heavy episodic drinking was associated with older age, being separated, lower education, and lower standard of living; weekly or more frequent drunkenness was associated only with rural residence. All three risky drinking patterns were associated with common mental disorders, sexual risk, intimate partner violence, acute alcohol-related consequences, and alcohol dependence. Significant interactions between SLI and risky alcohol use patterns suggested an increased risk of intimate partner violence among men with risky drinking and lower SLI. Risky drinking patterns are common among male drinkers in Goa and associated with lower socio-economic class. A range of adverse health and social outcomes were associated with risky drinking across all socio-economic classes. Alcohol policy should target risky drinking patterns, particularly among poorer men, to reduce the health and social burden of alcohol use in India.


Serology and eosinophil count in the diagnosis and management of strongyloidiasis in a non-endemic area.
Strongyloidiasis is a chronic infection that may result in significant morbidity; however, diagnosis and management remain problematic. The objective of this study was to 1) evaluate the demographic, clinical, and laboratory features of 76 consecutive individuals who had Strongyloides stercoralis larvae identified in their fecal specimens; 2) determine the sensitivity of the Centers for Disease Control and Prevention (CDC) enzyme immunoassay (EIA) for detecting antibodies to Strongyloides in those with confirmed infection; and 3) assess the serologic responses and changes in eosinophil counts following treatment. Most (96%) cases occurred in immigrants, but some patients had immigrated as long as 40 years earlier. The CDC Strongyloides EIA had a sensitivity of 94.6% (95% confidence interval = 92.0-97.2%) in this patient population with proven infection. Serologic and eosinophil counts decreased after therapy, suggesting that they may be useful markers of treatment success.


The protective nature of Chironomus luridus larval tubes against copper sulfate.
The objective of this study was to determine whether the tubes in which Chironomus larvae dwell protect them against chemical toxicants. A laboratory culture of an Israeli benthic midge, Chironomus luridus, was exposed to copper sulfate. Two conditions were tested in bioassay experiments: larvae within silt tubes and larvae without tubes. The non toxic, anionic, fluorescent dye, fluorescein, was used to examine the effect of sub-lethal copper sulfate concentrations on the permeability of cuticular, gill and gut epithelia of the chironomids. Increased cell permeability, which is the cause of cell damage, was reflected by an increase in fluorescence intensity. Following exposure to copper sulfate, higher fluorescence was found in different body compartments: midgut, hindgut, tracheal gills, fat body and muscles, and the Malpighian tubules. The effect was significantly higher in tube-free larvae when compared to silt tube dwelling larvae. We conclude that in addition to its other functions in feeding, respiration, and anti-predation shelter, the Chironomus luridus tube protects its inhabitant from toxins such as copper sulfate.


Verbal mediation of cognition in children with specific language impairment.
Private speech (PS) and inner speech (IS) are thought to be functionally important for children's and adults' cognition, but they have not been studied systematically in children with specific language impairment (SLI). Participants were 21 children with SLI (7-11 years, expressive or receptive verbal IQ ≤ 75, nonverbal IQ ≥ 84) and 21 age- and nonverbal IQ-matched controls. Participants completed three sets of Tower of London problems: one with no dual task (PS condition), one with articulatory suppression, and one while foot tapping (control condition). Participants also completed a digit span task. There was no group difference in the susceptibility of Tower of London performance to articulatory suppression, but the PS of the SLI group was less internalized than that of the controls on both tasks. The findings suggest that children with SLI experience a significant delay in the development of PS/IS, but that their PS/IS is effective for Tower of London performance in middle childhood. Findings are discussed with reference to the interpretation of the nonlinguistic deficits associated with SLI, and in terms of clinical implications.


New sunphotometer for network operation.
A handheld sunphotometer has been developed and is described. It operates for 9 wavelengths in the visible and near infrared. The sensitivity range is rather broad, so it can be used in turbid as well as clear atmospheres. The operation is simple, and a digital readout helps prevent operator biases. The instrument is sun powered and suitable for network operations. Determination of the atmospheric turbidity due to aerosol and the column content of O3, NO2, H2O is possible.


Regulation of CD8+ T-cell cytotoxicity in HIV-1 infection.
Understanding the mechanisms involved in cellular immune responses against control of human immunodeficiency virus (HIV) infection is key to development of effective immunotherapeutic strategies against viral proliferation. Clear insights into the regulation of cytotoxic CD8+ T cells is crucial to development of effective immunotherapeutic strategies due to their unique ability to eliminate virus-infected cells during the course of infection. Here, we reviewed the roles of transcription factors, co-inhibitory molecules and regulatory cytokines following HIV infection and their potential significance in regulating the cytotoxic potentials of CD8+ T cells.


Fluid biomarker and electrophysiological outcome measures for progressive MS trials.
Progressive multiple sclerosis (MS) is characterized by insidious clinical worsening that is difficult to accurately quantify and predict. Biofluid markers and electrophysiological measures are potential candidate outcome measures in clinical trials, allowing the quantification of nervous damage occurring in the disease. Neurofilaments are highly specific neuronal proteins. They may have come closest to such applications by their higher concentrations repeatedly demonstrated in cerebrospinal fluid (CSF) in all stages of MS, during relapses, their responsiveness to disease-modifying treatments in relapsing and progressive MS and their associations with measures of inflammatory and degenerative magnetic resonance imaging (MRI) outcomes. Digital single-molecule array (Simoa) technology improves accuracy of bioassays in the quantification of neurofilament light chain (NfL) in serum and plasma. NfL seems to mark a common final path of neuroaxonal injury independent of specific causal pathways. CSF and blood levels of NfL are highly correlated across various diseases including MS, suggesting that blood measurements may be useful in assessing response to treatment and predicting future disease activity. Other biomarkers like matrix metalloproteinases, chemokines, or neurotrophic factors have not been studied to a similar extent. Such measures, especially in blood, need further validation to enter the trial arena or clinical practice. The broadening armamentarium of highly sensitive assay technologies in the future may shed even more light on patient heterogeneity and mechanisms leading to disability in MS. Evoked potentials (EPs) are used in clinical practice to measure central conduction of central sensorimotor pathways. They correlate with and predict the severity of clinical involvement of their corresponding function. Their validation for use in multicenter studies is still lacking, with the exception of visual EPs. If further validated, EPs and fluid biomarkers would represent useful outcome measures for clinical trials, being related to specific mechanisms of the ongoing pathologic changes.


Variability of objective and subjective intensities during ball drills in youth soccer players.
The aim of this study was to examine the intrasubject (intraclass coefficient correlation [ICC]) and intersubject variability (coefficient of variation [CV]) of soccer ball drills (BD) involving or not opposition in male youth soccer. For this purpose, a collective ball dribbling (DB) exercise and a 7-a-side ball game without coach encouragements were considered. Exercise intensity was assessed as heart rate (HR), training load (TL), and perceived exertion scales. Fourteen U-14 male soccer players (age, 14.79 ± 0.43 years and experience, 6.5 years) of a Spanish First Division club academy participated in the study. Ball drills were examined for variability over 5 successive training sessions in similar field conditions. Results showed that 7-a-side was significantly (p = 0.000) more demanding than DB. Indeed the TL, HRmax, HRmean, overall perceived exertion, and leg muscular perceived exertion (MPE) resulted 141, 8.7, 11, 56, and 72%, higher in 7-a-side than in DB, respectively. In the 7-a-side condition, good intersubject (CV < 10%) and low intrasubject (ICC < 0.7) variability were observed. In the DB condition, CVs were below 10% CV only for HR variables and the ICC values were higher than 0.7 only for MPE. Despite the moderate reproducibility of BD not considering opponents, this condition did not reveal to induce homogeneous physiological responses in young soccer players. Therefore, the use of this kind of drills may be questionable when considered as alternative of moderate intensity generic aerobic training. Despite the higher interaction between players variability in the opposition drills resulted lower, this suggests their use as a specific conditioning exercise.


Gastrin-releasing peptide (GRP) in the ovine uterus: regulation by interferon tau and progesterone.
Gastrin-releasing peptide (GRP) is abundantly expressed by endometrial glands of the ovine uterus and processed into different bioactive peptides, including GRP1-27, GRP18-27, and a C-terminus, that affect cell proliferation and migration. However, little information is available concerning the hormonal regulation of endometrial GRP and expression of GRP receptors in the ovine endometrium and conceptus. These studies determined the effects of pregnancy, progesterone (P4), interferon tau (IFNT), placental lactogen (CSH1), and growth hormone (GH) on expression of GRP in the endometrium and GRP receptors (GRPR, NMBR, BRS3) in the endometrium, conceptus, and placenta. In pregnant ewes, GRP mRNA and protein were first detected predominantly in endometrial glands after Day 10 and were abundant from Days 18 through 120 of gestation. Treatment with IFNT and progesterone but not CSH1 or GH stimulated GRP expression in the endometrial glands. Western blot analyses identified proGRP in uterine luminal fluid and allantoic fluid from Day 80 unilateral pregnant ewes but not in uterine luminal fluid of either cyclic or early pregnant ewes. GRPR mRNA was very low in the Day 18 conceptus and undetectable in the endometrium and placenta; NMBR and BRS3 mRNAs were undetectable in ovine uteroplacental tissues. Collectively, the present studies validate GRP as a novel IFNT-stimulated gene in the glands of the ovine uterus, revealed that IFNT induction of GRP is dependent on P4, and found that exposure of the ovine uterus to P4 for 20 days induces GRP expression in endometrial glands.