Alcohol-related and alcohol-free activity participation and enjoyment among college students: a behavioral theories of choice analysis.
College student alcohol abuse remains a significant public health problem, and there is a need for theory-driven and empirically based models to guide prevention efforts. Behavioral theories of choice assume that the decision to consume alcohol is influenced by the relative value of alcohol versus other available activities. In the present study, a sample of college student drinkers (N=108; 56% female, 44% male) who had previously completed a mandatory alcohol intervention completed a measure of alcohol-related and alcohol-free activity participation and enjoyment. The goals of the study were to examine the influence of drinking quantity and contextual variables on activity enjoyment and to identify enjoyable alcohol-free activities that take place on evenings when students might otherwise be drinking. Overall, students found alcohol-related activities more enjoyable than alcohol-free activities, and drinking quantity was positively related to enjoyment. However, alcohol-free activities such as watching movies, going to the theater or museums, going to bars or parties, hanging out with friends, eating at restaurants, and engaging in creative activity were generally as enjoyable as drinking. Alcohol-free activities that included peers or dates were more enjoyable than solitary activities. Men were less likely to engage in alcohol-free activities that included peers and reported less enjoyment related to alcohol-free activities than did women. Further research is required to identify procedures for increasing participation in alcohol-free activities and to determine whether increased alcohol-free activity participation results in decreased alcohol consumption.


A Novel Glycolipid Antigen for NKT Cells That Preferentially Induces IFN-γ Production.
In this article, we characterize a novel Ag for invariant NKT (iNKT) cells capable of producing an especially robust Th1 response. This glycosphingolipid, DB06-1, is similar in chemical structure to the well-studied α-galactosylceramide (αGalCer), with the only change being a single atom: the substitution of a carbonyl oxygen with a sulfur atom. Although DB06-1 is not a more effective Ag in vitro, the small chemical change has a marked impact on the ability of this lipid Ag to stimulate iNKT cells in vivo, with increased IFN-γ production at 24 h compared with αGalCer, increased IL-12, and increased activation of NK cells to produce IFN-γ. These changes are correlated with an enhanced ability of DB06-1 to load in the CD1d molecules expressed by dendritic cells in vivo. Moreover, structural studies suggest a tighter fit into the CD1d binding groove by DB06-1 compared with αGalCer. Surprisingly, when iNKT cells previously exposed to DB06-1 are restimulated weeks later, they have greatly increased IL-10 production. Therefore, our data are consistent with a model whereby augmented and or prolonged presentation of a glycolipid Ag leads to increased activation of NK cells and a Th1-skewed immune response, which may result, in part, from enhanced loading into CD1d. Furthermore, our data suggest that strong antigenic stimulation in vivo may lead to the expansion of IL-10-producing iNKT cells, which could counteract the benefits of increased early IFN-γ production.


In vivo electroporation: a powerful and convenient means of nonviral gene transfer to tissues of living animals (Review).
Although in vivo electroporation is currently an unfamiliar nonviral means of gene transfer, accounting for only about 1% of total studies related to in vivo gene transfer and gene therapy, it may be extensively used for experimental and therapeutic purposes in the near future. Like other nonviral methods, in vivo electroporation has a variety of advantages over viral vectors as: any types of cells and tissues in theory could become a target, handling is easy and quickly done within a matter of second, repeated administration of DNA is possible, no immunogenicity is expected, and there is no constraints on amounts and sizes of DNA to be used. Gene transfer efficiency of in vivo electroporation was found to be equivalent to or even superior to that of in vivo lipofection, gene gun and direct DNA injection methods. Although gene expression exerted is transient and foreign genes are likely to be present in an episomal form when transferred by in vivo electroporation, foreign gene products could be detected for more than 1 month depending on tissues and DNA constructs used. Gene expression generated by in vivo electroporation could be controlled to a certain extent in a tissue- or cell-specific manner, and be induced as intended. Perhaps better appraisal of in vivo electroporation as a nonviral gene transfer method should be brought forth in the future after more detailed analyses.


Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa.
Post-translational modification by bonding of small ubiquitin-like modifier (SUMO) peptides influences various cellular functions, and is regulated by SUMO-specific proteases (SENPs). Several proteins have been suggested to have diverse impact on insulin synthesis and secretion through SUMO modification in β cells. However, the role of SUMO modification in β cell mass has not been established. Here, we examined the changes in expression of Senp in INS1 cells and pancreatic islets under diabetes-relevant stress conditions and associated changes in β cell mass. Treatment with 25 mM glucose for 72 h induced Senp2 mRNA expression but not that of Senp1 in INS1 cells. Immunohistochemical staining with anti-SENP2 antibody on human pancreas sections revealed that SENP2 was localized in the nucleus. Moreover, in a patient with type 2 diabetes, SENP2 levels were enhanced, especially in the cytoplasm. Senp2 cytoplasmic levels were also increased in islet cells in obese diabetic mice. Cell number peaked earlier in INS1 cells cultured in high-glucose conditions compared to those cultured in control media. This finding was associated with increased Ccnd1 mRNA expression in high-glucose conditions, and siRNA-mediated Senp2 suppression abrogated it. Mafa expression, unlike Pdx1, was also dependent on Senp2 expression during high-glucose conditions. In conclusion, Senp2 may play a role in β cell mass in response to chronic high-glucose through Cyclin D1 and Mafa.


The prevention of wound infections with the use of local antibiotics.
The local use of kanamycin, a broad spectrum antibiotic, has been successful in preventing primary wound infections. There have been no instances of drug reaction or sensitivity when kanamycin is used as a dilute topical irrigating solution. Secondary wound infections, which occurred in two cases, responded to local and systemic antibiotic therapy. Kanamycin can be recommended as an effective and safe topical antibiotic to prevent wound infections in foot surgery.


Quantifying Evidence for-and against-Granger Causality with Bayes Factors.
Testing for Granger causality relies on estimating the capacity of dynamics in one time series to forecast dynamics in another. The canonical test for such temporal predictive causality is based on fitting multivariate time series models and is cast in the classical null hypothesis testing framework. In this framework, we are limited to rejecting the null hypothesis or failing to reject the null - we can never validly accept the null hypothesis of no Granger causality. This is poorly suited for many common purposes, including evidence integration, feature selection, and other cases where it is useful to express evidence against, rather than for, the existence of an association. Here we derive and implement the Bayes factor for Granger causality in a multilevel modeling framework. This Bayes factor summarizes information in the data in terms of a continuously scaled evidence ratio between the presence of Granger causality and its absence. We also introduce this procedure for the multilevel generalization of Granger causality testing. This facilitates inference when information is scarce or noisy or if we are interested primarily in population-level trends. We illustrate our approach with an application on exploring causal relationships in affect using a daily life study.


Gene expression in learning processes.
It has repeatedly been shown that long-term memory formation involves neuronal gene expression. In this article several different roles for neuronal gene function in a context of learning are considered: maintenance of neural functioning, replenishment of cellular elements that are exhausted in response to massive neuronal stimulation accompanying behavioral training, maintenance of the plastically reorganized neuronal connections, and finally integration of information at the level of transcription factor-promoter interaction. It is strongly advocated that only careful scrutiny of learning-related gene expression phenomena may aid in understanding of the complex learning process.


Altered Neural Networks in the Papez Circuit: Implications for Cognitive Dysfunction after Cerebral Ischemia.
Cerebral ischemia remains a leading cause of mortality worldwide. Although the incidence of death has decreased over the years, surviving patients may suffer from long-term cognitive impairments and have an increased risk for dementia. Unfortunately, research aimed toward developing therapies that can improve cognitive outcomes following cerebral ischemia has proved difficult given the fact that little is known about the underlying processes involved. Nevertheless, mechanisms that disrupt neural network activity may provide valuable insight, since disturbances in both local and global networks in the brain have been associated with deficits in cognition. In this review, we suggest that abnormal neural dynamics within different brain networks may arise from disruptions in synaptic plasticity processes and circuitry after ischemia. This discussion primarily concerns disruptions in local network activity within the hippocampus and other extra-hippocampal components of the Papez circuit, given their role in memory processing. However, impaired synaptic plasticity processes and disruptions in structural and functional connections within the Papez circuit have important implications for alterations within the global network, as well. Although much work is required to establish this relationship, evidence thus far suggests there is a link. If pursued further, findings may lead toward a better understanding of how deficits in cognition arise, not only in cerebral ischemia, but in other neurological diseases as well.


Breaking the cipher: ant eavesdropping on the variational trail pheromone of its termite prey.
Predators may eavesdrop on their prey using innate signals of varying nature. In regards to social prey, most of the prey signals are derived from social communication and may therefore be highly complex. The most efficient predators select signals that provide the highest benefits. Here, we showed the use of eusocial prey signals by the termite-raiding ant Odontoponera transversaO. transversa selected the trail pheromone of termites as kairomone in several species of fungus-growing termites (Termitidae: Macrotermitinae: Odontotermes yunnanensis, Macrotermes yunnanensis, Ancistrotermes dimorphus). The most commonly predated termite, O. yunnanensis, was able to regulate the trail pheromone component ratios during its foraging activity. The ratio of the two trail pheromone compounds was correlated with the number of termites in the foraging party. (3Z)-Dodec-3-en-1-ol (DOE) was the dominant trail pheromone component in the initial foraging stages when fewer termites were present. Once a trail was established, (3Z,6Z)-dodeca-3,6-dien-1-ol (DDE) became the major recruitment component in the trail pheromone and enabled mass recruitment of nest-mates to the food source. Although the ants could perceive both components, they revealed stronger behavioural responses to the recruitment component, DDE, than to the common major component, DOE. In other words, the ants use the trail pheromone information as an indication of suitable prey abundance, and regulate their behavioural responses based on the changing trail pheromone component. The eavesdropping behaviour in ants therefore leads to an arms race between predator and prey where the species specific production of trail pheromones in termites is targeted by predatory ant species.


Addressing the Diversity-Validity Dilemma Using Situational Judgment Tests.
To examine the magnitudes of score differences across different demographic groups for three academic (grade point average [GPA], old Medical College Admission Test [MCAT], and MCAT 2015) and one nonacademic (situational judgment test [SJT]) screening measures and one nonacademic (multiple mini-interview [MMI]) interview measure (analysis 1), and the demographic implications of including an SJT in the screening stage for the pool of applicants who are invited to interview (analysis 2). The authors ran the analyses using data from New York Medical College School of Medicine applicants from the 2015-2016 admissions cycle. For analysis 1, effect sizes (Cohen d) were calculated for GPA, old MCAT, MCAT 2015, CASPer (an online SJT), and MMI. Comparisons were made across gender, race, ethnicity (African American, Hispanic/Latino), and socioeconomic status (SES). For analysis 2, a series of simulations were conducted to estimate the number of underrepresented in medicine (UIM) applicants who would have been invited to interview with different weightings of GPA, MCAT, and CASPer scores. A total of 9,096 applicants were included in analysis 1. Group differences were significantly smaller or reversed for CASPer and MMI compared with the academic assessments (MCAT, GPA) across nearly all demographic variables/indicators. The simulations suggested that a higher weighting of CASPer may help increase gender, racial, and ethnic diversity in the interview pool; results for low-SES applicants were mixed. The inclusion of an SJT in the admissions process has the potential to widen access to medical education for a number of UIM groups.


Use of NH (A3Π-X3Σ-) sonoluminescence for diagnostics of nonequilibrium plasma produced by multibubble cavitation.
In this work, the sonoluminescence of NH radicals has been evaluated as a new spectroscopic probe for the nonequilibrium plasma produced by multibubble cavitation in liquids. The experiments were performed in aqueous ammonia solutions subjected to power ultrasound at low and high frequencies and under two different rare gases (Ar and Xe). Sonoluminescence (SL) spectroscopy focuses on the emission of the two present systems: NH (A3Π-X3Σ-) and OH (A2Σ+-X2Π). Both spectroscopic systems indicate the absence of thermal equilibrium during bubble collapse (Tv > Tr) irrespective of the saturating gas. When Ar is used as the saturating gas, these emissions can be fitted using Specair software and the corresponding rovibronic temperatures are derived. Both species indicate a net increase in vibrational temperatures with the US frequency. In Xe, the SL spectra exhibit OH (C2Σ+-A2Σ+) and NH (c1Π-a1Δ) emission bands indicating a higher electron temperature compared to Ar. However, in Xe, the SL spectra cannot be satisfactorily fitted because of significant line broadening. The estimation of the intrabubble pressure via SL simulation using Specair software is discussed. Monitoring of the sonochemical activity indicates the formation of H2 and N2H4, while no H2O2 accumulates under these conditions. In the presence of Xe, NO is also formed as a sonolysis product. The appearance of new possible reaction pathways under Xe is made possible by the higher plasma electron density and correlates with SL data.


Collective dynamics of red blood cells on an in vitro microfluidic platform.
Understanding the dynamics of blood flow in physiologically relevant confinements turns out to be an outstanding proposition in biomedical research. Despite the large number of studies being reported to theoretically elucidate the dynamics of red blood cells (RBCs) in confined geometries, in vitro experimental studies unveiling the implications of the collective dynamics of red blood cells in physiologically relevant bio-mimetic microfluidic channels remain elusive. Here, we investigate the implications of complex dynamvic interactions between the whole blood and a deformable channel wall fabricated using a hydrogel matrix. For a range of flow rates, we map the trajectories of the RBCs for varying levels of softness of the microchannel wall. We compare these scenarios with the reference cases of rigid polydimethylsiloxane (PDMS) channels. Our results reveal that the smallest channels investigated herein exhibit the most intricate interactions between the collective dynamics of the RBC and the wall flexibility, attributable to confinement-induced hydrodynamic interactions in the presence of spatially varying shear rates. These results may open up new paradigms in conceptual understanding of in vivo dynamics of blood flow through simple in vitro experiments on a simple microfluidic platform.


Short communication: Molecular epidemiology, phylogeny, and phylodynamics of CRF63_02A1, a recently originated HIV-1 circulating recombinant form spreading in Siberia.
The HIV-1 epidemic in Russia is dominated by the former Soviet Union subtype A (A(FSU)) variant, but other genetic forms are circulating in the country. One is the recently described CRF63_02A1, derived from recombination between a CRF02_AG variant circulating in Central Asia and A(FSU), which has spread in the Novosibirsk region, Siberia. Here we phylogenetically analyze pol and env segments from 24 HIV-1 samples from the Novosibirsk region collected in 2013, with characterization of three new near full-length genome CRF63_02A1 sequences, and estimate the time of the most recent common ancestor (tMRCA) and the demographic growth of CRF63_02A1 using a Bayesian method. The analyses revealed that CRF63_02A1 is highly predominant in the Novosibirsk region (81.2% in pol sequences) and is transmitted both among injecting drug users and by heterosexual contact. Similarity searches with database sequences combined with phylogenetic analyses show that CRF63_02A1 is circulating in East Kazakhstan and the Eastern area of Russia bordering China. The analyses of near full-length genome sequences show that its mosaic structure is more complex than reported, with 18 breakpoints. The tMRCA of CRF63_02A1 was estimated around 2006, with exponential growth in 2008-2009 and subsequent stabilization. These results provide new insights into the molecular epidemiology, phylogeny, and phylodynamics of CRF63_02A1.


Supplementation with fish oil and coconut fat prevents prenatal stress-induced changes in early postnatal development.
Adequate development of the central nervous system depends on prenatal and postnatal factors. On one hand, prenatal stress (PNS) has been implicated in impaired development of the offspring. On other hand, nutritional factors during pregnancy and lactation can influence fetal and postnatal growth. This study assessed the postnatal development of rat offspring exposed to PNS, which consisted of restraint and bright lights, 3 times/day, from days 14 to 20 of pregnancy, whose mothers were fed different diets during pregnancy and lactation: regular diet, diet supplemented with coconut fat or fish oil. When pregnancy was confirmed, they were distributed into control (CTL) or PNS groups. At birth, PNS males and females weighed less than those in the group CTL. At 21 days of age, this alteration was no longer observed with fish oil and coconut fat groups. PNS and coconut fat diet induced increased locomotor activity in 13 day old male and female pups, and this effect was prevented by fish oil supplementation only in females. In conclusion, postnatal development from birth to weaning was influenced by PNS and diet and some of those alterations were prevented by coconut fat and fish oil.


Long-term safety of infliximab for the treatment of inflammatory bowel disease: a single-centre cohort study.
This study evaluates the long-term safety of infliximab in patients with inflammatory bowel disease (IBD) treated with the drug over a 14-year period. The medical records of 734 patients with IBD treated with infliximab and 666 control patients not treated with infliximab were reviewed for adverse events. The time of onset and outcome, severity and concomitant medication were recorded. Patients and controls were followed up for serious adverse events for a median time of 58 months (IQR 33-88) and 144 months (IQR 83-163), respectively. 112 severe adverse events occurred in 93 patients (13%) treated with infliximab and 157 occurred in 126 (19%) control patients (OR 1.33 (95% CI 0.56 to 3.00, p = 0.45). There was no difference between the two groups in mortality, malignancies and infection rate. Tuberculosis was diagnosed in two patients receiving infliximab who had negative skin tests at baseline whereas none of 16 patients with positive skin tests who received prophylaxis developed tuberculosis. Concomitant treatment with steroids was the only independent risk factor for infections in patients treated with infliximab (OR 2.69 (95% CI 1.18 to 6.12), p = 0.018). The most commonly observed systemic side effects were skin eruptions including psoriasiform eruptions in 150 patients (20%). Long-term infliximab treatment had a good overall safety profile in the patient cohort studied.


Protein kinase activator 1-oleoyl-2-acetyl-sn-glycerol inhibits two types of calcium currents in GH3 cells.
Two types of Ca2+ currents, high-threshold, long-lasting, or L currents and low-threshold, transient, or T currents, are present in many excitable cells. L-type Ca2+ current is modulated by, among others, beta- and alpha-adrenoreceptors and intracellular Ca2+, but modulation of T-type Ca2+ current is less well established. 1-Oleoyl-2-acetyl-sn-glycerol (OAG), a synthetic activator of protein kinase C (PKC), modulates whole cell Ca2+ currents in a variety of excitable cells. Whether activators of PKC affect preferentially L and T types of Ca2+ currents is unknown. We tested OAGs effects on whole cell Ca2+ currents in the clonal GH3 line of anterior pituitary cells. The currents were measured using the whole cell patch-clamp method. Four to 60 microM OAG reversibly reduced Ca2+ currents produced by test potentials to 10 mV, and the inhibition was half maximal at approximately 25 microM. Such concentrations depress Ca2+ currents in chick embryo dorsal root ganglion (DRG) cells and clonal AtT-20 pituitary cells. To test whether OAG acted preferentially on L or T current, we separated the two using depolarizing prepulses to inactivate T current. OAG (40 microM) attenuated T currents by 60% and L currents by 50%. The current waveforms were not changed and were simply scaled, and the effects on both occurred approximately 15 s after OAG was applied. In chick embryo DRGs OAG inhibited the T current by 30% and the L current by 50%. We conclude that PKC modulates Ca2+ currents by acting on both L and T Ca2+ channels.


A spectrum of severe familial liver disorders associate with telomerase mutations.
Telomerase is an enzyme specialized in maintaining telomere lengths in highly proliferative cells. Loss-of-function mutations cause critical telomere shortening and are associated with the bone marrow failure syndromes dyskeratosis congenita and aplastic anemia and with idiopathic pulmonary fibrosis. Here, we sought to determine the spectrum of clinical manifestations associated with telomerase loss-of-function mutations. Sixty-nine individuals from five unrelated families with a variety of hematologic, hepatic, and autoimmune disorders were screened for telomerase complex gene mutations; leukocyte telomere length was measured by flow fluorescence in situ hybridization in mutation carriers and some non-carriers; the effects of the identified mutations on telomerase activity were determined; and genetic and clinical data were correlated. In six generations of a large family, a loss-of-function mutation in the telomerase enzyme gene TERT associated with severe telomere shortening and a range of hematologic manifestations, from macrocytosis to acute myeloid leukemia, with severe liver diseases marked by fibrosis and inflammation, and one case of idiopathic pulmonary fibrosis but not with autoimmune disorders. Additionally, we identified four unrelated families in which loss-of-function TERC or TERT gene mutations tracked with marrow failure, pulmonary fibrosis, and a spectrum of liver disorders. These results indicate that heterozygous telomerase loss-of-function mutations associate with but are not determinant of a large spectrum of hematologic and liver abnormalities, with the latter sometimes occurring in the absence of marrow failure. Our findings, along with the link between pulmonary fibrosis and telomerase mutations, also suggest a common pathogenic mechanism for fibrotic diseases in which defective telomere repair plays important role.


Dietary Intake in the Lifelines Cohort Study: Baseline Results from the Flower Food Frequency Questionnaire among 59,982 Participants.
The role of nutrition in health and disease is well established. However, more research on this topic is needed to fill gaps in our current knowledge. The Lifelines cohort study, a large Dutch prospective cohort study, was established as a resource for international researchers, aiming to obtain insight into the aetiology of healthy ageing. The study started with 167,729 participants, covering three generations, aiming to follow them for thirty years. This article describes the habitual dietary intake, assessed using the Flower Food Frequency Questionnaire (FFQ), among Lifelines cohort study participants at baseline, stratified by sex and different categories of age, socioeconomic status (SES) and body mass index (BMI). A total of 59,982 adults (23,703 men and 36,279 women), who completed the Flower FFQ and reported plausible habitual dietary intake, were included in the analyses. Median daily energy intake was higher in men (2368 kcal) than in women (1848 kcal), as well as macronutrient intake. Energy and macronutrient intake decreased with increasing age and BMI categories; no differences were observed between SES categories. Intake of most micronutrients was higher in men than in women. Differences were observed between age categories, but not between SES and BMI categories. Food groups were consumed in different amounts by men and women; differences between age, SES and BMI categories were observed as well. The Lifelines cohort study provides extensive dietary intake data, which are generalisable to the general Dutch population. As such, highly valuable dietary intake data are available to study associations between dietary intake and the development of chronic diseases and healthy aging.


Cholangiocarcinoma--controversies and challenges.
Cholangiocarcinomas are a diverse group of tumors that are presumed to originate from the biliary tract epithelium either within the liver or the biliary tract. These cancers are often difficult to diagnose, their pathogenesis is poorly understood, and their dismal prognosis has resulted in a nihilistic approach to their management. The two major clinical phenotypes are intrahepatic, mass-forming tumors and large ductal tumors. Among the ductal cancers, lesions at the liver hilum are most prevalent. The risk factors, clinical presentation, natural history and management of these two types of cholangiocarcinoma are distinct. Efforts to improve outcomes for patients with these diseases are affected by several challenges to effective management. For example, designations based on anatomical characteristics have been inconsistently applied, which has confounded analysis of epidemiological trends and assessment of risk factors. The evaluation of therapeutic options, particularly systemic therapies, has been limited by a lack of appreciation of the different phenotypes. Controversies exist regarding the appropriate workup and choice of management approach. However, new and emerging tools for improved diagnosis, expanded indications for surgical approaches, an emerging role for locoregional and intrabiliary therapies and improved systemic therapies provide optimism and hope for improved outcomes in the future.


Multifunctional biomimetic nanoparticles loading baicalin for polarizing tumor-associated macrophages.
Immunosuppression and immune tolerance lead tumor cells to evade immune system surveillance and weaken drug efficacy. The presence of various immunosuppressive cells in the tumor microenvironment, especially tumor-associated macrophages (TAMs), has been shown to be a driving force in tumor initiation and development. Reversion of the TAM phenotype is an effective way to induce a subsequent antitumor immune response. In this study, we developed baicalin-loaded poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles containing an antigenic peptide (Hgp 10025-33, Hgp) and a toll-like receptor 9 agonist (CpG). The nanoparticles were further coated with a galactose-inserted erythrocyte membrane, which actively targeted the TAMs. The TAM polarization and tumor treatment effectiveness of the nanoparticles were evaluated. The biomimetic nanoparticles showed enhanced cell uptake in vitro and targeted effects in vivo. In addition, compared with baicalin-loaded PLGA-NPs (B@NPs), the biomimetic nanoparticles, such as Hgp/B@NPs-CpG and NPs@RBC-Gala, significantly polarized the TAMs such that they changed from the M2 type to the M1 type both in vitro and in vivo. Subsequently, the infiltration of CD4+ T and CD8+ T cells into tumor sites after being induced by the biomimetic nanoparticles was greatly increased, which suggested a significant enhancement of the immune activation effect and T cell response. In addition, the activation of the T cells and induction of the CTL responses effectively suppressed melanoma tumor growth in vivo. In conclusion, the biomimetic nanoparticles effectively reversed the TAM phenotype from M2 to M1, which further improved the tumor immune microenvironment and promoted tumor immunotherapy. These results suggested that the TAM-targeted biomimetic drug delivery system had the potential to reverse the phenotypes of TAMs contributing to reverse the immunosuppressive tumor microenvironment and promote tumor treatment.


Cardioprotection against ischemia/reperfusion injury and chromogranin A-derived peptides.
Chromogranin A (CgA) is produced by cells of the sympathoadrenal system and by human ventricular myocardium. In the clinical setting CgA has been mainly used as a marker of neuroendocrine tumors, but in the last decade a plenty of data have been published on the role of CgA and its derived peptides, particularly catestatin and vasostatin, in the regulation of cardiovascular function and diseases, including heart failure and hypertension. CgA-derived peptides, namely catestatin and vasostatin, may exert negative inotropic and lusitropic effects on mammalian hearts. As such CgA and its derived peptides may be regarded as mediators of a complex feedback system able to modulate the exaggerated release of catecholamines. This system may be also interpreted as an attempt for compensatory cardioprotective response against myocardial injury in the pre and postischemic scenarios. In fact, while vasostatin can trigger cardioprotective effects akin ischemic preconditioning (protection is triggered before ischemia), catestatin is a potent cardioprotective agent in the early post-ischemic phase, acting like a postconditioning agent (protection is triggered at the onset of reperfusion). Admittedly, the exact mechanism of cardioprotection of this system is far from being fully understood. Interestingly, both vasostatin and catestatin have shown to be able to activate multiple cardioprotective pathways. In particular, these two CgA-derived peptides may induce nitric oxide dependent pathway, which may play a pivotal role in cardioprotection against ischemia/reperfusion injury. Here, we review the literature about the cardiac effects of catestatin and vasostatin, the mechanisms of myocardial injury and protection and the role of CgA derived peptides in cardioprotection.


Aryl hydrocarbon receptor-dependent stanniocalcin 2 induction by cinnabarinic acid provides cytoprotection against endoplasmic reticulum and oxidative stress.
The aryl hydrocarbon receptor (AhR) is a cytosolic ligand-activated transcription factor historically known for its role in xenobiotic metabolism. Although AhR activity has previously been shown to play a cytoprotective role against intrinsic apoptotic stimuli, the underlying mechanism by which AhR confers cytoprotection against apoptosis is largely unknown. Here, we demonstrate that activation of AhR by the tryptophan catabolite cinnabarinic acid (CA) directly upregulates expression of stanniocalcin 2 (Stc2) to elicit cytoprotection against apoptosis induced by endoplasmic reticulum stress and oxidative stress. Chromatin immunoprecipitation studies demonstrated that CA treatment induces direct AhR binding to a region of the Stc2 promoter containing multiple xenobiotic response elements. Using isolated primary hepatocytes from AhR wild-type (AhR floxed) and liver-specific AhR conditional knockout mice, we showed that pretreatment with CA conferred cytoprotection against hydrogen peroxide (H(2)O(2))-, thapsigargin-, and ethanol-induced apoptosis in an AhR-dependent manner. Furthermore, suppressing Stc2 expression using RNA interference confirmed that the cytoprotective properties of CA against H(2)O(2), thapsigargin, and ethanol injury were absolutely dependent on Stc2. Immunochemistry revealed the presence of Stc2 in the endoplasmic reticulum and on the cell surface, consistent with Stc2 secretion and autocrine and/or paracrine signaling. Finally, in vivo data using a mouse model of acute alcohol hepatotoxicity demonstrated that CA provided cytoprotection against ethanol-induced apoptosis, hepatic microvesicular steatosis, and liver injury. Collectively, our data uncovered a novel mechanism for AhR-mediated cytoprotection in the liver that is dependent on CA-induced Stc2 activity.


Organic chemistry. A data-intensive approach to mechanistic elucidation applied to chiral anion catalysis.
Knowledge of chemical reaction mechanisms can facilitate catalyst optimization, but extracting that knowledge from a complex system is often challenging. Here, we present a data-intensive method for deriving and then predictively applying a mechanistic model of an enantioselective organic reaction. As a validating case study, we selected an intramolecular dehydrogenative C-N coupling reaction, catalyzed by chiral phosphoric acid derivatives, in which catalyst-substrate association involves weak, noncovalent interactions. Little was previously understood regarding the structural origin of enantioselectivity in this system. Catalyst and substrate substituent effects were probed by means of systematic physical organic trend analysis. Plausible interactions between the substrate and catalyst that govern enantioselectivity were identified and supported experimentally, indicating that such an approach can afford an efficient means of leveraging mechanistic insight so as to optimize catalyst design.


[Psychologico-medical studies in intensive care].
Depending to a certain extent on the nature and phase of the disease, or the duration of intensive care, by far the greater percentage of patients report positively on their experience in the ICU. In contrast, the members of the care-providing team expect negative evaluations on the part of their patients, adopt a very critical attitude towards their own work, and are involved in numerous interpersonal conflicts. The studies reveal numerous stresses, with both internal and external causes, for patients and staff, impairment, or even disruption, of meaningful communication within the team, as also between members of the team and patients. A study of the various individual realities, challenges and possibilities for coping on the part of the patients, nurses and doctors confronted with disease, suffering, dying and death, or recovery, points to differences in the regulation of (interpersonal) closeness and distance as an important source of conflict.


Early functional muscle regeneration after myotoxic injury in mice is unaffected by nNOS absence.
Nitric oxide (NO) is an important signaling molecule produced in skeletal muscle primarily via the neuronal subtype of NO synthase (NOS1, or nNOS). While many studies have reported NO production to be important in muscle regeneration, none have examined the contribution of nNOS-derived NO to functional muscle regeneration (i.e., restoration of the muscle's ability to produce force) after acute myotoxic injury. In the present study, we tested the hypothesis that genetic deletion of nNOS would impair functional muscle regeneration after myotoxic injury in nNOS(-/-) mice. We found that nNOS(-/-) mice had lower body mass, lower muscle mass, and smaller myofiber cross-sectional area and that their tibialis anterior (TA) muscles produced lower absolute tetanic forces than those of wild-type littermate controls but that normalized or specific force was identical between the strains. In addition, muscles from nNOS(-/-) mice were more resistant to fatigue than those of wild-type littermates (P < 0.05). To determine whether deletion of nNOS affected muscle regeneration, TA muscles from nNOS(-/-) mice and wild-type littermates were injected with the myotoxin notexin to cause complete fiber degeneration, and muscle structure and function were assessed at 7 and 10 days postinjury. Myofiber cross-sectional area was lower in regenerating nNOS(-/-) mice than wild-type controls at 7 and 10 days postinjury; however, contrary to our original hypothesis, no difference in force-producing capacity of the TA muscle was evident between the two groups at either time point. Our findings reveal that nNOS is not essential for functional muscle regeneration after acute myotoxic damage.


Expression of the ether-a-go-go (ERG) potassium channel in smooth muscle of the equine gastrointestinal tract and influence on activity of jejunal smooth muscle.
To determine whether ether-a-go-go (ERG) potassium channels are expressed in equine gastrointestinal smooth muscle, whether ERG channel antagonists affect jejunal muscle contraction in vitro, and whether plasma cisapride concentrations in horses administered treatment for postoperative ileus (POI) are consistent with ERG channels as drug targets. Samples of intestinal smooth muscle obtained from 8 horses free of gastrointestinal tract disease and plasma samples obtained from 3 horses administered cisapride for treatment of POI. Membranes were prepared from the seromuscular layer of the duodenum, jejunum, ileum, cecum, large colon, and small colon. Immunoblotting was used to identify the ERG channel protein. Isolated jejunal muscle strips were used for isometric stress response to ERG channel blockers that included E-4031, MK-499, clofilium, and cisapride. Plasma concentrations of cisapride were determined in 3 horses administered cisapride for treatment of POI after small intestinal surgery. Immunoblotting identified ERG protein in all analyzed segments of the intestinal tract in all horses. The selective ERG antagonist E-4031 caused a concentration-dependent increase in jejunal contraction. Clofilium, MK-499, and cisapride also increased jejunal contraction at concentrations consistent with ERG channel block; effects of E-4031 and cisapride were not additive. Peak plasma cisapride concentrations in treated horses were consistent with ERG block as a mechanism of drug action. The ERG potassium channels modulate motility of intestinal muscles in horses and may be a target for drugs. This finding may influence development of new prokinetic agents and impact treatment of horses with POI.


The cytopathology of malignant peripheral nerve sheath tumor: a report of 55 fine-needle aspiration cases.
Among sarcomas, a diagnosis of malignant peripheral nerve sheath tumor (MPNST) is often one of exclusion due to the absence of unequivocally characteristic histopathology, a conclusive immunohistochemical profile, or even a unique chromosomal anomaly. Because of this, the fine-needle aspiration (FNA) cytopathology of MPNST is extremely challenging. In the current study, the authors review their FNA experience with this neoplasm. The authors searched their combined departmental cytology files for all lesions signed out as MPNST or suspicious for MPNST, as well as their own surgical pathology files for any cases of MPNST that had corresponding cytology. FNA was performed using standard techniques. A total of 55 cases of MPNST, all with tissue confirmation, and 1 misdiagnosed example of melanoma were retrieved from 52 patients (M:F ratio of 1.5:1; average age, 46 years), 26 of whom had a history of neurofibromatosis type 1 (NF-1). Aspirates were from primary (27 cases), locally recurrent (14 cases), or metastatic (10 cases) MPNST; 4 primary tumor aspirates were of ex vivo specimens. Sites included the extremities (22 cases), trunk/pelvis (22 cases), head and neck (6 cases), and deep-seated masses (6 cases). FNA diagnoses were MPNST (24 patients); consistent with MPNST (5 patients); sarcoma, not otherwise specified (10 patients); atypical (3 patients); spindle cell neoplasm (6 patients); malignant neoplasm (1 patient); and nondiagnostic (3 patients). A definitive diagnosis of either MPNST or consistent with MPNST was issued in 30%, 93%, and 70%, respectively, of primary, locally recurrent, and metastatic lesions. FNA cytopathology is limited as a diagnostic instrument for the initial diagnosis of MPNST, but is exceedingly accurate and valuable in the recognition of metastatic and locally recurrent MPNST.


Screening for Chlamydia trachomatis using self-collected vaginal swabs at a public pregnancy termination clinic in France: results of a screen-and-treat policy.
To assess the prevalence of Chlamydia trachomatis (CT) infection and the risk factors for CT infection among women presenting for abortion at a clinic in France. Women seeking surgically induced abortions were systematically screened by PCR on self-collected vaginal swabs between January 1, 2010, and September 30, 2010. CT-positive women were treated with oral azithromycin (1 g) before the surgical procedure. Of the 978 women included in the study, 66 were CT positive. The prevalence was 6.7% (95% confidence intervals [CI] 5.1%-8.3%). The risk factors for CT infection were the following: age <30 years (Odds ratio [OR]: 2.0 [95% CI: 1.2-3.5]), a relationship status of single (OR: 2.2 [95% CI: 1.2-4.0]), having 0 or 1 child (OR: 5.2 [95% CI: 2.0-13.0]), not using contraception (OR: 2.4 [95% CI: 1.4-4.1]), and completing 11 weeks or more of gestation (OR: 2.1 [95% CI: 1.3-3.6]). Multiple logistic regression indicated that 4 factors--having 0 or 1 child, a single relationship status, no contraceptive use, and a gestation of 11 weeks or more--were independently associated with CT infection. The rate of postabortion infection among all patients was 0.4% (4/978). These results reveal a high prevalence (6.7%) of CT-positive patients among French women seeking induced abortions. Because it is not common practice to screen the general population for CT, screening before induced abortions seems relevant. A cost-effectiveness study is required to evaluate this screen-and-treat policy.


Isolation and lipid characterization of cholesterol-enriched fractions in cortical and nuclear human lens fibers.
Human lens membranes contain unusually high levels of cholesterol and sphingolipids, lipids known to segregate into liquid-ordered domains. The current study was conducted to pursue the determination and characterization of these domains in membranes of clear and cataractous human lenses. Cortical and nuclear regions of aged clear and cataractous lenses were obtained. After lysis with Triton X-100 at 4 degrees C and sucrose linear-density centrifugation, sedimenting and nonsedimenting fractions (when present) were collected. Phospholipids were analyzed by (31)P-nuclear magnetic resonance (NMR) and mass spectrometry. Caveolae and raft markers were tested by Western blot analysis. Only samples from clear lenses exhibited a nonsedimenting band. Phospholipid contents were comparable for sedimenting fractions of clear and cataractous membranes. Cholesterol to phospholipid molar ratios in light-density bands were nearly 7, three times greater than in sedimenting fractions. The portion of total cholesterol present in nonsedimenting fractions increased from 5.5% in the cortex to 14% in the nucleus. Two lysophospholipids comprising approximately 10% of all phospholipids in total membranes were undetectable in nonsedimenting fractions. Caveolin-1 was enriched in these fractions. Phospholipid compositional differences between lighter and heavier fractions from clear lenses were relatively minor and could not, alone, account for the substantial enrichment of cholesterol in the lighter fractions. Specific proteins, such as caveolin-1, must recruit cholesterol and induce clustering. Undetectable amounts of light-density domains in cataractous membranes suggest either disruption of these aggregates and thus the function of proteins within them, possibly relevant to lens transparency, and/or greater density of these clusters due to stronger binding of insoluble crystallins to membranes.


Interneuron-specific Ca2+ responses linked to metabotropic and ionotropic glutamate receptors in rat hippocampal slices.
Glutamate-mediated regulation of intracellular Ca2+ levels was examined in different populations of CA1 interneurons, using confocal microscopy and the Ca2+ indicator fluo 3-AM in rat hippocampal slices. Interneurons in basal [stratum oriens/alveus (OA)] and apical [strata radiatum and lacunosum-moleculare (R/LM)] dendritic layers responded heterogeneously to glutamate. In control medium, OA interneurons responded mostly with oscillatory Ca2+ responses, which consisted of a large Ca2+ transient and successive smaller elevations. R/LM interneurons responded mostly with biphasic responses, characterized by an initial large transient and a secondary prolonged elevation. Other interneurons in both R/LM and OA responded with transient elevations in Ca2+ levels. Ionotropic glutamate receptor antagonists (+/-)2-amino-5-phosphonopentanoic acid and 6-cyano-7-nitro-quinoxaline-2,3-dione reduced peak Ca2+ responses in OA and R/LM cells, and blocked biphasic responses in R/LM interneurons. The metabotropic glutamate receptor antagonist (RS)-alpha-methyl-4-carboxyphenylglycine reduced peak Ca2+ responses only in OA interneurons, and prevented oscillatory responses. In low Ca2+ medium, peak responses were reduced in R/LM but not in OA interneurons, and oscillatory responses were absent. Combination of ionotropic and metabotropic receptor antagonists blocked all glutamate-evoked Ca2+ responses. Activation of different types of glutamate receptors may thus produce heterogeneous Ca2+ signals in subpopulations of CA1 interneurons. Ionotropic receptors may generate biphasic responses in interneurons in apical dendritic layers, whereas combined activation of metabotropic and ionotropic receptors may trigger oscillatory responses in interneurons of basal dendritic layers. These heterogeneous Ca2+ responses indicate that glutamate-mediated Ca2+ processes and second messenger systems differ in subpopulations of hippocampal interneurons and suggest possible postsynaptic functional specialization of interneurons.


The role of flumazenil in outpatient transesophageal echocardiography.
Benzodiazepines used for transesophageal echocardiography (TEE) sedation may be associated with postprocedural psychomotor effects that are undesirable. We hypothesize that flumazenil can reverse cognitive and motor effects from benzodiazepine, promoting earlier return to baseline function. We prospectively evaluated the cognitive and motor function of patients who did or did not receive flumazenil following TEE. Patients' gait, level of drowsiness, and recall of items learned before and after benzodiazepine administration were evaluated before TEE, as well as immediately and 30 min after the procedure. Of 207 patients (123 men and 84 women), 93 (45%) were given flumazenil 0.2 or 0.4 mg intravenously, and 113 (55%) were not. The baseline characteristics of the patients who received flumazenil were not significantly different from those who did not receive flumazenil, with the exception of a higher mean dosage of midazolam administered to the flumazenil group. In addition, patients in the flumazenil group appeared more drowsy immediately following TEE. When adjusted for age and midazolam dosage, there were no differences, at any time, between the two groups in gait or recall of items learned prior to sedation. however, at 30 min following TEE, the flumazenil group was able to recall a larger number of new items learned immediately after the procedure (1.92/3 vs. 1.61/3, p = 0.02) than did patients in the group not receiving flumazenil. No adverse effects were encountered in any patient. Flumazenil appears safe and effective in reversing anterograde amnesic effects of benzodiazepine following TEE, but has no effects on retrograde amnesia and does not promote earlier return of motor function to baseline. It is useful in clinical situations where high dosages of benzodiazepine have been used and/or excessive drowsiness is evident following TEE. Routine use of the drug, however, is not necessary.


Proteome-based biomarkers in pancreatic cancer.
Pancreatic cancer, as a highly malignant cancer and the fourth cause of cancer-related death in world, is characterized by dismal prognosis, due to rapid disease progression, highly invasive tumour phenotype, and resistance to chemotherapy. Despite significant advances in treatment of the disease during the past decade, the survival rate is little improved. A contributory factor to the poor outcome is the lack of appropriate sensitive and specific biomarkers for early diagnosis. Furthermore, biomarkers for targeting, directing and assessing therapeutic intervention, as well as for detection of residual or recurrent cancer are also needed. Thus, the identification of adequate biomarkers in pancreatic cancer is of extreme importance. Recently, accompanying the development of proteomic technology and devices, more and more potential biomarkers have appeared and are being reported. In this review, we provide an overview of the role of proteome-based biomarkers in pancreatic cancer, including tissue, serum, juice, urine and cell lines. We also discuss the possible mechanism and prospects in the future. That information hopefully might be helpful for further research in the field.


Long non-coding RNA placenta‑specific protein 2 regulates the chemosensitivity of cancer cells to cisplatin in hepatocellular carcinoma (HCC) by sponging microRNA-96 to upregulate X-linked inhibitor of apoptosis protein.
This study was conducted to investigate the roles of lncRNA PLAC2 and XiaP in hepatocellular carcinoma (HCC). HCC and paired non-tumor tissues were collected from 62 HCC patients who received cisplatin-based treatment. At 0, 2, and 4 months of post-cisplatin-based therapy, blood samples (5 ml) were collected from all patients and prepared plasma samples. LncRNA PLAC2 expression in tissue and plasma samples was determined by RT-qPCR. The interactions between lncRNA PLAC2 and XiaP in HCC cell lines were assessed by overexpression experiments. Cell viability and apoptosis under cisplatin treatment were analyzed by MTT assay and cell apoptosis assay, respectively. The direct interaction between lncRNA PLAC2 and miR-96, which can target XiaP, was analyzed by performing RNA-RNA pulldown assay. It was observed that lncRNA PLAC2 was upregulated in HCC tissues than in non-tumor tissues. LncRNA PLAC2 expression in HCC tissues was not affected by HBV and HCV but upregulated after cisplatin-based treatment. Similarly, cisplatin treatment of HCC cells increased PLAC2 expression. LncRNA PLAC2 and XiaP overexpression increased viability and decreased apoptosis of cisplatin-treated HCC cells, while lncRNA PLAC2 knockdown decreased viability and increased apoptosis of cisplatin-treated HCC cells. Western blot analysis showed that lncRNA PLAC2 increased XiaP protein accumulation, while lncRNA PLAC2 siRNA silencing decreased XiaP expression in HCC cells. LncRNA PLAC2 and miR-96 directly interacted with each other, while they failed to regulate the expression of each other. In conclusion, lncRNA PLAC2 negatively regulates the chemosensitivity of HCC cells to cisplatin, possibly by sponging miR-96 to upregulate miR-96.


Lymph node involvement by mycosis fungoides and Sézary syndrome mimicking angioimmunoblastic T-cell lymphoma.
Clinical management of cutaneous T-cell lymphoma (CTCL) and angioimmunoblastic T-cell lymphoma (AITL) differs markedly. Diagnostic distinction is critical. Herein, we describe a series of 4 patients with clinically, molecularly, and histopathologically annotated mycosis fungoides or Sézary syndrome whose nodal disease mimicked AITL. The patients otherwise exhibited classic clinical manifestations of mycosis fungoides/Sézary syndrome preceding the onset of lymphadenopathy by 1 to 5 years. Skin biopsies revealed epidermotropic infiltrates characteristic of CTCL. Lymph node biopsies revealed dense CD4+ T-cell infiltrates that coexpressed follicular helper T-cell markers and were accompanied by proliferations of high endothelial venules and arborizing CD21+ follicular dendritic cell networks. Two patients had T-cell receptor gene rearrangement studies performed on their skin, lymph node, and peripheral blood demonstrating identical polymerase chain reaction clones in all 3 tissues. A small secondary clonal B-cell population was present in 1 patient that mimicked the B-cell proliferations known to accompany AITL and persisted on successive nodal biopsies over several years. This latter phenomenon has not previously been described in CTCL. The potential for patients to be misdiagnosed with AITL for lack of consideration of advanced-stage CTCL with nodal involvement underscores the necessity of information sharing among the various pathologists and clinicians involved in the care of each patient.


Linkage analysis of five Chinese families with arrhythmogenic right ventricular cardiomyopathy using microsatellite genetic markers.
To explore the linkage relationship between specific genetic markers and arrhythmogenic right ventricular cardiomyopathy (ARVC) in Chinese pedigrees. The microsatellite genetic markers D2S152, D14S252, and D10S1664 were studied for their linkages to ARVC in five Chinese ARVC pedigrees and a normal population of 121 Chinese individuals. Genomic DNA of the pedigrees and normal population was amplified using PCR techniques. Denaturing polyacrylamide sequencing gel (4%) electrophoresis was used to detect microsatellite repeat polymorphisms. Gels were silver-stained. A classical linkage analysis program was used assuming models of autosomal dominance and recession. The logarithm of the odds (LOD) scores of D2S152 with ARVC in LW, WD, DS, LC and TY pedigrees were 2.174, -0.589, -infinity, - (indicating that linkage is not supported in this mode), and -infinity respectively in autosomal dominant model (recombination fraction = 0.000 respectively)and were -infinity, -infinity, -infinity, -infinity, and 0.182 respectively in the autosomal recessive model. The LOD scores of D14S252 with ARVC in LW, WD, DS, LC and TY pedigrees were -, -, -infinity, -, and 0 respectively in autosomal dominant model, and were -infinity, -0.812, -infinity, -infinity, and 0.087 respectively in autosomal recessive model. The LOD scores of D2S152 with ARVC in LW, WD, DS, LC and TY pedigrees were -, -0.539, -, and 0.602 respectively in autosomal dominant model and were -, -infinity, -infinity, -infinity, and - infinity respectively in autosomal recessive model. The LOD score for D2S152 in the LW pedigree was 2.174, indicating that the chance of linkage is about 150:1. This suggests that there is a possible ARVC-related gene near this marker. There were no clear linkage relationships between ARVC and D10S1664 and D14S252 in this family, and no linkages between ARVC and any of the three genetic markers in the other four families. These results also suggest that there is genetic heterogeneity in LW and in the other pedigrees.


Effect of breast feeding on ocular morbidity.
To evaluate the effect of human milk on ocular morbidity in infants who did not breast feed during the first 6 months of life. This retrospective randomized study included 154 subjects who were first or second grade elementary school students, 66-84 month of age. These children were randomly selected from 2080 students during health screenings. All of them were born at term and were normal birth weight. The study was composed of 3 age- and sex-matched groups. Group 1 included 52 subjects who had never been breast fed. Group 2 included 42 subjects who were breast fed and also received additional food (infant formula). Group 3 included 60 subjects who were only breast fed until 6 months old (no formula) except for the first month of life. All subjects underwent a complete ophthalmic examination and any morbidity was recorded. Frequencies of ocular morbidity were compared among the groups by using the chi-squared test. We found significant refractive errors in 12 (23%) subjects in Group 1 (no breast feeding). There was no significant refractive error in Group 2 (breast feeding and formula) and Group 3 (breast feeding only). The difference among the groups was statistically significant (p=0.014, chi-squared test). Allergic conjunctivitis was found in 5 subjects in Group 1, 3 in Group 2, and 2 in Group 3. There was no significant difference among the groups (p=0.395). Refractive errors were more frequent in Group 1 (no breast feeding) than in Group 2 (breast feeding and formula) or Group 3 (breast feeding only). There is a need to confirm this finding by performing studies with larger sample sizes.


Collaborative Public Health Investigation of Clenbuterol-Adulterated Heroin Outbreak-Richmond, Virginia, March-April 2015.
In March 2015, the Virginia Department of Health (VDH) was alerted by the Virginia Poison Center of a 6-patient cluster treated for severe clinical presentations after using heroin. Patients' symptoms were atypical for heroin use, and concern existed that patients were exposed to heroin that had been adulterated with or replaced by another substance. To understand the extent and characterization of the outbreak and implement response measures to prevent further cases. The purpose of this report is to highlight the collaborative nature of a public health investigation among a diverse group of stakeholders. Active surveillance and retrospective case finding. Richmond metro area community and hospitals. Regional poison centers, the Division of Consolidated Laboratory Services, the Department of Behavioral Health and Developmental Services, community partners, local law enforcement, and multiple VDH divisions. Outbreak investigation, communication to public health professionals, clinicians, and the community, and liaising with the local law enforcement. Outbreak control. Laboratory confirmation of clenbuterol in clinical specimens implicated it as the heroin adulterant. Thirteen patients met clinical and epidemiologic criteria for exposure to clenbuterol-adulterated heroin. All patients were associated with a localized area within Richmond, and patient interviews elucidated heroin supplier information. VDH collaborated with local law enforcement agents who investigated and arrested the supplier, leading to cessation of the outbreak. This outbreak highlights the value of policies and practices that support an integrated outbreak response among public health practitioners, poison center staff, laboratorians, clinicians, law enforcement agents, community groups, and other agencies. Collaboration enabled implementation of effective control measures-including those outside the purview of the health department-and should be standard practice in future outbreaks involving illicit substances.


Kinetics of Ca2+-activated K+ channels from rabbit muscle incorporated into planar bilayers. Evidence for a Ca2+ and Ba2+ blockade.
The interaction of Ca2+ and Ba2+ with a Ca2+-activated K+ channel from rabbit skeletal muscle membranes is studied in planar lipid bilayers. At [Ca2+] greater than or equal to 100 microM in the cis side (the side to which the vesicles are added) and at positive voltages, the channel kinetics consisted of bursts of activity interrupted by long periods of quiescence. We found that the reciprocal of the mean burst time increases linearly with [Ca2+], whereas the mean time for the quiescent (closed) periods is independent of [Ca2+]. The number of quiescent periods is reduced by increasing [K+]. Micromolar amounts of cis Ba2+ do not activate the channel, but induce similar "slow" closings. Also, in this case, the mean burst time is inversely proportional to the [Ba2+] and the mean closed time is independent of [Ba2+]. Raising [K+] either symmetrically or only in the trans side relieved the Ba2+ effect. trans Ba2+ also induces changes in the slow kinetics, but in millimolar amounts. These results suggest that the quiescent periods correspond to a channel blocked by a Ba ion. The voltage dependence of the cis blockade indicates that the Ba2+ binding site is past the middle of the membrane field. The similarities in the slow kinetics induced by Ca2+ and Ba2+ suggest that Ca2+ blocks the channel by binding to the same site. However, binding of Ca2+ to the site is 10(5)-fold weaker.


Copper sensitivity in dorsal hippocampus slices.
The action of copper on the pyramidal neurons in CA1 of the hippocampus is little understood. Our main aim was to study the possible interaction of copper on the synaptic network in CA1 pyramidal neurons. We used Wistar rats hippocampus slices in a recording chamber. The population response ("population of spikes") collected by an extracellular micropipette under baseline conditions served as control. Copper, GABA, bicuculline and picrotoxin were delivered in different experimental conditions to the slice. One, 10 and 100 microM of copper concentration decreased significantly the amplitude and duration of the population spikes in relation to the control response. This effect did not show concentration dependency. Copper in bicuculline medium decreased significantly the duration response in relation to the control response and in relation to copper effect in a free bicuculline medium. This phenomenon emphasizes the copper action on the GABA (B) and (C) receptors. Copper in a picrotoxin medium increased significantly the excitability of the response. This new effect suggests that copper acts on non-GABA receptors, an effect that could be detected when the GABA receptors were inactivated. As a result of these findings it appears that, under our experimental conditions, copper generated transient sensitivity changes in pyramidal neurons of CA1 dorsal hippocampus.


[The evaluation of noncompliance in an anti-influenza vaccination program].
The article presents an evaluation of an anti-influenza vaccination programme in a population at risk, with a special study of the general characteristics of the group of non-complying patients, and the possible relation of these characteristics with their conduct. Moreover, a study was made of the possible motives for not complying with the vaccination programme by means of a survey carried out by post and telephone in a representative sample of 108 individuals. A 34.77% of the registered population did not receive the anti-influenza vaccine, corresponding largely with the youngest age group (15 to 44 years) and there were no significant differences determined by sex or risk factor or doctor. A positive response was received from 75% of those surveyed, and the basic conclusions drawn were a low proportion of information error attributable to the programme (2.94%), and patient refusal as the principal cause of non-compliance (54.41%), followed by "other reasons" (hospitalisation, absence from home, etc.) (32.35%). The strategies that may be followed in the face of this problem require definition of the group of non-compliers, further understanding of the motives for their conduct, and educational measures to change their attitude. 95.9%). Incidence of hypertension and hyperlipemia was 56.2% and 47.3%. Late vascular events varied between 2.6% (nephropathy) and 19.5% (retinopathy). A total of 101 patients (44.6%) were unaware of the existence of diabetic health education programmes. Moreover, 68 (30.3%) declared that they had never received any previous information about their diabetes. Practical skills were evaluated on an individual basis.


Palladium-catalyzed cross-coupling of H-phosphinate esters with chloroarenes.
The palladium-catalyzed cross-coupling reaction between H-phosphinate esters and chloroarenes or chloroheteroarenes is described. This reaction is the first general metal-catalyzed phosphorus-carbon bond-forming reaction between a phosphorus nucleophile and chloroarenes.


Site of airway obstruction in asthma as determined by measuring maximal expiratory flow breathing air and a helium-oxygen mixture.
Because maximum expiratory flow-volume rates in normal subjects are dependent on gas density, the resistance between alveoli and the point at which dynamic compression begins (R(us)) is mostly due to convective acceleration and turbulence. We measured maximum expiratory flow-volume (MEFV) curves in asthmatics and chronic bronchitics breathing air and He-O(2). In the latter and in some asthmatics, MEFV curves did not change, indicating that R(us) is mostly due to laminar flow. Therefore, the point at which dynamic compression begins must be further upstream than in normal subjects and the site of obstruction must be in small airways. In other asthmatics, flow increased normally indicating obstruction in larger airways. The response to He-O(2) did not correlate with initial values of pulmonary resistance, the initial MEFV curves or the response to bronchodilators. We conclude that the site of airway obstruction varies among asthmatics and that the site of obstruction is not detectable by measurement of the usual parameters of lung mechanics.


Immunosuppressive agents for treating IgA nephropathy.
IgA nephropathy (IgAN) is a world-wide disease and the cause of end-stage renal failure (ESRF) in 15 to 20% of patients within 10 years and in 30 to 40% of individuals within 20 years from the apparent onset of disease. No specific treatment has yet been established but many approaches have been investigated. To assess the benefits and harms of immunosuppressive treatment for IgAN. We searched The Cochrane Renal Group's specialized register (May 2003), Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, Issue 3, 2002) MEDLINE (1966 - September 2002), EMBASE (1988 - September 2002) and handsearched reference lists of retrieved articles and conference proceedings. Randomized controlled trials (RCTs) and quasi-RCTs comparing treatment of IgAN with immunosuppressive agents against placebo, no treatment, other immunosuppressive or non-immunosuppressive agents. Two reviewers independently assessed trial quality and extracted data. Statistical analyses were performed using the random effects model and the results expressed as relative risk (RR) for dichotomous outcomes and weighted mean difference (WMD) for continuous outcomes, with 95% confidence intervals (CI). Thirteen eligible RCTs involving 623 patients were identified. All identified RCTs had a placebo, no treatment or warfarin/dipyridamole control group. Seven trials used steroids, three used alkylating agents/cyclosporin and three used combinations of steroids and alkylating agents/cyclosporin. No trial directly compared steroids versus alkylating agents/cyclosporin. Quality was sub-optimal. Steroids were associated with a lower risk of progression to ESRF (RR 0.44, 95% CI 0.25 to 0.80) and lower urinary protein excretion (WMD -0.49 g/24h, 95% CI -0.72 to -0.12). Urinary protein excretion was lower for patients treated with alkylating agents/cyclosporin compared to placebo/no treatment (WMD -0.94 g/24h, 95% CI -1.43 to -0.46). There was no significant reduction of urinary protein excretion with combination treatment of steroids and alkylating agents compared with placebo/no treatment. The optimal management of IgAN remains uncertain. The RCTs identified were small, of sub-optimal methodological quality and tended to only report favorable and surrogate outcomes without a thorough reporting of treatment harms. All outcomes favor the use of immunosuppressive interventions, with steroids appearing to be the most promising. Further study, in the form of RCTs, is necessary to ascertain which patients would benefit from these interventions, whether they are the ones with early signs of renal dysfunction or those with more advanced renal impairment.


Glycaemia risk index uncovers distinct glycaemic variability patterns associated with remission status in type 1 diabetes.
The aim of this work was to define a unique remission status using glycaemia risk index (GRI) and other continuous glucose monitoring (CGM) metrics in individuals with type 1 diabetes for improved phenotyping. A group of 140 individuals with type 1 diabetes were recruited for a cross-sectional study. The participants were categorised into four groups based on their remission status, which was defined as insulin-dose-adjusted A1c (IDAA1c) <9 or C-peptide ≥300 pmol/l: new-onset (n=24); mid-remission (n=44); post-remission (n=44); and non-remission (individuals who did not experience remission, n=28). Participants in the remission phase were referred to as 'remitters', while those who were not in the remission phase were referred to as 'non-remitters', the latter group including new-onset, post-remission and non-remission participants. Clinical variables such as HbA1c, C-peptide and insulin daily dose, as well as IDAA1C and CGM data, were collected. The patterns of CGM metrics were analysed for each group using generalised estimating equations to investigate the glycaemic variability patterns associated with remission status. Then, unsupervised hierarchical clustering was used to place the participants into subgroups based on GRI and other CGM core metrics. The glycaemic variability patterns associated with remission status were found to be distinct based on the circadian CGM metrics. Remitters showed improved control of blood glucose levels over 14 days within the range of 3.9-10 mmol/l, and lower GRI compared with non-remitters (p<0.001). Moreover, GRI strongly correlated with IDAA1C (r=0.62; p<0.001) and was sufficient to distinguish remitters from non-remitters. Further, four subgroups demonstrating distinct patterns of glycaemic variability associated with different remission status were identified by clustering on CGM metrics: remitters with low risk of dysglycaemia; non-remitters with high risk of hypoglycaemia; non-remitters with high risk of hyperglycaemia; and non-remitters with moderate risk of dysglycaemia. GRI, an integrative index, together with other traditional CGM metrics, helps to identify different glycaemic variability patterns; this might provide specifically tailored monitoring and management strategies for individuals in the various subclusters.


Overexpression of tau in a nonneuronal cell induces long cellular processes.
The ways in which the various microtubule-associated proteins (MAPs) contribute to cellular function are unknown beyond the ability of these proteins to modify microtubule dynamics. One member of the MAP family, tau protein, is restricted in its distribution to the axonal compartment of neurons, and has therefore prompted studies that attempt to relate tau function to the generation or maintenance of this structure. Sf9 cells from a moth ovary, when infected with a baculovirus containing a tau cDNA insert, elaborate very long processes. This single gene product expressed in a foreign host cell grossly alters the normal rounded morphology of these cells. The slender, relatively nonbranched appearance of these processes as well as their uniform caliber resembles the light-microscopic appearance of axons observed in several neuronal culture systems. Immunolabeling of the tau-expressing Sf9 cells demonstrated tau reactivity in the induced processes, and EM that microtubule bundles were present in the processes. Microtubule stabilization alone was insufficient to generate processes, since taxol treatment did not alter the overall cell shape, despite the induction of microtubule bundling within the cell body.


Tacrolimus Protects against Age-Associated Microstructural Changes in the Beagle Brain.
The overexpression of calcineurin leads to astrocyte hyperactivation, neuronal death, and inflammation, which are characteristics often associated with pathologic aging and Alzheimer's disease. In this study, we tested the hypothesis that tacrolimus, a calcineurin inhibitor, prevents age-associated microstructural atrophy, which we measured using higher-order diffusion MRI, in the middle-aged beagle brain (n = 30, male and female). We find that tacrolimus reduces hippocampal (p = 0.001) and parahippocampal (p = 0.002) neurite density index, as well as protects against an age-associated increase in the parahippocampal (p = 0.007) orientation dispersion index. Tacrolimus also protects against an age-related decrease in fractional anisotropy in the prefrontal cortex (p < 0.0001). We also show that these microstructural alterations precede cognitive decline and gross atrophy. These results support the idea that calcineurin inhibitors may have the potential to prevent aging-related pathology if administered at middle age.SIGNIFICANCE STATEMENT Hyperactive calcineurin signaling causes neuroinflammation and other neurobiological changes often associated with pathologic aging and Alzheimer's disease (AD). Controlling the expression of calcineurin before gross cognitive deficits are observable might serve as a promising avenue for preventing AD pathology. In this study, we show that the administration of the calcineurin inhibitor, tacrolimus, over 1 year prevents age- and AD-associated microstructural changes in the hippocampus, parahippocampal cortex, and prefrontal cortex of the middle-aged beagle brain, with no noticeable adverse effects. Tacrolimus is already approved by the Food and Drug Administration for use in humans to prevent solid organ transplant rejection, and our results bolster the promise of this drug to prevent AD and aging-related pathology.


Epigenome-wide association study of maternal hemoglobin A1c in pregnancy and cord blood DNA methylation.
Background: Gestational hyperglycemia is associated with adverse perinatal outcomes and long-term offspring metabolic programming, likely through dysregulation of DNA methylation (DNAm). Materials & methods: We tested associations between maternal HbA1c and cord blood DNAm among 412 mother-child pairs in the genetics of glucose regulation in gestation and growth (Gen3G) and implemented Mendelian randomization to infer causality. We sought replication in an independent sample from Healthy Start. Results: Higher second trimester HbA1c levels were associated with lower DNAm at cg21645848 (p = 3.9 × 10-11) near URGCP. Mendelian randomization and replication analyses showed same direction of effect between HbA1c and DNAm at cg21645848, but did not reach statistical significance. Conclusion: We found that higher maternal glycemia reflected by HbA1c is associated with cord blood DNAm at URGCP, a gene related with inflammatory pathways.


[Excitation and inhibition in monosynaptic reflex arcs of white rats in conditions of spinal hyperreflexia].
The mechanisms of excitation and inhibition in monosynaptic reflex arcs of white rats in conditions of spinal superreflexion (a state, in which the amplitude of reflex monosynaptic discharges of a ventral root (MD VR) is close to a threshold of excitation of motoneurons axon) were investigated. The character of MD VR during recording of their reflex discharges from cut and uncut VR was studied. Previously established pattern MR VR generation as extremely high voltage and presence of its second component are observed in these conditions. In a state of superreflexion intersegmental interactions getting weaker and are particularly characterized by inhibitive influences from segment L4 on motoneurons of segment L5. It is shown that in animals with spinal superreflexion expression of the antidromic inhibition is weaker compare to intact ones. It was defined that reciprocal interrelations of flexor and extensor motoneurons are maintained and even strengthen.


Strengthening public health education in population and reproductive health through an innovative academic partnership in Africa: the Gates partners experience.
Poor reproductive health constitutes one of the leading public health problems in the world, particularly in sub-Saharan Africa (SSA). We report here an academic partnership that commenced in 2003 between a US institution and six universities in SSA. The partnership addresses the human resources development challenge in Africa by strengthening public health education and research capacity to improve population and reproductive health (PRH) outcomes in low-resource settings. The partnership's core activities focused on increasing access to quality education, strengthening health research capacity and translating scholarship and science into policy and practices. Partnership programmes focused on the educational dimension of the human resources equation provide students with improved learning facilities and enhanced work environments and also provide faculty with opportunities for professional development and an enhanced capacity for curriculum delivery. By 2007, 48 faculty members from the six universities in SSA attended PRH courses at Johns Hopkins University, 93 PRH courses were offered across the six universities, 625 of their master's students elected PRH concentrations and 158 had graduated. With the graduation of these and future student cohorts, the universities in SSA will systematically be expanding the number of public health practitioners and strengthening programme effectiveness to resolve reproductive health needs. Some challenges facing the partnership are described in this article.


Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye.
To identify the proteomic alterations associated with carcinogenesis of hepatocellular carcinoma (HCC), we compared the protein expression profiles of nine HCC cell lines with those of primary cultured hepatocytes established from five individuals. A differential proteomic study was performed by two-dimensional difference gel electrophoresis, in which protein samples are labeled with different fluorescent dyes and separated according to their isoelectric point and molecular weight. To label the protein samples, we used a newly developed and highly sensitive fluorescent dye, which reacts with all reduced cysteine residues of proteins. Principal component analysis based on the intensity of 1238 protein spots indicated that the HCC cells and the normal hepatocytes had distinct proteomic profiles. The Wilcoxon test was used to determine the protein spots whose intensity was differentially regulated in the HCC cells compared with the normal hepatocytes, and mass spectrometric analysis was used to identify the proteins corresponding to the spots. The proteins identified are involved in cell cycle regulation, binding to a tumor-suppressor gene product, fatty acid binding, and regulation of translation. Western blotting with specific antibodies revealed the overexpression of PCNA, EB1 and E-FABP in HCC tissues compared with noncancerous tissues. Aberrant regulation of EB1 and E-FABP has not previously been implicated in the development of HCC.


The clinical relevance of the flow properties of blood cells.
In the recent years hemorheology has gained increasing interest in medicine and contributed to new understanding of the role of blood fluidity with regard to microcirculation especially in chronic occlusive vascular disease. Various hemorheologic factors have been uncovered as being of significance in physiologic and pathologic flow conditions. Primarily the deformability of red cells was in the center of interest and various devices mostly based on filtration systems, were introduced for the measuring of red cell's ability to deform. In the last few years the role of white cells came into play especially in conditions of low perfusion pressure in ischaemic areas. Since it is plausible that in capillaries white cells can hold up a very much larger number of red cells it is most likely that the rheological properties of white cells play a governing role in areas of ischaemia and infarction in respect of blood fluidity, flow and prognostic aspects. A new filtration device is now available which can distinguish between the resistance to flow of the rheologically predominant cell type of a suspension (initial filtration rate) and the number of rigid cells blocking the filter pores (clogging rate). Intensive studies will be necessary to clarify the rheological behaviour of the different types of white cells.


Low-volume ultrasound-guided nerve block provides inferior postoperative analgesia compared to a higher-volume landmark technique.
Ultrasound guidance reduces the required local anesthetic volume for successful peripheral nerve blockade, but it is unclear whether this impacts postoperative analgesia. This prospective, randomized, observer-blinded study tested the hypothesis that a low-volume ultrasound-guided ankle block would provide similar analgesia after foot surgery compared with a conventional-volume surface landmark technique. A total of 72 patients presenting for elective foot surgery under general anesthesia were randomized to receive a low-volume ultrasound-guided ankle block (n = 37; ropivacaine 0.5% adjacent the anterior/posterior tibial arteries and short saphenous vein; subcutaneous infiltration around the saphenous and superficial peroneal nerves) or conventional-volume surface landmark guided technique (n = 35; 30 mL of ropivacaine 0.5%). Patients received regular postoperative acetaminophen, diclofenac, and rescue tramadol. Assessment was in the recovery room and at 24 hours for pain and tramadol consumption. Mean (SD) total local anesthetic volume for the low-volume ultrasound group was 16 (2.1) mL. Block success in the recovery room was similar between groups (low-volume ultrasound 89% versus conventional-volume landmark 80%, P = 0.34; however, during the first 24 hours, numerically rated (0-10) "average pain" (median [10-90th percentiles] = 1 [0-4] versus 0 [0-2], P = 0.01), worst pain at rest (1 [0-6] versus 0 [0-2], P = 0.03), and the proportion of patients requiring rescue tramadol (% [95% confidence interval]: 50 [34-46] versus 20 [10-36], P = 0.01) were higher in the low-volume ultrasound group. Numerically rated numbness, weakness, satisfaction, and procedural time were similar between groups. Low-volume ultrasound-guided ankle block is associated with a high block success rate after foot surgery; however, compared with a conventional volume (surface landmark) technique, the reduced local anesthetic volume marginally compromises postoperative analgesia during the first 24 hours.


Effect of antibodies to type 1 fimbriae on clearance of fimbriated Escherichia coli from the bloodstream of turkeys.
Young turkeys (n = 20) were inoculated IV with fimbriated, virulent Escherichia coli ECl (O78:K80: H9:F1). Blood samples were collected for bacterial quantitation at postinoculation minutes (PIM) 10, 20, 30, 40, 50, and 60. Immediately after the PIM 30 sampling, the turkeys were allotted into 4 groups (5 turkeys/group) and were injected IV with 1 of the following antisera: group 1, antibodies to F1 fimbriae (AF); group 2, antibodies to E coli O78 antigen (AO); group 3, antibodies to live, fimbriated (F1+) homologous E coli (ALEC); or group 4, normal turkey serum (NTS) collected from a healthy turkey. Compared with NTS, ALEC and AO caused a significant reduction in blood-borne E coli, whereas AF did not reduce bacterial numbers. In addition, 2 groups of 10 turkeys were inoculated IV with live, F1+ or nonfimbriated (F1-) E coli ECl. Numbers of viable bacteria were determined in blood samples and liver specimens collected 2 minutes after inoculation. Compared with F1- bacteria, significantly more F1+ bacteria were found in liver specimens and significantly fewer F1+ bacteria were found in blood samples. Results indicated that antibodies to F1 fimbriae do not enhance clearance of F1+ E coli from the bloodstream of turkeys probably because F1+ bacteria are selectively cleared by the liver, even without antibody.


[The course of dilated cardiomyopathy (long-term follow-up data)].
A long-term (1-8 years) prospective follow-up of 224 patients with dilated cardiomyopathy permitted identification of 3 variants of the disease run: benign (9.4%), protracted (51.8%) and malignant (38.8%). These variants are characterized by different gravity and time course changes in cardiovascular function and pumping capacity affections, in hypertrophy of the cardiac muscle, in incidence rate, time and causes of lethal outcomes.


Immunocytochemical analysis of the rat pineal gland using antisera generated against luteinizing hormone-releasing hormone (LHRH).
Seven different antisera generated against synthetic luteinizing hormone-releasing hormone (LHRH) were used to stain pineal gland and median eminence of the hypothalamus of normal rats. The LHRH immunogen complexes used to generate these antisera produced either N- or C-terminal directed antibodies or antisera that recognized both portions of the hormone molecule. Of these, four revealed LHRH-like immunoreactive material in the median eminence and pineal gland, two stained median eminence and not pineal, and one was positive in pineal only. These observations suggest that immunoreactive material in pineal gland may be an LHRH-like substance whose affinity characteristics are different from that of hypothalamic LHRH.


A Cut-and-Paste Approach to 3D Graphene-Oxide-Based Architectures.
Properly cut sheets can be converted into complex 3D structures by three basic operations including folding, bending, and pasting to render new functions. Folding and bending are extensively employed in crumpling, origami, and pop-up fabrications for 3D structures. Pasting joins different parts of a material together, and can create new geometries that are fundamentally unattainable by folding and bending. However, it has been much less explored, likely due to limited choice of weldable thin film materials and residue-free glues. Here it is shown that graphene oxide (GO) paper is one such suitable material. Stacked GO sheets can be readily loosened up and even redispersed in water, which upon drying, restack to form solid structures. Therefore, water can be utilized to heal local damage, glue separated pieces, and release internal stress in bent GO papers to fix their shapes. Complex and dynamic 3D GO architectures can thus be fabricated by a cut-and-paste approach, which is also applicable to GO-based hybrid with carbon nanotubes or clay sheets.


Calcium dependent regulation of brain and cardiac muscle adenylate cyclase.
The very close interdependence of Ca2+ and hormones in the overall metabolism of cyclic nucleotides has recently been emphasized by Cheung. Clearly the results presented here show that [Ca2+] in the physiological range (less than 10(-7) M to greater than 10(-6) M) has profound effects on the activity of adenylate cyclase from both brain and cardiac muscle. Whereas both brain and cardiac cyclase exhibit a Ca2+ dependent inhibition (perhaps mediated by calmodulin), only the brain cyclase is activated by Ca2+ via calmodulin. With both cyclases there is an inverse relationship between the inhibition of cyclase and the activation of calmodulin dependent (cAMP and cGMP) phosphodiesterase as a function of Ca2+ concentration. Because the IC50's for Ca2+ are the same in both heart and brain, the possibility exists that the Ca2+ inhibitory site of both cyclases is similar and perhaps identical. Considering the ability of Ca2+ to both stimulate and inhibit cyclase, one could imagine that in different species, tissues, or regions of the same tissue, there could exist multiple populations of cyclase, that is a cyclase which would only show Ca2+ dependent inhibition, Ca2+ dependent stimulation, or the biphasic response to Ca2+ (FIGURE 7). The fact that Ca2+ still regulates adenylate cyclase after various stimuli (histamine, NaF, etc.) suggests that Ca2+ may function to regulate the cyclase over shorter time periods (regardless of its state of stimulation) and that other affectors of cyclase (e.g., hormones) would serve to regulate the cyclase over longer time periods.


Perinatal outcome in 41 sets of triplets in Jordan.
Triplet births, which have increased greatly throughout the world in recent years, have a much greater risk of poor birth outcome than singleton births. The purpose of this study was to determine the perinatal outcome associated with triplet pregnancies and to compare abdominal delivery with vaginal delivery. We conducted a retrospective study of 41 sets of triplets born between January 1, 1994, and June 30, 1999, at the Princess Badee'a Teaching Hospital in Amman, Jordan. The primary outcome measures were perinatal mortality and early neonatal complications. Of these sets, 21 triplets were delivered vaginally and 20 triplets were delivered by cesarean section. The perinatal mortality rate was 260 per 1000 live births in this series, primarily due to respiratory distress syndrome. The perinatal deaths occurred to infants whose birthweights were primarily 500 to 1500 g (90.6%). Breech presentation was associated with a significantly higher perinatal mortality rate than vertex presentation (62.5% vs 37.5%). Cesarean delivery was associated with a higher perinatal mortality rate than vaginal delivery (30.0% vs 22.2%). These results suggested that cesarean delivery in triplets is not superior to vaginal delivery in terms of fetal and early neonatal outcome. The perinatal mortality rate was significantly higher than that in other recent series due to limited resources in Jordan.


Quasiparticle scattering in type-II Weyl semimetal MoTe2.
The electronic structure of type-II Weyl semimetal molybdenum ditelluride (MoTe2) is studied by using scanning tunneling microscopy and density functional theory calculations. Through measuring energy-dependent quasiparticle interference (QPI) patterns with a cryogenic scanning tunneling microscope, several characteristic features are found in the QPI patterns. Two of them arise from the Weyl semimetal nature; one is the topological Fermi arc surface state and the other can be assigned to be a Weyl point. The remaining structures are derived from the scatterings relevant to the bulk electronic states. The findings lead to further understanding of the topological electronic structure of type-II Weyl semimetal MoTe2.


Epidemiology, pathogenesis, and diagnosis of Aleutian disease caused by Aleutian mink disease virus: A literature review with a perspective of genomic breeding for disease control in American mink (Neogale vison).
Aleutian disease (AD) is a multi-systemic infectious disease in American mink (Neogale vison) caused by the Aleutian mink disease virus (AMDV). Commonly referred to as mink plasmacytosis, AD is an economically significant disease in mink-breeding countries. Aleutian disease mainly induces weight loss, lower fertility, and dropped pelt quality in adults and can result in acute interstitial pneumonia with high mortality rates in kits. In this review, we employed the scientific literature on AD over the last 70 years to discuss the historical and contemporary status of AD outbreaks and seroprevalence in mink farming countries. We also explained different forms of AD and the differences between the pathogenicity of the virus in kits and adults. The application of the available AD serological tests in AD control strategies was argued. We explained how selection programs could help AD control and proposed different approaches to selecting animals for building AD-tolerant herds. The advantages of genomic selection for AD tolerance over traditional breeding strategies were discussed in detail. We also explained how genomic selection could help AD control by selecting tolerant animals for the next generation based on genome-wide single nucleotide polymorphisms (SNP) data and the challenges of implementing genomic selection for AD tolerance in the mink industry. This review collected the information required for designing successful breeding programs for AD tolerance. Examples of the application of information are presented, and data gaps are highlighted. We showed that AD tolerance is necessary to be among the traits that animals are selected for in the mink industry.


Machine Learning Predicts Accurately Mycobacterium tuberculosis Drug Resistance From Whole Genome Sequencing Data.
Background: Tuberculosis disease, caused by Mycobacterium tuberculosis, is a major public health problem. The emergence of M. tuberculosis strains resistant to existing treatments threatens to derail control efforts. Resistance is mainly conferred by mutations in genes coding for drug targets or converting enzymes, but our knowledge of these mutations is incomplete. Whole genome sequencing (WGS) is an increasingly common approach to rapidly characterize isolates and identify mutations predicting antimicrobial resistance and thereby providing a diagnostic tool to assist clinical decision making. Methods: We applied machine learning approaches to 16,688 M. tuberculosis isolates that have undergone WGS and laboratory drug-susceptibility testing (DST) across 14 antituberculosis drugs, with 22.5% of samples being multidrug resistant and 2.1% being extensively drug resistant. We used non-parametric classification-tree and gradient-boosted-tree models to predict drug resistance and uncover any associated novel putative mutations. We fitted separate models for each drug, with and without "co-occurrent resistance" markers known to be causing resistance to drugs other than the one of interest. Predictive performance was measured using sensitivity, specificity, and the area under the receiver operating characteristic curve, assuming DST results as the gold standard. Results: The predictive performance was highest for resistance to first-line drugs, amikacin, kanamycin, ciprofloxacin, moxifloxacin, and multidrug-resistant tuberculosis (area under the receiver operating characteristic curve above 96%), and lowest for third-line drugs such as D-cycloserine and Para-aminosalisylic acid (area under the curve below 85%). The inclusion of co-occurrent resistance markers led to improved performance for some drugs and superior results when compared to similar models in other large-scale studies, which had smaller sample sizes. Overall, the gradient-boosted-tree models performed better than the classification-tree models. The mutation-rank analysis detected no new single nucleotide polymorphisms linked to drug resistance. Discordance between DST and genotypically inferred resistance may be explained by DST errors, novel rare mutations, hetero-resistance, and nongenomic drivers such as efflux-pump upregulation. Conclusion: Our work demonstrates the utility of machine learning as a flexible approach to drug resistance prediction that is able to accommodate a much larger number of predictors and to summarize their predictive ability, thus assisting clinical decision making and single nucleotide polymorphism detection in an era of increasing WGS data generation.


Zac1 plays a key role in the development of specific neuronal subsets in the mouse cerebellum.
The cerebellum is composed of a diverse array of neuronal subtypes. Here we have used a candidate approach to identify Zac1, a tumor suppressor gene encoding a zinc finger transcription factor, as a new player in the transcriptional network required for the development of a specific subset of cerebellar nuclei and a population of Golgi cells in the cerebellar cortex. We found that Zac1 has a complex expression profile in the developing cerebellum, including in two proliferating progenitor populations; the cerebellar ventricular zone and the external granular layer overlying posterior cerebellar lobules IX and X. Zac1 is also expressed in some postmitotic cerebellar neurons, including a subset of GABAergic interneurons in the medial cerebellar nuclei. Notably, GABAergic interneurons in the cerebellar nuclei are derived from the cerebellar ventricular zone, where Zac1 is also expressed, consistent with a lineage relationship between these two Zac1+ populations. Zac1 is also expressed in a small subset of cells in the posterior vermis, including some neurogranin-immunoreactive (NG+) Golgi cells, which, based on short-term birthdating, are derived from the EGL, where Zac1 is also expressed. However, Zac1+ cells and NG+ Golgi cells in the cerebellar cortex also display unique properties, as they are generated within different, albeit overlapping, time windows. Finally, consistent with the expression profile of Zac1, two conspicuous abnormalities were found in the cerebellum of Zac1 null mice: the medial cerebellar nuclei, and not the others, were significantly reduced in size; and the number of Golgi cells in cerebellar lobule IX was reduced by approximately 60% compared to wild-type littermates. The data presented here indicate that the tumor suppressor gene Zac1 is expressed in a complex fashion in the developing cerebellum, including in two dividing progenitor populations and in specific subsets of postmitotic neurons, including Golgi cells and GABAergic neurons in the medial nuclei, which require Zac1 for their differentiation. We thus conclude that Zac1 is a critical regulator of normal cerebellar development, adding a new transcriptional regulator to the growing list of factors involved in generating neuronal diversity in the developing cerebellum.