Sheep use preingestive cues as indicators of postingestive consequences to improve food learning.
Previous work has shown that herbivores successfully learn to associate food sensory characteristics with postingestive consequences when the conditioning procedure is simple, whereas this ability breaks down when the learning task is made more complex. We hypothesized that sensory characteristics could act as indicators of postingestive consequences and that the presence of preingestive cues would improve the food learning of sheep in situations varying in complexity. Sixteen sheep were subjected to a first conditioning phase to associate 2 flavors added to alfalfa hay with either a positive or a negative consequence, induced by intraruminal administration of starch (330 mg/g of DMI) or LiCl (5 mg/g of DMI). Sheep progressively decreased their choice of the flavored hay associated with the negative consequence (P < 0.05). This procedure provided sheep with experience with postingestive consequences associated with the different flavors. In a second conditioning phase, the experienced sheep and 16 naïve sheep were divided into groups of 8 and subjected to either a simple or a complex conditioning procedure [i.e., the 2 flavors were offered on separate days (simple conditioning) or simultaneously within a day (complex conditioning)]. The 2 flavors applied to grass hay were associated with either positive (starch, 330 mg/g of DMI) or negative (LiCl, 10 mg/g of DMI) consequences. As hypothesized, sheep in the simple conditioning group expressed a greater aversion to the flavored hay associated with the negative consequence than did those in the complex conditioning group (0.303 +/- 0.035 vs. 0.474 +/- 0.035 respectively; P < 0.01). Experienced sheep rejected the flavor associated with the negative consequence more strongly than did naïve sheep, regardless of the conditioning procedure (0.304 +/- 0.029 vs. 0.470 +/- 0.041 respectively; P < 0.05). The initial increased preference for aniseed (0.80 +/- 0.04), however, greatly influenced food learning because sheep negatively conditioned on this flavor expressed less avoidance than those negatively conditioned on orange (0.53 +/- 0.04 vs. 0.25 +/- 0.03 respectively; P < 0.01). In conclusion, the simultaneous scenario was actually perceived as complex by all sheep, and experienced sheep were more efficient in food learning than naïve sheep in both the simple and complex learning contexts. The sheep were thus able to generalize the association between sensory cues and postingestive consequences, especially in a complex environment, and then to use these sensory cues as indicators of postingestive consequences.


The inhibitory effect of phosphorylase a on the activation of glycogen synthase depends on the type of synthase phosphatase.
The activity of glycogen synthase phosphatase in rat liver stems from the co-operation of two proteins, a cytosolic S-component and a glycogen-bound G-component. It is shown that both components possess synthase phosphatase activity. The G-component was partially purified from the enzyme-glycogen complex. Dissociative treatments, which increase the activity of phosphorylase phosphatase manyfold, substantially decrease the synthase phosphatase activity of the purified G-component. The specific inhibition of glycogen synthase phosphatase by phosphorylase a, originally observed in crude liver extracts, was investigated with purified liver synthase b and purified phosphorylase a. Synthase phosphatase is strongly inhibited, whether present in a dilute liver extract, in an isolated enzyme-glycogen complex, or as G-component purified therefrom. In contrast, the cytosolic S-component is insensitive to phosphorylase a. The activation of glycogen synthase in crude extracts of skeletal muscle is not affected by phosphorylase a from muscle or liver. Consequently we have studied the dephosphorylation of purified muscle glycogen synthase, previously phosphorylated with any of three protein kinases. Phosphorylase a strongly inhibits the dephosphorylation by the hepatic G-component, but not by the hepatic S-component or by a muscle extract. These observations show that the inhibitory effect of phosphorylase a on the activation of glycogen synthase depends on the type of synthase phosphatase.


Cholera dynamics: lessons from an epidemic.
Cholera is a severe diarrhoeal disease that spreads rapidly and affects millions of people each year, resulting in tens of thousands of deaths. The disease is caused by Vibrio cholerae O1 and is characterized by watery diarrhoea that can be lethal if not properly treated. Cholera had not been reported in South America from the late 1800s until 1991, when it was introduced in Peru, wreaking havoc in one of the biggest epidemics reported to date. Within a year, the disease had spread to most of the Latin American region, resulting in millions of cases and thousands of deaths in all affected countries. Despite its aggressive entry, cholera virtually disappeared from the continent after 1999. The progression of the entire epidemic was well documented, making it an ideal model to understand cholera dynamics. In this review, we highlight how the synergy of socioeconomic, political and ecological factors led to the emergence, rapid spread and eventual disappearance of cholera in Latin America. We discuss how measures implemented during the cholera epidemic drastically changed its course and continental dynamics. Finally, we synthesize our findings and highlight potential lessons that can be learned for efficient and standardized cholera management programmes during future outbreaks in non-endemic areas.


Cannabis and anxiety and depression in young adults: a large prospective study.
To examine whether age of first use or frequency of use of cannabis is associated with anxiety and depression (AD) in young adults, independent of known potential confounders, including the use of other illicit drugs. A cohort of 3,239 Australian young adults was followed from birth to the age of 21 when data on AD were obtained from sample members along with information on their use of cannabis at 21 years. Potential confounding factors were prospectively measured when the child was born and at 14 years. After controlling for confounding factors, those who started using cannabis before age 15 years and used it frequently at 21 years were more likely to report symptoms of AD in early adulthood (odds ratio 3.4; 95% CI 1.9-6.1). This association was of similar magnitude for those who had only used cannabis and those who reported having used cannabis and other illicit drugs. The relationship between early-onset and frequent use of cannabis and symptoms of AD is independent of individual and family backgrounds. Frequent cannabis use is associated with increased AD in young adults independently of whether the person also uses other illicit drugs.


Suspected insulin anaphylaxis and literature review.
Insulin allergy is a well-documented complication of insulin therapy. A 67-year-old man presented with symptoms suggestive of insulin anaphylaxis. In an attempt to allow him to continue insulin therapy, he underwent a desensitization protocol. During the protocol, he again experienced symptoms suggestive of anaphylaxis. An analysis of his case is presented in the context of current literature. All physicians treating patients with insulin should be aware of this serious complication.


Prevalence of food insecurity in people with major depression, bipolar disorder, and schizophrenia and related psychoses: A systematic review and meta-analysis.
People with severe mental illness (SMI), such as major depression, bipolar disorder, and schizophrenia, experience numerous risk factors that may predispose them to food insecurity; however, the prevalence of food insecurity and its effects on health are under-researched in this population group. This systematic review and meta-analysis aimed to describe the prevalence and correlates of food insecurity in people with SMI. A comprehensive electronic search was conducted up to March 2021. Random effects meta-analysis was employed to determine the prevalence of food insecurity in SMI, and odds ratio (OR) of food insecurity in people with SMI compared to non-psychiatric controls/general population. Twenty-nine unique datasets (31 publications) were included. Prevalence estimate of food insecurity in people with SMI was 40% (95% CI 29-52%, I2 = 99.7%, N = 27). People with SMI were 2.71 (95% CI 1.72-3.25) times more likely to report food insecurity than the comparator group (Z = 11.09, p < 0.001, I2 = 95%, N = 23). The odds of food insecurity in SMI were higher in high/high-middle income countries compared to low/low-middle income countries, likely due to the high food insecurity rates in the general population of lower income countries. There was no difference in food insecurity rates by diagnosis. Food insecurity should be a consideration for health professionals working with community-dwelling people with SMI.


Core 2 branching beta1,6-N-acetylglucosaminyltransferase and high endothelial cell N-acetylglucosamine-6-sulfotransferase exert differential control over B- and T-lymphocyte homing to peripheral lymph nodes.
Blood-borne lymphocyte trafficking to peripheral lymph nodes (PLNs) depends on the successful initiation of rolling interactions mediated by L-selectin binding to sialomucin ligands in high endothelial venules (HEVs). Biochemical analysis of purified L-selectin ligands has identified posttranslational modifications mediated by Core2GlcNAcT-I and high endothelial cell GlcNAc-6-sulfotransferase (HECGlcNAc6ST). Consequently, lymphocyte migration to PLNs of C2GlcNAcT-I(-/-) and HEC-GlcNAc6ST(-/-) mice was reduced; however, B-cell homing was more severely compromised than T-cell migration. Accordingly, intravital microscopy (IVM) of PLN HEVs revealed a defect in B-cell tethering and increased rolling velocity (V(roll)) in C2GlcNAcT-I(-/-) mice that was more pronounced than it was for T cells. By contrast, B- and T-cell tethering was normal in HEC-GlcNAc6ST(-/-) HEVs, but V(roll) was accelerated, especially for B cells. The increased sensitivity of B cells to glycan deficiencies was caused by lower expression levels of L-selectin; L-selectin(+/-) T cells expressing L-selectin levels equivalent to those of B cells exhibited intravascular behavior similar to that of B cells. These results demonstrate distinct functions for C2GlcNAcT-I and HEC-GlcNAc6ST in the differential elaboration of HEV glycoproteins that set a threshold for the amount of L-selectin needed for lymphocyte homing.


[Principles of the structural organization of the cetacean neocortex].
The investigations performed demonstrate that morphologically cetacean neocortex is organized on the same principles as that of other mammals, as it has six layer structure, layer IV including. The results of investigations done for many years gave the possibility to establish quantitative characteristics of neocortex in different zoological cetacean genera, such as volume of neocortex, total number of its nervous cells (for dolphin the data were obtained for the first time), and clarify the problem fully enough. It became clear that morphologically dolphins belong to different genera. Three quantitative gradations of morphological enrichment of the dolphin neocortex are demonstrated. The only genus of Tursiops (bottle-nosed) can be compared with man, as to on what neuromorphological criteria it is behind, equal and surpasses the man. Representatives of the two other genera investigated, according to all neuromorphological indices studied, are far behind the man.


Genomic alterations and gene expression in primary diffuse large B-cell lymphomas of immune-privileged sites: the importance of apoptosis and immunomodulatory pathways.
Primary diffuse large B-cell lymphomas of different immune-privileged sites (IP-DLBCLs) share many clinical and biological features, such as a relatively poor prognosis, preferential dissemination to other immune-privileged sites, and deletion of the HLA region, which suggests that IP-DLBCL represents a separate entity. To further investigate the nature of IP-DLBCL, we investigated site-specific genomic aberrations in 16 testicular, nine central nervous system (CNS), and 15 nodal DLBCLs using array CGH. We also determined minimal common regions of gain and loss. Using robust algorithms including multiple testing procedures and the ACE-it script, which is specifically designed for this task, the array CGH data were combined with gene expression data to explore pathways deregulated by chromosomal aberrations. Loss of 6p21.32-p25.3, including the HLA genes, was associated with both types of IP-DLBCL, whereas gain of 2p16.1-p25.3 was associated with nodal DLBCL. Gain of 12q15-q21.1 and 12q24.32-q24.33 was associated with CNS DLBCL and gain of 19q13.12-q13.43 with testicular DLBCL. Analysis of candidate genes in site-specific regions and minimal common regions revealed two major groups of genes: one involved in the immune response, including regulation of HLA expression, and the other involved in apoptosis, including the p53 pathway. Many of these genes were also involved in homozygous deletions or high-level gains. The presence of both shared and site-specific aberrations in CNS and testicular DLBCLs underlines the concept of IP-DLBCL but also indicates that IP-DLBCLs of the CNS and testis do not form a single entity. The observed aberrations emphasize the importance of the deregulation of anti-tumour immune response and apoptosis pathways.


Cyclic Cushing's syndrome caused by neuroendocrine tumor: a case report.
Cushing's syndrome (CS) is a clinical syndrome characterized by hypercortisolemia. Cyclic Cushing's syndrome (CCS), which exhibits a periodic or irregular increasing pattern in cortisol, is a rare type of Cushing's syndrome. A 37-year-old man came to our hospital because of repeated dizzy spells, weakness and hypercortisolemia lasting two weeks. Endocrinological examinations indicated CCS with periodic and intermittent increases in cortisol. Enhanced computed tomography (CT) revealed space occupying lesions on the upper lobe of left lung, and biopsy eventually proved that these were pulmonary carcinoid tumors with ectopic ACTH secretion, which was subsequently manifested a Cushing's syndrome. PET-CT, ultrasound and biopsy of the thyroid gland indicated bilateral thyroid papillary carcinoma. CT scan showed bilateral nodular hyperplasia of the adrenal gland. Enhanced magnetic resonance imaging (MRI) confirmed that the high signal disappeared on the posterior lobe of the pituitary gland and that the pituitary stalk shifted left, which was suspected to be non-functional pituitary microadenoma. The patient underwent surgery involving resection of the left upper pulmonary lobe and the mediastinal lymph node around the hilus pulmonis, which resulted in complete remission of CCS. The patient then chose elective surgery for the thyroid papillary carcinoma. An analysis of the patient's genomic DNA identified a novel mutation in PDE11A: c.2032 (exon 12) G > A, which is associated with primary pigmented nodular adrenocortical disease (PPNAD). This is a novel mutation which has been no previous public clinical report on this mutation as it relates to this disease.


The effects of situations and situation transitions on maternal and infant behavior.
Mothers and their firstborn year-old infants (30 girls, 30 boys) were observed during two counterbalanced situations. In the noninteraction situation, mothers were not to initiate interaction; during interaction mothers were free to interact. Maternal and infant behavior were dependent upon both the situation examined and how it compared to the previous situation. During interaction, mothers directed more behavior toward their infants than during noninteraction; this effect was intensified when noninteraction preceded interaction. Infant looking and proximity-seeking were greater during interaction than during noninteraction, while the reverse was true for touching and vocalization. A decrease in the level of interaction resulted in increases in infant behavior, while the reverse transition was associated with a more variable pattern. Two models describe the interrelationships between changes in maternal and infant behavior. In a reciprocal system, increases by the mother in a given behavior, or in the general interactive level are related to decreases in infant behavior, and vice versa. In an incremental system, any change in maternal behavior is met by increases in infant behavior.


Stereospecific in vivo N-methylation of nicotine in the guinea pig.
R-(+)-[3H-N'-CH3]N-methylnicotinium ion has been identified as a urinary metabolite of ip administered R-(+)-[3H-N'-CH3]nicotine in the guinea pig. Under similar conditions, S-(-)-[3H-N'-CH3]nicotine is not converted to the corresponding N-methylated metabolite. R-(+)-N-methylnicotinium salt was isolated from the urine of guinea pigs that had been chronically dosed ip with R-(+)-nicotine. The identification and stereochemical analysis of this metabolite were carried out using analytical and preparative cation exchange high pressure liquid chromatography and chemical reduction followed by GLC-mass spectrometric analysis and 1H NMR spectroscopy. The results show that nicotine is stereospecifically biotransformed into an N-methylated urinary metabolite in the guinea pig.


Functional determination of McN-A-343 affinity for M1 muscarinic receptors.
The affinity (KA) of 4-(m-chlorophenyl-carbamoyloxy)-2-butynyltrimethylammonium chloride (McN-A-343), acting as an agonist of M1 muscarinic receptors, has been estimated by means of fractional receptor inactivation, employing the irreversible muscarinic antagonist propylbenzylcholine mustard. Two M1-mediated responses elicited by McN-A-343 were studied: relaxation of the isolated rat duodenum and inhibition of twitch contractions in rabbit was deferens. A comparison was made with the affinity of McN-A-343 as an antagonist (KB) of acetylcholine-induced contraction in rat duodenum. Results showed that McN-A-343 displayed similar affinities as an agonist and as an antagonist: -log KA were 4.68 and 5.17 in duodenum and vas deferens, respectively, vs. -log KB of 4.96 in duodenum, indicating that the ability of McN-A-343 to selectively stimulate M1 receptors is not based on a greater affinity for this subtype. In both preparations examined, McN-A-343 reached maximum effect through occupation of a fraction of the total available receptors (approximately 30% in duodenum, approximately 80% in vas deferens), implying that occupation of M1 receptors is translated into effect in a highly efficient way.


Radiofrequency Thermal Ablation versus Bipolar Electrocautery for the Treatment of Inferior Turbinate Hypertrophy: Comparison of Efficacy and Postoperative Morbidity.
Introduction Numerous surgical methods are used to treat nasal obstruction due to inferior turbinate hypertrophy. The primary goal of the therapy is to maximize the nasal airway for as extended a period of time as possible while minimizing therapeutic complications. Objectives The aim of this study was to assess the effects of radiofrequency thermal ablation (RFTA) and bipolar electrocautery (BEC) on the removal of nasal obstruction in patients with inferior turbinate hypertrophy and on nasal mucociliary clearance (MCC). Patients in both groups were also evaluated in terms of postoperative morbidity. Methods We compared the outcomes of two groups of patients: those treated with RFTA (n = 23) and those who underwent BEC (n = 20). Nasal obstruction was graded using a visual analog scale (VAS) and MCC was measured using a saccharin clearance test. Both measurements were performed before and 2 months after treatment. Results Pre- and postoperative VAS scores showed significant improvement for both groups. However, MCC results did not significantly differ between two groups. Neither edema nor crust formation persisted for more than 1 week in any patients. Conclusion Submucosal cauterization with preservation of the nasal mucosa and periosteum is as effective and safe as RFTA and should be considered when planning inferior turbinate interventions.


Hippocampal Electrical Activity and Behaviour in Rabbits during Adaptation to a Novel Semi-Natural Environment.
The aim of the present experiment was to show possible modifications in behaviour and hippocampal electrical activity during adaptation to a novel semi-natural environment. Rabbits with chronic implanted electrodes in the hippocampus were introduced into a large, natural, open-air enclosure. Their spontaneous behaviour was studied for 3 days. Electrical activity was recorded by telemetry, stored with behaviour on a video-tape, and then analysed. Behaviour was divided into categories and variables. The categories were: activity directed towards the environment (including exploratory movements, scanning, marking, etc.) and self-directed activities (such as eating, drinking, self-grooming, etc.). Exploratory elements were observed and classified according to a spatial and a temporal criterion. The rabbits showed a progressive adaptation to the environment with quantitative and qualitative reduction in exploration and an increment in quiet immobility and inspective activities. Three EEG patterns were recognized from autocorrelation and spectral analysis: high rhythmicity theta (HRSA) with specific relationship to voluntary movements, such as exploration and running; low rhythmicity theta (LRSA) in signalling activities or self-directed behaviour and during postural and reactive immobility; and irregular activity (IA) sometimes seen in the same immobile behaviour. The results show that some behaviours (exploration, alert and postural immobility, self-directed activity) are correlated to specific patterns of electrical activity in the hippocampus and, as exploratory behaviour decreased with time, there was a concomitant increase in EEG frequency.


Autophagy and metastasis: another double-edged sword.
In order to metastasize, tumor cells must adapt to untoward, stressful microenvironments as they disseminate into the systemic circulation and colonize distant organ sites. Autophagy, a tightly regulated lysosomal self-digestion process that is upregulated during cellular stress, has been demonstrated to suppress primary tumor formation, but how autophagy influences metastasis remains unknown. Autophagy may inhibit metastasis by promoting antitumor inflammatory responses or by restricting the expansion of dormant tumor cells into macrometastases. Conversely, self-eating may promote metastasis by enhancing tumor cell fitness in response to environmental stresses, such as anoikis, during metastatic progression. Because autophagy is titratable, it may serve both prometastatic and antimetastatic functions depending on the contextual demands placed on tumor cells throughout the metastatic process.


A retinoid X receptor (RXR)-selective retinoid reveals that RXR-alpha is potentially a therapeutic target in breast cancer cell lines, and that it potentiates antiproliferative and apoptotic responses to peroxisome proliferator-activated receptor ligands.
Certain lipids have been shown to be ligands for a subgroup of the nuclear hormone receptor superfamily known as the peroxisome proliferator-activated receptors (PPARs). Ligands for these transcription factors have been used in experimental cancer therapies. PPARs heterodimerize and bind DNA with retinoid X receptors (RXRs), which have homology to other members of the nuclear receptor superfamily. Retinoids have been found to be effective in treating many types of cancer. However, many breast cancers become resistant to the chemotherapeutic effects of these drugs. Recently, RXR-selective ligands were discovered that inhibited proliferation of all-trans retinoic acid resistant breast cancer cells in vitro and caused regression of the disease in animal models. There are few published studies on the efficacy of combined therapy using PPAR and RXR ligands for breast cancer prevention or treatment. We determined the effects of selective PPAR and RXR ligands on established human breast cancer cell lines in vitro. PPAR-alpha and PPAR-gamma ligands induced apoptotic and antiproliferative responses in human breast cancer cell lines, respectively, which were associated with specific changes in gene expression. These responses were potentiated by the RXR-selective ligand AGN194204. Interestingly, RXR-alpha-overexpressing retinoic acid resistant breast cancer cell lines were more sensitive to the effects of the RXR-selective compound. RXR-selective retinoids can potentiate the antiproliferative and apoptotic responses of breast cancer cell lines to PPAR ligands.


Microfracture: Its History and Experience of the Developing Surgeon.
Development and implementation of the microfracture technique began in the early 1980s. The surgical goal was to produce "microfractures" in the subchondral bone perpendicular to the surface and to be able to reach all areas of the joint with the instruments. The microfracture technique has been demonstrated to be an effective arthroscopic treatment for full-thickness chondral lesions and joints with degenerative lesions. This technique is cost effective, technically not complicated, has an extremely low rate of associated patient morbidity, and leaves options for further treatment. Patient compliance with rehabilitation, knee alignment, and the depth of the cartilage rim surrounding the lesion are a few of the factors that can affect the outcomes following microfracture.


Population pharmacokinetics of oxcarbazepine and its monohydroxy derivative in epileptic children.
Oxcarbazepine is an antiepileptic drug with an activity mostly due to its monohydroxy derivative metabolite (MHD). A parent-metabolite population pharmacokinetic model in children was developed to evaluate the consistency between the recommended paediatric doses and the reference range for trough concentration (Ctrough ) of MHD (3-35 mg l-1 ). A total of 279 plasma samples were obtained from 31 epileptic children (age 2-12 years) after a single dose of oxcarbazepine. Concentration-time data were analysed with Monolix 4.3.2. The probability to obtain Ctrough between 3-35 mg l-1 was determined by Monte Carlo simulations for doses ranging from 10 to 90 mg kg-1 day-1 . A parent-metabolite model with two compartments for oxcarbazepine and one compartment for MHD best described the data. Typical values for oxcarbazepine clearance, central and peripheral distribution volume and distribution clearance were 140 l h-1 70 kg-1 , 337 l 70 kg-1 , 60.7 l and 62.5 l h-1 , respectively. Typical values for MHD clearance and distribution volume were 4.11 l h-1 70 kg-1 and 54.8 l 70 kg-1 respectively. Clearances and distribution volumes of oxcarbazepine and MHD were related to body weight via empirical allometric models. Enzyme-inducing antiepileptic drugs (EIAEDs) increased MHD clearance by 29.3%. Fifty-kg children without EIAEDs may need 20-30 mg kg-1 day-1 instead of the recommended target maintenance dose (30-45 mg kg-1 day-1 ) to obtain Ctrough within the reference range. By contrast, 10-kg children with EIAEDs would need 90 mg kg-1 day-1 instead of the maximum recommended dose of 60 mg kg-1 day-1 . This population pharmacokinetic model of oxcarbazepine supports current dose recommendations, except for 10-kg children with concomitant EIAEDs and 50-kg children without EIAEDs.


Effect of epithelial-specific MyD88 signaling pathway on airway inflammatory response to organic dust exposure.
The Toll-like receptor (TLR) adaptor protein MyD88 is integral to airway inflammatory response to microbial-enriched organic dust extract (ODE) exposures. ODE-induced airway neutrophil influx and release of pro-inflammatory cytokines was essentially abrogated in global MyD88-deficient mice, yet these mice demonstrate an increase in airway epithelial cell mucin expression. To further elucidate the role of MyD88-dependent responses specific to lung airway epithelial cells in response to ODE in vivo, the surfactant protein C protein (SPC) Cre+ embryologic expressing airway epithelial cells floxed for MyD88 to disrupt MyD88 signaling were utilized. The inducible club cell secretory protein (CCSP) Cre+, MyD88 floxed, were also developed. Using an established protocol, mice were intranasally instilled with ODE or saline once or daily up to 3 weeks. Mice with MyD88-deficient SPC+ lung epithelial cells exhibited decreased neutrophil influx following ODE exposure once and repetitively for 1 week without modulation of classic pro-inflammatory mediators including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and neutrophil chemoattractants. This protective response was lost after 3 weeks of repetitive exposure. ODE-induced Muc5ac mucin expression at 1 week was also reduced in MyD88-deficient SPC+ cells. Acute ODE-induced IL-33 was reduced in MyD88-deficient SPC+ cells whereas serum IgE levels were increased at one week. In contrast, mice with inducible MyD88-deficient CCSP+ airway epithelial cells demonstrated no significant difference in experimental indices following ODE exposure. Collectively, these findings suggest that MyD88-dependent signaling targeted to all airway epithelial cells plays an important role in mediating neutrophil influx and mucin production in response to acute organic dust exposures.


The costs of children with sickle cell anemia: preparing for managed care.
To anticipate the clinical challenges and financial risks facing physicians and managed care organizations who care for children with chronic illnesses, such as sickle cell anemia (SCA), under capitated managed care arrangements. A cross-sectional study based on claims data from the Washington State Medicaid Program (WSMP) and the Federal Employees Health Benefits Program (FEP). Expenditure patterns were compared for children 18 years of age or younger for whom a claim with a diagnosis of SCA was submitted and paid in the State of Washington during fiscal year 1993 (FY1993) or by the FEP during FY1992 to expenditure patterns for all children. Children with SCA had mean expenditures 8.8 times the mean expenditures for all children in WSMP. There was wide variation in the annual expenditures among children with SCA; the most expensive 10% of children accounted for 56% of total expenditures. Ninety-seven percent of the expenditures were concentrated in four broad categories: 72% for inpatient care, 11% for outpatient care, 11% for physician payments, and 3% for prescription drugs. Examination of expenditure and utilization patterns for children with sickle cell anemia enrolled in the FEP yielded similar results. Unless managed care organizations and capitated pediatricians receive payment rates that reflect the higher expected expenditures of caring for these children, access to and quality of care may suffer. Analyses of practice guidelines and utilization patterns suggest that newborn screening, regular access to specialty facilities, and comprehensive education programs are critical areas that are vulnerable to reductions under capitation.


Serum bactericidal activity of rifampin in combination with other antimicrobial agents against Staphylococcus aureus.
Because rifampin-resistant strains of Staphylococcus aureus emerge during monotherapy with this drug, a search was made for potentially useful companion drugs. Bactericidal titers of spiked serum were determined, and time kill studies were performed for 10 strains of methicillin-susceptible S. aureus. We tested rifampin in combination with nafcillin, vancomycin, clindamycin, pefloxacin, ciprofloxacin, trimethoprim, teicoplanin, or erythromycin. The bactericidal activity of nafcillin, vancomycin, and teicoplanin was significantly reduced (P less than 0.05) when rifampin was added to the drug regimen. In contrast, the addition of rifampin to clindamycin or erythromycin significantly increased bactericidal activity as measured by both bactericidal titers in serum and 6-h killing rates (P less than 0.02). Bactericidal activity in serum was also increased by the addition of rifampin to trimethoprim, but rifampin-resistant strains emerged with this combination.


Critical assessment of alignment procedures for LC-MS proteomics and metabolomics measurements.
Liquid chromatography coupled to mass spectrometry (LC-MS) has become a prominent tool for the analysis of complex proteomics and metabolomics samples. In many applications multiple LC-MS measurements need to be compared, e. g. to improve reliability or to combine results from different samples in a statistical comparative analysis. As in all physical experiments, LC-MS data are affected by uncertainties, and variability of retention time is encountered in all data sets. It is therefore necessary to estimate and correct the underlying distortions of the retention time axis to search for corresponding compounds in different samples. To this end, a variety of so-called LC-MS map alignment algorithms have been developed during the last four years. Most of these approaches are well documented, but they are usually evaluated on very specific samples only. So far, no publication has been assessing different alignment algorithms using a standard LC-MS sample along with commonly used quality criteria. We propose two LC-MS proteomics as well as two LC-MS metabolomics data sets that represent typical alignment scenarios. Furthermore, we introduce a new quality measure for the evaluation of LC-MS alignment algorithms. Using the four data sets to compare six freely available alignment algorithms proposed for the alignment of metabolomics and proteomics LC-MS measurements, we found significant differences with respect to alignment quality, running time, and usability in general. The multitude of available alignment methods necessitates the generation of standard data sets and quality measures that allow users as well as developers to benchmark and compare their map alignment tools on a fair basis. Our study represents a first step in this direction. Currently, the installation and evaluation of the "correct" parameter settings can be quite a time-consuming task, and the success of a particular method is still highly dependent on the experience of the user. Therefore, we propose to continue and extend this type of study to a community-wide competition. All data as well as our evaluation scripts are available at http://msbi.ipb-halle.de/msbi/caap.


Psychiatric medication-induced obesity: a review.
A majority of psychiatric medications are known to generate weight gain and ultimately obesity in some patients. There is much speculation about the prevalence of weight gain and the degree of weight gain during acute and longitudinal treatment with these agents. There is newer literature looking at the aetiology of this weight gain and the potential treatments being used to alleviate this side-effect. We found solid evidence that weight gain is often associated with the mood stabilizers, and antipsychotics and antidepressants. Only few weight neutral or weight loss producing psychotropics are available, and weight gain, outside of an immediate side-effect, may generate secondary side-effects and medical comorbidity. Weight gain may cause hypertension, diabetes, osteoarthritis, sedentary lifestyle, coronary artery disease, etc. Given the likelihood of inducing weight gain with psychotropic medications and the longitudinal impact on physical health, a thorough literature review is warranted to determine the epidemiology, aetiology and treatment options of psychotropic-induced weight gain.


A new epistasis group for the repair of DNA damage in bacteriophage T4: replication repair.
The gene 32 mutation amA453 sensitizes bacteriophage T4 to the lethal effects of ultraviolet (UV) irradiation, methyl methanesulfonate and angelicin-mediated photodynamic irradiation when treated particles are plated on amber-suppressing host cells. The increased UV sensitivity caused by amA453 is additive to that caused by mutations in both the T4 excision repair (denV) and recombination repair (uvsWXY) systems, suggesting the operation of a third kind of repair system. The mutation uvs79, with many similarities to amA453 but mapping in gene 41, is largely epistatic to amA453. The mutation mms1, also with many similarities to amA453, maps close to amA453 within gene 32 and is largely epistatic to uvs79. Neither amA453 nor uvs79 affect the ratio of UV-induced mutational to lethal hits, nor does amA453 affect spontaneous or UV-enhanced recombination frequencies. Gene 32 encodes the major T4 ssDNA-binding protein (the scaffolding of DNA replication) and gene 41 encodes a DNA helicase, both being required for T4 DNA replication. We conclude that a third repair process operates in phage T4 and suggest that it acts during rather than before or after DNA replication.


Focusing and imaging of a virtual all-optical tunable terahertz Fresnel zone plate.
A virtual all-optical tunable terahertz Fresnel zone plate is achieved utilizing the localized distribution of the transient electron plasma on a silicon wafer. Its focusing and imaging performance are experimentally demonstrated. Experimental results show that the effect of the virtual zone plate is the same as an actual one. Adjusting the spatial pattern of the electron plasma, the central wavelength and the focal length of the virtual zone plate can be all-optically dynamically steered. The research is a significant step to the development of tunable optical imaging elements.


Effects of prenatal and chronic undernutrition on aging and survival in rats.
The effect on aging and survival of mild (2/3 of ad lib) chronic prenatal and postnatal undernutrition over 10 generations, or prenatal undernutrition only has been studied in rats. Both regimes of undernutrition used decreased body weights of animals as compared to the controls. Both undernourished groups had lower cholesterol levels and tumor incidence than the controls. The effects on survival depended not only on the time period of undernutrition but also on the age of the animals examined: chronic undernutrition resulted in lower survival of young adult animals (4-8 months) but higher survival than the controls in old age. Prenatal undernutrition only had no effect on young adult animals, gave higher survival than the controls at ages 8-18 months but considerably lower survival than controls in the old age: the underdevelopment of vital organs whose cells proliferate only before birth might have been the cause of such decreased longevity.


Comparison of Two Validation Nutrition Tools in Hospitalized Elderly: Full Mini Nutritional Assessment and Short-form Mini Nutritional Assessment.
The aim was to determine validity, reliability, and cutoff of full-mini nutritional assessment (MNA) and MNA-short form (SF) also which one was better for the screening of malnutrition in the Iranian hospitalized elderly. In this cross-sectional validation study, 96 hospitalized elderly ≥60 years selected from two hospitals in Tehran. Anthropometric measures (body mass index [BMI], mid-arm circumference [MAC], calf circumference [CC], abdomen, and waist skinfold thickness) and laboratory tests (albumin and hemoglobin levels, and red blood cell count were performed. Nutrition tools (full-MNA and MNA-SF), cognition tool (mini-mental state examination, depression scale (Geriatric Depression Scale15 and activities of daily living (ADL) index (Modified Barthel-ADL) were administered. The full-MNA scores were significantly correlated to measures of MAC, BMI, waist, and CC. The MNS-SF scores were significantly related to measures of MAC, waist, and CC. Serum albumin showed a poor correlation with both tools. At cutoff 24 in full-MNA had a sensitivity 75% and specificity 77.8% and the MNA-SF considered 62.5% sensitivity and 65.3% specificity at cutoff 10.50 to detect well-nourished from malnourished subjects. The internal consistencies of both tools were >90%. In exploratory factor analysis, six components found for full-MNA and two components for MNA-SF. Known group validity of full-MNA was reflected significant differences between geriatric patients with expected higher full-MNA scores and patients with expected lower scores (BMI ≥24 vs. BMI <24 or bed ulcer or assisted food intake). It seems the Persian version of full-MNA is more appropriate in comparison to MNA-SF for screening malnutrition in the Iranian hospitalized elderly patients.


Rest tremor in rhesus monkeys with MPTP-induced parkinsonism.
Rest tremor (RTr) is a typical feature of Parkinson's diseases (PD). Animal models of PD presenting with RTr are indispensable for understanding the pathophysiology of human RTr and the development of new therapeutic agents. In this report we studied the occurrence of tremor on rhesus monkeys rendered parkinsonian by an intracarotid (ICA) infusion followed by 2-4 iv. doses of n-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The animals' parkinsonism was assessed using a rating scale, activity monitors and a novel tremor monitor. The animals manifested bilateral parkinsonism with more severe clinical signs on the side of the body contralateral to the ICA infusion. The RTr in these animals had a mean peak frequency of 7.9 Hz (S.E.: 0.12), and a mean amplitude of 5.1/d/s/rtz (S.E.: 0.69). Substantial reduction in RTr amplitude (80.4%) was observed after oral L-DOPA administration. Our results suggest that: 1) RTr is present after the combined administration of ICA and iv. MPTP. 2) The mean RTr frequency in rhesus monkeys may be higher than in parkinsonian patients. However, as in PD, RTr frequency in the monkey was maintained within a narrow band width. 3) As in PD, L-DOPA administration to MPTP-treated monkeys reduced the amplitude of RTr and improved the parkinsonian features. Monitoring and quantifying the RTr in the MPTP-parkinsonian monkeys provide an objective, non invasive way to measure the outcome of therapeutic interventions and, further support the concept that loss of dopaminergic innervation contributes to the occurrence of RTr.


Synthetic Methods Driven by the Photoactivity of Electron Donor-Acceptor Complexes.
The association of an electron-rich substrate with an electron-accepting molecule can generate a new molecular aggregate in the ground state, called an electron donor-acceptor (EDA) complex. Even when the two precursors do not absorb visible light, the resulting EDA complex often does. In 1952, Mulliken proposed a quantum-mechanical theory to rationalize the formation of such colored EDA complexes. However, and besides a few pioneering studies in the 20th century, it is only in the past few years that the EDA complex photochemistry has been recognized as a powerful strategy for expanding the potential of visible-light-driven radical synthetic chemistry. Here, we explain why this photochemical synthetic approach was overlooked for so long. We critically discuss the historical context, scientific reasons, serendipitous observations, and landmark discoveries that were essential for progress in the field. We also outline future directions and identify the key advances that are needed to fully exploit the potential of the EDA complex photochemistry.


Characterization and aging study of currency ink and currency canine training aids using headspace SPME/GC-MS.
Solid-phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) was used to characterize the volatile components associated with U.S. currency, U.S. currency inks, and Canadian currency. Compounds that can be attributed to the ink-curing process include series of straight-chain aldehydes, alkenals, acids, alcohols, and ketones and a series of lactones and 2-alkyl furans. Solvent compounds include naphthenic and paraffinic hydrocarbons with a profile typical of petroleum products, alkyl cyclohexanes, various ethylene glycol alkyl ethers, and traces of chlorinated solvents. Trace levels of 2-phenoxyethanol, a solvent often used in ink formulations, were also detected. Environmental contaminants, those compounds found in circulating currency but not in currency ink, include 2,2'-diethyl-1,1'-biphenyl, methyl benzoate and salicylate, menthol, limonene, dimethyl and diethyl phthalate, and ionol. Not including simple hydrocarbons, over 100 compounds were identified in the headspace of currency-related samples.


Steroid dimers-in vitro cytotoxic and antimicrobial activities.
The in vitro cytotoxic activity of previously synthesized steroid dimers with different spacer group (sulfide, trithiolane ring or phosphorotrithioate) and the substituent at C-17 position was tested for their possible effects against following human tumor cell lines: cervical adenocarcinoma (HeLa), chronic myelogenous leukemia (K562) and two human breast cancer cell lines (MDA-MB-361 and MDA-MB-453). These compounds, applied at micromolar concentrations, exhibited cytotoxic activity of different intensity (compared with cisplatin as a control), modality and selectivity in these malignant cell lines. The best activity against all four cell cancer lines was exhibited by dimer-sulfides. All screened compounds exerted concentration-dependent cytotoxic activity against leukemia K562 cells. The compounds which exerted the most pronounced cytotoxic action exhibited notably higher cytotoxic activities against K562, HeLa and MDA-MB-453 cells in comparison to resting and PHA-stimulated PBMC, pointing to a significant selectivity in their antitumor actions. Examination of the mechanisms of cytotoxicity on leukemia K562 cells revealed pro-apoptotic action of each of the investigated compounds applied at concentrations 2IC50. The most prominent pro-apoptotic action was exhibited by dimer-sulfide of cholest-4-en-3-one. Furthermore, almost all of the tested compounds at IC50 concentrations induced G1 phase cell cycle arrest in K562 cells. Antimicrobial activity against Gram-positive, Gram-negative bacteria and fungal cells, and toxicity to brine shrimp Artemia salina, were evaluated. There was no antibacterial activity. The best antifungal activity was exhibited against Saccharomyces cerevisiae by dimers linked with trithiolane ring, indicating a selective activity of investigated compounds.


Mail-order medicine; An analysis of the Sears Roebuck Foundation's Community Medical Assistance Program.
Of the 625 rural communities that originally applied to the Sears Roebuck Foundation's Community Medical Assistance Program, 253 were accepted. One hundred sixty-three went on to build clinics, and 132 had physicians at the time of follow-up. From the standpoint of physician coverage of the community once a clinic had been built, the rate of physician availability was 78 percent, The communities' success in recruiting and retaining physicians was compared against 13 variables; none was significantly related to the recruitment, and only two--numbers of service clubs and distance to nearest physician--were related to retention.


The efficacy of telavancin in comparison with linezolid on endotracheal tube biofilm in pigs with methicillin-resistant Staphylococcus aureus pneumonia.
The effect of systemic treatment of ventilator-associated pneumonia (VAP) with telavancin, a semisynthetic lipoglycopeptide with good penetration in vitro biofilms, has not been tested in vivo during mechanical ventilation. This study examined the efficacy of telavancin compared with linezolid against endotracheal tube (ETT) biofilms in a porcine model of methicillin- resistant Staphylococcus aureus (MRSA) VAP. VAP was induced in 18 pigs by instilling 107 colony-forming units (CFU/mL) of a MRSA strain susceptible to telavancin and linezolid into each pulmonary lobe. Randomization into three groups was done at pneumonia diagnosis: control (IV glucose 0.5% solution q24); linezolid (10 mg/kg q12) and telavancin groups (22.5 mg/kg q24). After 72h of MV, data regarding bronchoalveolar lavage (BAL), tracheal aspirate (TA), ETT MRSA biofilm load and thickness measured by scanning electron microscopy were obtained. All 18 pigs completed the study. MRSA was isolated in 100% of ETTs from the control and linezolid groups, and in 67% from the telavancin group. Telavancin treatment presented lower MRSA load compared to the control and linezolid treatments (telavancin median [interquartile range (IQR)] =1.94 [0.00-5.45], linezolid 3.99 [3.22-4.68] and control 4.93 [4.41-5.15], p=0.236). Telavancin treatment also resulted in the lowest biofilm thickness according to the SEM (4.04 [2.09-6.00], p<0.001). We found a positive correlation between ETT and BAL load (rho=0.511, p=0.045). In our VAP model, systemic telavancin treatment reduced ETT MRSA occurrence, load, and biofilm thickness. Our findings may have a bearing on ICU patients' clinical outcomes.


RNA processing in human mitochondria.
Mammalian mitochondrial DNA is transcribed as precursor polycistronic transcripts containing 13 mRNAs, 2 rRNAs, punctuated by 22 tRNAs. The mechanisms involved in the excision of mitochondrial tRNAs from these polycistronic transcripts have remained largely unknown. We have investigated the roles of ELAC2, mitochondrial RNase P proteins 1 and 3, and pentatricopeptide repeat domain protein 1 in the processing of mitochondrial polycistronic transcripts. We used a deep sequencing approach to characterize the 5' and 3' ends of processed mitochondrial transcripts and provide a detailed map of mitochondrial tRNA processing sites affected by these proteins. We show that MRPP1 and MRPP3 process the 5' ends of tRNAs and the 5' unconventional, non tRNA containing site of the CO1 transcript. By contrast, we find that ELAC2 and PTCD1 affect the 3' end processing of tRNAs. Finally, we found that MRPP1 is essential for transcript processing, RNA modification, translation and mitochondrial respiration.


Strategy and the Internet.
Many of the pioneers of Internet business, both dot-coms and established companies, have competed in ways that violate nearly every precept of good strategy. Rather than focus on profits, they have chased customers indiscriminately through discounting, channel incentives, and advertising. Rather than concentrate on delivering value that earns an attractive price from customers, they have pursued indirect revenues such as advertising and click-through fees. Rather than make trade-offs, they have rushed to offer every conceivable product or service. It did not have to be this way--and it does not have to be in the future. When it comes to reinforcing a distinctive strategy, Michael Porter argues, the Internet provides a better technological platform than previous generations of IT. Gaining competitive advantage does not require a radically new approach to business; it requires building on the proven principles of effective strategy. Porter argues that, contrary to recent thought, the Internet is not disruptive to most existing industries and established companies. It rarely nullifies important sources of competitive advantage in an industry; it often makes them even more valuable. And as all companies embrace Internet technology, the Internet itself will be neutralized as a source of advantage. Robust competitive advantages will arise instead from traditional strengths such as unique products, proprietary content, and distinctive physical activities. Internet technology may be able to fortify those advantages, but it is unlikely to supplant them. Porter debunks such Internet myths as first-mover advantage, the power of virtual companies, and the multiplying rewards of network effects. He disentangles the distorted signals from the marketplace, explains why the Internet complements rather than cannibalizes existing ways of doing business, and outlines strategic imperatives for dot-coms and traditional companies.


Pressure effect on the nonradiative process of thioflavin-T.
Time-resolved emission techniques were employed to study the nonradiative process of thioflavin-T (ThT) in 1-propanol, 1-butanol, and 1-pentanol as a function of the hydrostatic pressure. Elevated hydrostatic pressure increases the alcohol viscosity, which in turn strongly influences the nonradiative rate of ThT. A diamond-anvil cell was used to increase the pressure up to 2.4 GPa. We found that the nonradiative rate constant, k(nr), decreases with pressure. We further found a remarkable linear correlation between a decrease in k(nr) (increase in the nonradiative lifetime, τ(nr)) and an increase in the solvent viscosity. The viscosity was varied by a factor of 1000 and k(nr) was measured at high pressures, at which the nonradiative rate constant of the molecules decreased from (7 ps)(-1) to (13 ns)(-1), (13 ps)(-1) to (17 ns)(-1) and (17 ps)(-1) to (15 ns)(-1) for 1-propanol, 1-butanol, and 1-pentanol, respectively. The viscosity-dependence of k(nr) is explained by the excited-state rotation rate of the two-ring systems, with respect to each other.


Nrf2 protects human bladder urothelial cells from arsenite and monomethylarsonous acid toxicity.
Arsenic is widely spread in our living environment and imposes a big challenge on human health worldwide. Arsenic damages biological systems through multiple mechanisms including the generation of reactive oxygen species. The transcription factor Nrf2 regulates the cellular antioxidant response that protects cells from various insults. In this study, the protective role of Nrf2 in arsenic toxicity was investigated in a human bladder urothelial cell line, UROtsa. Using a UROtsa cell line stably infected with Nrf2-siRNA, we clearly demonstrate that compromised Nrf2 expression sensitized the cells to As(III)- and MMA(III)-induced toxicity. On the other hand, the activation of the Nrf2 pathway by tert-butylhydroquinone (tBHQ) and sulforaphane (SF), the known Nrf2-inducers, rendered UROtsa cells more resistant to As(III) and MMA(III). Furthermore, the wild-type mouse embryo fibroblast (WT-MEF) cells were protected from As(III)- and MMA(III)-induced toxicity following Nrf2 activation by tBHQ or SF, whereas neither tBHQ nor SF conferred protection in the Nrf2(-/-)MEF cells, demonstrating that tBHQ- or SF-mediated protection against As(III)- and MMA(III)-induced toxicity depends on Nrf2 activation. These results, obtained by both loss of function and gain of function analyses, clearly demonstrate the protective role of Nrf2 in arsenic-induced toxicity. The current work lays the groundwork for using Nrf2 activators for therapeutic and dietary interventions against adverse effects of arsenic.


Stabilization of composted organic matter after application to a humus-free sandy mining soil.
The use of mining substrates for recultivation purposes is limited due to their low organic matter (OM) contents. In a 1-yr laboratory experiment we evaluated the stabilization of biowaste compost added to a humus-free sandy mining soil to examine the suitability of compost amendment for the formation of stable soil organic matter (SOM). The stabilization process was characterized by measuring enrichment of OM and nitrogen in particle size fractions obtained after dispersion with different amounts of energy (ultrasonication and shaking in water), carbon mineralization, and amount of dissolved organic carbon (DOC). During the experiment, 17.1% of the organic carbon (OC) was mineralized. Organic carbon enrichment in the < 20-micron particle size fraction at the beginning of the experiment was in the range of natural soils with similar texture. Within 12 mo, a distinct OC redistribution from coarse into fine fractions was found with both dispersion methods. The accumulation of OC was more pronounced for the size separates obtained by ultrasonication, where the carbon distribution between 0.45- to 20-micron particle size fractions increased from 30% at the beginning to 71% at the end of the experiment. Dissolved organic carbon contents ranged between 50 and 68 g kg-1 OC and decreased during the incubation. In conclusion, the exponential decrease of carbon mineralization and the OC enrichment in the fine particle size fractions both indicated a distinct OM stabilization in the mining soil.


Systematic review and meta-analysis of clinical significance of autoantibodies for idiopathic pulmonary fibrosis.
To clarify clinical significance of the sole presence of autoantibodies for idiopathic pulmonary fibrosis (IPF) without any other symptoms or signs suggestive of autoimmune disease. Systematic review and meta-analysis DATA SOURCES: Medline, EMBASE, Science Citation Index Expanded and Google Scholar were searched from 1 January 2002 through 12 February 2019. Primary studies addressing all-cause mortality and the development of a defined autoimmune disease for IPF with autoantibodies were included for the review. Two reviewers extracted relevant data and assessed risk of bias independently. Meta-analysis was conducted using a random-effects model if three or more studies reported the same outcome for a certain autoantibody. The quality of evidence was assessed by the Grades of Recommendation, Assessment, Development and Evaluation system. Out of 4603 records retrieved nine studies were included in this review. All studies contained some risk of bias. Based on pooled data myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA) was significantly associated with microscopic polyangiitis incidence with risk ratio (RR) of 20.2 (95% CI: 7.22 to 56.4) and antinuclear antibody (ANA) was also significantly associated with the development of connective tissue diseases with RR of 7.11 (p=0.001) (10 cases in 157 patients with ANA) in one study. However, there was no significant association of autoantibodies with all-cause mortality aside from MPO-ANCA and proteinase 3-ANCA in one study each. MPO-ANCA was not demonstrated to be associated with all-cause mortality by meta-analysis. The quality of evidence was deemed as either low or very low. The presence of autoantibodies such as MPO-ANCA and ANA was demonstrated to be associated with the development of some autoimmune diseases for patients with IPF although there was no difference of all-cause mortality. However, the results should be interpreted with caution due to low evidence level. CRD42017077336.


Enteroviruses and type 1 diabetes.
The development of type 1 diabetes mellitus (T1DM) has been linked to exposure to environmental triggers, with Enteroviruses (EV) historically considered the prime suspects. Early serological studies suggested a link between EV infections and the development of T1DM and, though controversial, have been bolstered by more recent studies using more sensitive techniques such as direct detection of the EV genome by RT-PCR in peripheral blood. In this review, we consider the weight of evidence that EV can be considered a candidate trigger of T1DM, using three major criteria: (1) is EV infection associated with clinical T1DM, (2) can EV trigger the development of autoimmunity and (3) what would explain the putative association?


Neonatal cholestasis in children with Alpha-1-AT deficiency is a risk for earlier severe liver disease with male predominance.
Our objective was to better understand the natural history and disease modifiers of Alpha-1-antitrypsin deficiency (AATD), a common genetic liver disease causing hepatitis and cirrhosis in adults and children. The clinical course is highly variable. Some infants present with neonatal cholestasis, which can resolve spontaneously or progress to cirrhosis; others are well in infancy, only to develop portal hypertension later in childhood. The Childhood Liver Disease Research Network has been enrolling AATD participants into longitudinal, observational studies at North American tertiary centers since 2004. We examined the clinical courses of 2 subgroups of participants from the several hundred enrolled; first, those presenting with neonatal cholestasis captured by a unique study, enrolled because of neonatal cholestasis but before specific diagnosis, then followed longitudinally (n=46); second, separately, all participants who progressed to liver transplant (n=119). We found male predominance for neonatal cholestasis in AATD (65% male, p=0.04), an association of neonatal gamma-glutamyl transpeptidase elevation to more severe disease, and a higher rate of neonatal cholestasis progression to portal hypertension than previously reported (41%) occurring at median age of 5 months. Participants with and without preceding neonatal cholestasis were at risk of progression to transplant. Participants who progressed to liver transplant following neonatal cholestasis were significantly younger at transplant than those without neonatal cholestasis (4.1 vs. 7.8 years, p=0.04, overall range 0.3-17 years). Neonatal cholestasis had a negative impact on growth parameters. Coagulopathy and varices were common before transplant, but gastrointestinal bleeding was not. Patients with AATD and neonatal cholestasis are at risk of early progression to severe liver disease, but the risk of severe disease extends throughout childhood. Careful attention to nutrition and growth is needed.


Role of renal nerves in compensatory adaptation to chronic reductions in sodium intake.
The aim of this study was to investigate the importance of the renal nerves in adaptation to chronic reductions in sodium intake. Conscious dogs with unilateral (n = 7) or bilateral (n = 4) renal denervation were studied. In dogs studied before and after bilateral denervation, there were no differences in urine volume (UO), Na excretion (UNaV), or fractional reabsorption of Li (FR Li, an index of proximal tubular Na reabsorption) between innervated and denervated kidneys on either normal (80 meq/day) or low Na intake (5 meq/day, 15 days). Plasma renin activity (PRA) was attenuated following denervation on both normal (0.39 +/- 0.06 vs. 0.18 +/- 0.01 ng angiotensin I X ml-1 X h-1) and low Na intake (1.00 +/- 0.06 vs. 0.59 +/- 0.01). In unilaterally denervated dogs the left kidney was denervated and the bladder was split, allowing continuous urine collection from separate innervated and denervated kidneys in the same dog. There was no difference in UO between innervated and denervated kidneys on normal (80 meq/day) or low (7 meq/day, 9 days) Na intake. UNaV averaged 33.6 +/- 1.3 and 37.6 +/- 2.1 meq/day in innervated and denervated kidneys, respectively, on normal Na intake and 3.5 +/- 0.5 and 4.0 +/- 0.4 meq/day in innervated and denervated kidneys on low Na intake. FR Li was not different in denervated compared with innervated kidneys during normal or low sodium intake. Norepinephrine content was reduced by 99 +/- 1% in denervated kidneys.(ABSTRACT TRUNCATED AT 250 WORDS)


Isosporiasis in Venezuelan adults infected with human immunodeficiency virus: clinical characterization.
A cross-sectional study was undertaken to determine the prevalence of isosporiasis and its clinical and laboratory pattern in Venezuelan patients infected with human immunodeficiency virus (HIV) (n = 397). At enrollment, they underwent a thorough clinical history and physical examination, and provided stool specimens for the identification of Isospora belli and other parasites. Isospora belli was identified in 56 subjects (14%) and diarrhea, either acute or chronic, was present in 98% of these cases (P < 0.001). Eosinophilia was strongly associated with isosporiasis (P = 0.01). It was also found that the presence of eosinophilia was more common in I. belli-infected patients without weight loss (P < 0.001). Twenty-six (81.25%) subjects with I. belli infection had CD4+ cell counts < 200 cells/mm3 (P = 0.03). In addition, the data and its description shows the association to be < 100 cells/mm3. This infection seems to be seasonal because the recovery of oocysts occurred mainly in months with significant rainfall. In fact, isosporiasis should be suspected in HIV-infected patients from tropical countries with diarrhea, weight loss, eosinophilia, and low CD4+ cell counts.


Molecular evolution of Na+ channels in teleost fishes.
Voltage-dependent sodium channels are critical for electrical excitability. Invertebrates possess a single sodium channel gene; two rounds of genome duplication early in vertebrates increased the number to four. Since the teleost-tetrapod split, independent gene duplications in each lineage have further increased the number of sodium channel genes to 10 in tetrapods and 8 in teleosts. Here we review how the occurrence of multiple sodium channel paralogs has influenced the evolutionary history of three groups of fishes: pufferfish, gymnotiform and mormyriform electric fish. Pufferfish (tetraodontidae) produce a neurotoxin, tetrodotoxin, that binds to and blocks the pore of sodium channels. Pufferfish evolved resistance to their own toxins by amino acid substitutions in the pore of their sodium channels. These substitutions had to occur in parallel across multiple paralogs for organismal resistance to evolve. Gymnotiform and mormyriform fishes independently evolved electric organs to generate electricity for communication and object localization. Two sodium channel genes are expressed in muscle in most fishes. In both groups of weakly electric fishes, one gene lost its expression in muscle and became compartmentalized in the evolutionary novel electric organ, which is a muscle derivative. This gene then evolved at elevated rates, whereas the gene that is still expressed in muscle does not show elevated rates of evolution. In the electric organ-expressing gene, amino acid substitutions occur in parts of the channel involved in determining how long the channel will be open or closed. The enhanced rate of sequence evolution of this gene likely underlies the species-level variations in the electric signal.


Use of a serotype-specific urine immunoassay to determine the course of a hospital outbreak of Streptococcus pneumoniae complicated by influenza A.
An outbreak of Streptococcus pneumoniae (pneumococcal) infection complicated by concomitant influenza A on an elderly care ward was detected. Thirteen patients with hospital-acquired respiratory infections were investigated during the course of the outbreak investigation. Six had a positive BinaxNOW S. pneumoniae urinary antigen test and two patients had culture-confirmed pneumococcal bacteraemia and a positive urine antigen test. Five patients gave positive influenza A PCR results of which two were also positive for S. pneumoniae antigen. The concurrence of influenza and pneumococcal infections made tracking the course of the infection difficult. This case study shows how the use of a sensitive, S. pneumoniae serotype-specific urine antigen assay, in the absence of cultured isolates, helped determine whether patients were infected with the same pneumococcal serotype. This was particularly useful when additional respiratory symptoms were seen following the administration of chemoprophylaxis.


Cloning, expression, and gene structure of a G protein-coupled glutamate receptor from rat brain.
A complementary DNA encoding a G protein-coupled glutamate receptor from rat brain, GluGR, was cloned by functional expression in Xenopus oocytes. The complementary DNA encodes a protein of 1199 amino acids containing a seven-transmembrane motif, flanked by large amino- and carboxyl-terminal domains. This receptor lacks any amino acid sequence similarity with other G protein-coupled receptors, suggesting that it may be a member of a new subfamily. The presence of two introns flanking the central core suggests that GluGR may have evolved by exon shuffling. Expressed in oocytes, GluGR is activated by quisqualate greater than glutamate greater than ibotenate greater than trans-1-aminocyclopentyl-1,3-dicarboxylate, and it is inhibited by 2-amino-3-phosphonopropionate. Activation is blocked by Bordella pertussis toxin. These properties are typical of some metabotropic glutamate receptors.


Microbiomic subprofiles and MDR1 promoter methylation in head and neck squamous cell carcinoma.
Clinical observations and epidemiologic studies suggest that the incidence of head and neck squamous cell carcinoma (HNSCC) correlates with dental hygiene, implying a role for bacteria-induced inflammation in its pathogenesis. Here we begin to explore the pilot hypothesis that specific microbial populations may contribute to HNSCC pathogenesis via epigenetic modifications in inflammatory- and HNSCC-associated genes. Microbiomic profiling by 16S rRNA sequencing of matched tumor and adjacent normal tissue specimens in 42 individuals with HNSCC demonstrate a significant association of specific bacterial subpopulations with HNSCC over normal tissue (P < 0.01). Furthermore, microbial populations can separate tumors by tobacco status (P < 0.008), but not by alcohol status (P = 0.41). If our subhypothesis regarding a mechanistic link from microorganism to carcinogenesis via inflammation and consequent aberrant DNA methylation is correct, then we should see hypermethylation of relevant genes associate with specific microbiomic profiles. Methylation analysis in four genes (MDR1, IL8, RARB, TGFBR2) previously linked to HNSCC or inflammation shows significantly increased methylation in tumor samples compared with normal oral mucosa. Of these, MDR1 promoter methylation associates with specific microbiomic profiles in tumor over normal mucosa. Additionally, we report that MDR1 methylation correlates with regional nodal metastases in the context of two specific bacterial subpopulations, Enterobacteriaceae and Tenericutes (P < 0.001 for each). These associations may lead to a different, and potentially more comprehensive, perspective on the pathogenesis of HNSCC, and support further exploration of mechanistic linkage and, if so, novel therapeutic strategies such as demethylating agents and probiotic adjuncts, particularly for patients with advanced or refractory disease.


Glycolipid-anchored form of CD4 increases intercellular adhesion but is unable to enhance T cell activation.
CD4 functions to enhance T cell activation by increasing intercellular adhesion and/or by transduction of an intracellular signal. To study the role of human CD4 in T cell activation we have used a murine T cell hybridoma, By 155.16, which produces IL-2 when stimulated by HLA-DR-bearing cells. Previously, we have shown that expression of human CD4 by this hybridoma enhances its ability to produce IL-2 in response to HLA-DR-bearing cells. Furthermore, deletion of the majority of the cytoplasmic domain renders CD4 less efficient at enhancing IL-2 production. We describe studies of a glycolipid-anchored mutant of the CD4 molecule, CD4PI. This mutant is composed of the entire extracellular domain of CD4 anchored to the outlet leaflet of the membrane via a covalent bond to glycosylphosphatidylinositol and, therefore, has no transmembrane or cytoplasmic domains. When expressed in By155.16, CD4PI shows no defect in its ability to increase intercellular adhesion but is unable to augment IL-2 production. These results clearly demonstrate that CD4 enhances T cell activation by mechanisms other than increasing intercellular adhesion.


Comparison of percutaneous endoscopic interlaminar discectomy and conventional open lumbar discectomy for L4/5 and L5/S1 double-segmental lumbar disk herniation.
Although spinal endoscopic techniques have shown great advantages in the treatment of single-segment lumbar disk herniation (LDH), the therapeutic advantages for double-segment LDH are controversial. To compare the outcomes of percutaneous endoscopic interlaminar discectomy (PEID) versus conventional open lumbar discectomy (COLD) for the treatment of L4/5 and L5/S1 double-segmental LDH. From January 2016 to September 2021, we included 50 patients with double-segmental LDH who underwent PEID (n = 25) or COLD (n = 25). The clinical outcomes between the two groups were evaluated using the visual analog scale (VAS), the Oswestry disability index (ODI), and the modified MacNab criteria. Moreover, the incision length, operation time, intraoperative fluoroscopy time, postoperative bedtime, hospital stays, and complications were also recorded and compared after surgery. In both groups, the VAS and ODI scores at different timepoints postoperatively were significantly improved compared with those preoperatively (P < 0.05) According to the modified MacNab criteria, the excellent or good outcome rate was 92% in the PEID group and 88% in the COLD group. The PEID group had shorter incision length, postoperative bedtime, and hospital stays than the COLD group. However, the operation time was shorter and intraoperative fluoroscopy time was fewer in the COLD group. In addition, there was no significant difference between the two groups in terms of surgical complications during the postoperative follow-up period. Both PEID and COLD have good efficacy and high safety for management of L4/5 and L5/S1 double-segmental LDH. Compared with the COLD group, the PEID group had more operative time as well as more intraoperative fluoroscopy, but it had a more minimally invasive surgical incision as well as faster postoperative recovery.


Massive Muscular Infection by a Sarcocystis Species in a South American Rattlesnake (Crotalus durissus terrificus).
Massive numbers of sarcocysts of a previously undescribed species of Sarcocystis were observed in the skeletal muscles throughout the body of an adult, female South American rattlesnake (Crotalus durissus terrificus). Examination of tissue sections by light microscopy demonstrated that sarcocysts were present in 20 to 40% of muscle fibers from 5 sampled locations. Sarcocysts were not present in cardiac muscle, smooth muscle, or other organs. Sarcocysts were 0.05-0.15 mm wide, had variable length depending on the viewed orientation and size of the muscle fiber, and had a sarcocyst wall less than 1-μm thick. Sarcocysts were subdivided by septa and had central degeneration in older sarcocysts. Host induced secondary encapsulation or an inflammatory response was not present. By transmission electron microscopy (TEM), the sarcocyst wall was Type I, with a parasitophorous membrane of approximately 100 nanometers in width arranged in an undulating pattern and intermittently folded inward in a branching pattern. The sarcocysts contained metrocytes in different stages of development and mature bradyzoites. The nucleic acid sequence from a section of the 18S small subunit rRNA gene was most closely related to S. mucosa that uses marsupials as intermediate hosts and has an unknown definitive host. This is apparently the third report of muscular Sarcocystis infection in snakes and is the first to describe the ultrastructure of the sarcocysts and use sequencing methods to aid in identification.


[The current state and prospects of the development of forensic medical expertise of the blast injury].
The analysis of the special literature and expert practice during the recent years demonstrated the scientific, practical, and social importance of the expertise of the blast injuries in the time of piece. It is emphasized that the main tasks of expert examination of blast injuries include further studies on the mechanisms of their development, simulation of various blast injuries, the development of criteria for the determination of distance from the site of explosion based on the severity of the injury, investigations into effectiveness of the protective barriers for the human body during explosions, and the creation of methods for testing means of individual body armoured protection. The provisions of a program for current and prospective forensic medical investigations into blast injuries have been formulated.


Quantitative expression profile of androgen-regulated genes in prostate cancer cells and identification of prostate-specific genes.
Quantitative expression profile of androgen-regulated genes (ARGs) was evaluated in the hormone-responsive prostate cancer cell line LNCaP by serial analysis of gene expression (SAGE). A total of 83,489 SAGE tags representing 23,448 known genes or expressed sequence tags (ESTs) and 1,655 potentially novel sequences have unraveled the transcriptome of LNCaP cells, the most common cell line used in prostate cancer research. Comparison of transcripts between control and R1881-treated LNCaP cells revealed the induction of 136 genes and repression of 215 genes in response to androgen (p < 0.05). Strikingly, a high fraction ( approximately 90%) of ARGs identified in our study has not been described as ARGs previously. A number of prostate-specific transcription factors were among the ARGs identified here. Classification of the ARGs on the basis of biochemical functions revealed that a great majority of ARGs identified in our experimental system appear to be involved in regulation of transcription, splicing, ribosomal biogenesis, mitogenesis, bioenergetics and redox processes. One of the novel aspects of androgen signaling included androgen regulation of genes involved in DNA repair/recombination process. By comparing our LNCaP-C and LNCaP-T SAGE libraries with SAGE tag libraries available at the NCBI-SAGE website, we have identified >200 potential prostate specific/abundant transcripts. The discovery of new prostate-specific genes and ARGs provides a unique opportunity to determine the role of these genes in prostate cell growth, differentiation and tumorigenesis.


The use of adenoviral vectors and ex vivo transduced neurotransplants: towards promotion of neuroregeneration.
Regeneration of injured axons following injury depends on a delicate balance between growth-promoting and growth-inhibiting factors. Overexpression of neurotrophin genes seems a promising strategy to promote regeneration. Trophic genes can be overexpressed at the site of injury at the axonal stumps, or at the perikaryal level of the injured neuron. Transduction of the neural cells can be achieved by applying adenoviral vectors, either directly in vivo or-in the case of neurotransplantation as an ex vivo approach. In both cases it would create a more permissive environment for axonal growth and therefore in functional regeneration. In this article, the feasibility of the use of adenoviral vectors in several neuroregeneration models--in particularly in spinal cord lesion models and the biological clock transplantation model--is illustrated. The results show that the adenoviral vectors can be a powerful tool to study the effects of overexpression of genes in an in vivo paradigm of nerve regeneration or nerve outgrowth. The potential use of adenoviral vectors and ex vivo transduced neurotransplants is discussed.


Comparative analysis of encephalization in mammals reveals relaxed constraints on anthropoid primate and cetacean brain scaling.
There is a well-established allometric relationship between brain and body mass in mammals. Deviation of relatively increased brain size from this pattern appears to coincide with enhanced cognitive abilities. To examine whether there is a phylogenetic structure to such episodes of changes in encephalization across mammals, we used phylogenetic techniques to analyse brain mass, body mass and encephalization quotient (EQ) among 630 extant mammalian species. Among all mammals, anthropoid primates and odontocete cetaceans have significantly greater variance in EQ, suggesting that evolutionary constraints that result in a strict correlation between brain and body mass have independently become relaxed. Moreover, ancestral state reconstructions of absolute brain mass, body mass and EQ revealed patterns of increase and decrease in EQ within anthropoid primates and cetaceans. We propose both neutral drift and selective factors may have played a role in the evolution of brain-body allometry.


[Application of a technique to establish criteria for uterine length measurement].
The uterine length measurement is a routine practice in the prenatal care of pregnant women. It has been attributed a sensibility of 86% to detect fetal grown restriction. The technique is easy to perform and reproducible between observers, although we have found variation between intra- and inter-observers measurements. To evaluate the useful of a maneuver to improve the reliability of the uterine length measurement in a group of obstetrics residents. The measurement was performed on pregnant women. The results were analyzed by intraclass correlation coefficient to evaluate the concordance between observers based in a variance analysis model (ANOVA) of repeated measurements. There were not differences between groups about demographic variables. All residents improve the reliability in their measurements. The intra-class correlation coefficient was 0.77 (confidence interval 0.63-0.88) before the maneuver, and after this one the intra-class correlation coefficient improved to 0.96 (confidence interval 0.92-0.98). The uterine measurement following detailed indications improve the interobserver concordance. We think it is advisable to evaluate the use of this maneuver in a routine way in our institute and in other places that bring prenatal care.


Chondroitin sulfate proteoglycan expression and binding of Plasmodium falciparum-infected erythrocytes in the human placenta during pregnancy.
A characteristic feature of malaria during pregnancy is the sequestration of Plasmodium falciparum-infected red blood cells (IRBCs) in the intervillous spaces of the placenta. We have recently shown that unusually low-sulfated chondroitin sulfate proteoglycans (CSPGs) present in the intervillous spaces mediate the adherence of IRBCs in the placenta. In areas of endemicity, the prevalence of P. falciparum infection in pregnant women peaks during weeks 13 to 20 and then gradually declines, implying that the placental CSPGs are available for IRBC adhesion early during the pregnancy. However, there is no information on the expression and composition of CSPGs during pregnancy. In this study, the expression pattern of CSPGs during the course of pregnancy was investigated. The CSPGs were purified from placentas of various gestational ages, characterized, and tested for the ability to bind IRBCs. The data demonstrate that the CSPGs are present in the intervillous spaces throughout the second and third trimesters. The levels of CSPGs expressed per unit tissue weight were similar in placentas of various gestational ages. However, the structures of the intervillous-space CSPGs changed considerably during the course of pregnancy. In particular, the molecular weight was decreased, with an accompanying gradual increase in the CSPG size polydispersity, from 16 weeks until 38 weeks. The sulfate content was increased considerably after 24 weeks. Despite these structural changes, the CSPGs of placentas of various gestational ages efficiently supported the binding of IRBCs. These results demonstrate that CSPGs can mediate the sequestration of IRBCs in the intervillous spaces of the placenta during the entire second and third trimesters and possibly during the later part of the first trimester as well.


A framework to simplify paediatric syncope diagnosis.
This paper aims to improve the diagnosis of syncope and transient loss of consciousness (TLOC) in children. Diagnostic problems stem, first, from some causes spanning various disciplines, e.g. cardiology, neurology and psychiatry, while the most common cause, vasovagal syncope, is not embraced by any specialty. Second, clinical variability is huge with overlapping signs and symptoms. Third, the approach to TLOC/syncope of the European Society of Cardiology (ESC) is underused in childcare. We explain the ESC guidelines using an additional paediatric literature review. Classification of TLOC and syncope is hierarchic and based on history taking. Loss of consciousness (LOC) is defined using three features: abnormal motor control including falling, reduced responsiveness and amnesia. Adding a < 5 min duration and spontaneous recovery defines TLOC. TLOC simplifies diagnosis by excluding long LOC (e.g. some trauma, intoxications and hypoglycaemia) and focussing on syncope, tonic-clonic seizures and functional TLOC. Syncope, i.e. TLOC due to cerebral hypoperfusion, is divided into reflex syncope (mostly vasovagal), orthostatic hypotension (mostly initial orthostatic hypotension in adolescents) and cardiac syncope (arrhythmias and structural cardiac disorders). The initial investigation comprises history taking, physical examination and ECG; the value of orthostatic blood pressure measurement is unproven in children but probably low. When this fails to yield a diagnosis, cardiac risk factors are assessed; important clues are supine syncope, syncope during exercise, early death in relatives and ECG abnormalities. Conclusions: In adults, the application of the ESC guidelines reduced the number of absent diagnoses and costs; we hope this also holds for children. What is Known: • Syncope and its mimics are very common in childhood, as they are at other ages. • Syncope and its mimics provide considerable diagnostic challenges. What is New: • Application of the hierarchic framework of transient loss of consciousness (TLOC) simplifies diagnosis. • The framework stresses history-taking to diagnose common conditions while keeping an eye on cardiac danger signs.


Baicalin provides neuroprotection in traumatic brain injury mice model through Akt/Nrf2 pathway.
The neuroprotective effects of Baicalin have been confirmed in several central nervous system (CNS) diseases. However, its possible effect on traumatic brain injury (TBI) model is still not clear. The present study is aimed to investigate the role and the underling mechanisms of 7-D-glucuronic acid-5,6-dihydroxyflavone (Baicalin) on TBI model. The weight-drop model of TBI in Institute of Cancer Research mice was treated with Baicalin intraperitoneally at 30 minutes after TBI. LY294002 (LY) (a commonly used PI3K/Akt pathway inhibitor) was injected into the left ventricle at 30 minutes before TBI. All mice were euthanized at 24 hours after TBI to collect the brain tissue for a series of tests except for neurological function, which was measured at 2 hours and 1 and 3 days post-TBI. Baicalin administration significantly improved neurobehavioral function, alleviated brain edema, and reduced apoptosis-positive cells by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay accompanied with the upregulation of B-cell lymphoma 2 (Bcl-2) and downregulation of Bcl-2-associated X protein (Bax) and cleaved-caspase 3 by Western blot. Besides, TBI-induced oxidant stress status was also restored in the Baicalin group by measuring malondialdehyde (MDA) content, glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels in the injured brain cortex. Furthermore, translocation of Nrf2 to the nucleus was dramatically enhanced by Baicalin verified by immunofluorescence and Western blot analyses. Accordingly, its downstream antioxidative enzymes nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase 1 (NQO-1) and heme oxygenase 1 (HO-1) were also activated by Baicalin confirmed by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot. However, cotreatment with Baicalin and LY could partly abolish Baicalin-induced activation of Nrf2 and its neuroprotective effects in TBI. This study demonstrates that Baicalin provides a neuroprotective effect in TBI mice model via activating the Akt/Nrf2 pathway.


Techniques of selective cannulation and sphincterotomy.
Selective access into the desired duct followed by incision of the sphincter, i. e. sphincterotomy, forms the cornerstone of any endoscopic intervention within the pancreaticobiliary system. The apprehensive beginner's performance and hesitance is aggravated by ignorance of ampullary anatomy and he considers selective cannulation to be the greatest hurdle. An understanding of ampullary morphology and its variations is vital in achieving selective cannulation. Technological advances have assisted in the form of development of better accessories, progressing from "immovable" catheters to movable cannulae and to single-, double-, and even triple-lumen sphincterotomes. Orientation along the long axis of the bile duct ensures access and avoids inadvertent and hazardous manipulation of the pancreatitic duct. Using guide wires, especially the 'angulated-tip' glide wire improves cannulation successs rates considerably. Precut accessotomy complements wire-guided selective cannulation, and can be used analogously to a controlled surgical incision to facilitate cannulation of the desired system after deroofing the papilla layer by layer. Published data have validated its role, demonstrating high efficacy and minimal complications when it is properly performed. Biliary sphincterotomy, using the right mode of blended current in the 11-12 o'clock direction and with the tip of the sphincterotome wire, provides a clean and bloodless splitting open of the sphincter of Oddi. Pancreatic precut, over-the-stent papillotomy and sphincterotomy over a guide wire have all been proven to be safe and effective measures, in large groups of patients. In special situations, such as where there are impacted stones or ampullary lesions, needle-knife infundibulotomy achieves reliable access. Techniques such as saline infiltration into the papilla and subtle body movements to re-position the scope enable biliary cannulation in difficult situations. Alterations in anatomy, for instance post Billroth II gastrectomy, no longer discourage the endoscopist from attempting intervention. Application of knowledge of reverse anatomy, specially designed instruments, and adherence to the proper technique improves success in these patients. Our experience of 9000 sphincterotomies over the past 12 years with minimal morbidity stands proof to the principles and techniques highlighted in this monograph. We recommend these to all aspiring endoscopists, with the assurance of improved technical success when they are implemented.


A municipal health center's co-constructed therapeutic patient education program for patients with single or multiple morbidities
The accessibility of Therapeutic Patient Education (TPE) is essential to reduce social inequalities and to meet the challenges posed by the increase in chronic diseases. In France, the distribution of TPE throughout the territory is heterogeneous and patients still lack sufficient access. With these perspectives in mind, a municipal health center has developed an innovative TPE device by involving concerned patients from the outset. The objectives of this article are to present the methodology implemented to co-construct a TPE program for patients with one or more prevalent diseases in the territory, as well as the results of the co-construction. Creation of a project team with various actors: patients, health professionals, administrators, researchers. Organization of synchronous and asynchronous times. Four meetings and numerous exchanges made it possible to: define the common values and operating rules of the project team, reflect on the coordination of the care pathway, identify the specific and shared needs of patients affected by type 2 diabetes, arterial hypertension and/or obesity, agree on the principles of the device. The advantages of this device were discussed: flexible organization with numerous links with local actors, development of partnerships, expansion of the population concerned. Questions and difficulties were encountered, such as the coordination of actors and the representation of partner patients for the targeted diseases. This device will be tested and evaluated, mainly with an objective of improving it.


Comparison of ceftriaxone and ampicillin plus chloramphenicol for the therapy of acute bacterial meningitis.
Ceftriaxone, a new third-generation cephalosporin, appears to be promising for the therapy of acute bacterial meningitis. The 90% MBCs of ceftriaxone against 54 recent cerebrospinal fluid isolates of Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae were less than or equal to 0.06 to 0.25 micrograms/ml. We examined the efficacy and safety of ceftriaxone therapy of meningitis in Bahia, Brazil. The study was conducted in two phases; in phase A, ceftriaxone was coadministered with ampicillin. The mean cerebrospinal fluid concentrations of ceftriaxone 24 h after an intravenous dose of 80 mg/kg were 4.2 and 2.3 micrograms/ml on days 4 to 6 and 10 to 12 of therapy, respectively. These concentrations were 8- to more than 100-fold greater than the 90% MBCs against the relevant pathogens. In phase B, ceftriaxone (administered once daily at a dose of 80 mg/kg after an initial dose of 100 mg/kg) was compared with conventional dosages of ampicillin and chloramphenicol in a prospective randomized trial of 36 children and adults with meningitis. The groups were comparable based on clinical, laboratory, and etiological parameters. Ceftriaxone given once daily produced results equivalent to those obtained with ampicillin plus chloramphenicol, as judged by cure rate, case fatality ratio, resolution with sequelae, type and severity of sequelae, time to sterility of cerebrospinal fluid, and potentially drug-related adverse effects. The cerebrospinal fluid bactericidal titers obtained 16 to 24 h after ceftriaxone dosing were usually 1:512 to greater than 1:2,048 even late in the treatment course, compared with values of 1:8 to 1:32 in patients receiving ampicillin plus chloramphenicol. Ceftriaxone clearly deserves further evaluation for the therapy of meningitis; the optimal dose, dosing frequency (every 12 h or every 24 h), and duration of therapy remain to be determined.


Reflex and reflective testing: efficiency and effectiveness of adding on laboratory tests.
Laboratory investigations may be added to existing requests either automatically on the basis of algorithms (reflex testing) or by laboratory professionals (reflective testing). The clinical utility of reflex and reflective testing is not fully established. We studied efficiency (number of tests that needs to be added to make a diagnosis) and effectiveness (number of diagnoses) of reflex and reflective testing in selected biochemical scenarios. Using fixed rules, we prospectively measured efficiency and effectiveness of reflex and reflective testing in the following scenarios (reflex initiators in parentheses): (1) hypovitaminosis D (hypocalcaemia plus elevated alkaline phosphatase activity); (2) hypomagnesaemia (hypokalaemia or hypocalcaemia); (3) hypothyroidism (high thyroid-stimulating hormone [TSH]); (4) hyperthyroidism (low TSH); (5) haemochromatosis (reflex or reflective addition of iron studies, followed by reflective addition of genetic studies). Separately, using a different data-set, we examined the impact of varying TSH thresholds on outcomes in the biochemical diagnosis of hyper- and hypothyroidism. In patients aged over 55 y, 25-hydroxy-vitamin D <50 nmol/L could be predicted with > or =90% certainty when albumin-adjusted calcium was < or =2.1 mmol/L plus alkaline phosphatase >150 U/L. Higher numbers of tests were needed to make a diagnosis in other scenarios. In general, more diagnoses were made by reflex testing. Outside the euthyroid TSH range, efficiency of diagnosis of hyper- and hypothyroidism became asymptotic, while effectiveness declined. Near-maximal efficiency of reflex testing can be achieved, depending on the reflex and diagnostic thresholds applied. Reflective and reflex testing are complementary activities, the clinical utility of which depends on the initiators used.


Sub-ambient pressure IR-MALDI ion mobility spectrometer for the determination of low and high field mobilities.
A new ion mobility (IM) spectrometer, enabling mobility measurements in the pressure range between 5 and 500 mbar and in the reduced field strength range E/N of 5-90 Td, was developed and characterized. Reduced mobility (K0) values were studied under low E/N (constant value) as well as high E/N (deviation from low field K0) for a series of molecular ions in nitrogen. Infrared matrix-assisted laser desorption ionization (IR-MALDI) was used in two configurations: a source working at atmospheric pressure (AP) and, for the first time, an IR-MALDI source working with a liquid (aqueous) matrix at sub-ambient/reduced pressure (RP). The influence of RP on IR-MALDI was examined and new insights into the dispersion process were gained. This enabled the optimization of the IM spectrometer for best analytical performance. While ion desolvation is less efficient at RP, the transport of ions is more efficient, leading to intensity enhancement and an increased number of oligomer ions. When deciding between AP and RP IR-MALDI, a trade-off between intensity and resolving power has to be considered. Here, the low field mobility of peptide ions was first measured and compared with reference values from ESI-IM spectrometry (at AP) as well as collision cross sections obtained from molecular dynamics simulations. The second application was the determination of the reduced mobility of various substituted ammonium ions as a function of E/N in nitrogen. The mobility is constant up to a threshold at high E/N. Beyond this threshold, mobility increases were observed. This behavior can be explained by the loss of hydrated water molecules.


Norepinephrine Inhibits the Proliferation of Human Bone Marrow-Derived Mesenchymal Stem Cells via β2-Adrenoceptor-Mediated ERK1/2 and PKA Phosphorylation.
Bone marrow-derived mesenchymal stem cells (BMSCs) represent an alternative to chondrocytes to support cartilage regeneration in osteoarthritis (OA). The sympathetic neurotransmitter norepinephrine (NE) has been shown to inhibit their chondrogenic potential; however, their proliferation capacity under NE influence has not been studied yet. Therefore, we used BMSCs obtained from trauma and OA donors and compared the expression of adrenergic receptors (AR). Then, BMSCs from both donor groups were treated with NE, as well as with combinations of NE and α1-, α2- or β1/2-AR antagonists (doxazosin, yohimbine or propranolol). Activation of AR-coupled signaling was investigated by analyzing ERK1/2 and protein kinase A (PKA) phosphorylation. A similar but not identical subset of ARs was expressed in trauma (α2B-, α2C- and β2-AR) and OA BMSCs (α2A-, α2B-, and β2-AR). NE in high concentrations inhibited the proliferation of both trauma and OA BMCSs significantly. NE in low concentrations did not influence proliferation. ERK1/2 as well as PKA were activated after NE treatment in both BMSC types. These effects were abolished only by propranolol. Our results demonstrate that NE inhibits the proliferation and accordingly lowers the regenerative capacity of human BMSCs likely via β2-AR-mediated ERK1/2 and PKA phosphorylation. Therefore, targeting β2-AR-signaling might provide novel OA therapeutic options.