Ecotin-like serine peptidase inhibitor ISP1 of Leishmania major plays a role in flagellar pocket dynamics and promastigote differentiation.

Leishmania ISPs are ecotin-like natural peptide inhibitors of trypsin-family serine peptidases, enzymes that are absent from the Leishmania genome. This led to the proposal that ISPs inhibit host serine peptidases and we have recently shown that ISP2 inhibits neutrophil elastase, thereby enhancing parasite survival in murine macrophages. In this study we show that ISP1 has less serine peptidase inhibitory activity than ISP2, and in promastigotes both are generally located in the cytosol and along the flagellum. However, in haptomonad promastigotes there is a prominent accumulation of ISP1 and ISP2 in the hemidesmosome and for ISP2 on the cell surface. An L. major mutant deficient in all three ISP genes (Δisp1/2/3) was generated and compared with Δisp2/3 mutants to elucidate the physiological role of ISP1. In in vitro cultures, the Δisp1/2/3 mutant contained more haptomonad, nectomonad and leptomonad promastigotes with elongated flagella and reduced motility compared with Δisp2/3 populations, moreover it was characterized by very high levels of release of exosome-like vesicles from the flagellar pocket. These data suggest that ISP1 has a primary role in flagellar homeostasis, disruption of which affects differentiation and flagellar pocket dynamics.

Central-line-associated bloodstream infections in a pediatric oncology and hematology hospital at home program.

OBJECTIVE: Central-line-associated bloodstream infections (CLABSIs) are associated with significant morbidity among pediatric oncology-hematology patients, and risk factors remain largely unknown in the setting of hospital at home (HAH). Children in HAH receive intensive treatment (eg, chemotherapy and parenteral nutrition), with frequent central-line handling; thus, they may be at higher risk for CLABSI. METHODS: We conducted a monocentric retrospective study of patients with a central line included in our HAH program from January 1 to December 31, 2016. HAH patient characteristics for children developing CLABSIs were compared to those who did not, based on blood cultures positive for infection and clinical data of all patients included. RESULTS: Overall, 492 HAH stays were analyzed, with 144 patients. The overall CLABSI rate in these patients was 2.6 per 1,000 central-line days. Children who developed CLABSIs were younger (median age, 2.5 vs 8.8 years; P < .001), suffered more from hematological pathologies (malignant or nonmalignant, 75% vs 52%; P = .02), and had more frequently undergone hematopoietic stem-cell transplantation (30.8% vs 6.5%; P = .01). In addition, these patients often had a tunneled externalized catheter as the central line and were more frequently given parenteral nutrition at home (46% vs 8%; P < .001). CONCLUSIONS: CLABSI rates for children in HAH were more similar to those of inpatients than to rates previously reported for ambulatory patients. The factors associated with infection identified herein should be further validated in multicentric studies and considered to improve HAH practices, parallel to prevention measures used in the inpatient setting.

Influence of parasite encoded inhibitors of serine peptidases in early infection of macrophages with Leishmania major.

Ecotin is a potent inhibitor of family S1A serine peptidases, enzymes lacking in the protozoan parasite Leishmania major. Nevertheless, L. major has three ecotin-like genes, termed inhibitor of serine peptidase (ISP). ISP1 is expressed in vector-borne procyclic and metacyclic promastigotes, whereas ISP2 is also expressed in the mammalian amastigote stage. Recombinant ISP2 inhibited neutrophil elastase, trypsin and chymotrypsin with K(i)s between 7.7 and 83 nM. L. major ISP2-ISP3 double null mutants (Deltaisp2/3) were created. These grew normally as promastigotes, but were internalized by macrophages more efficiently than wild-type parasites due to the upregulation of phagocytosis by a mechanism dependent on serine peptidase activity. Deltaisp2/3 promastigotes transformed to amastigotes, but failed to divide for 48 h. Intracellular multiplication of Deltaisp2/3 was similar to wild-type parasites when serine peptidase inhibitors were present, suggesting that defective intracellular growth results from the lack of serine peptidase inhibition during promastigote uptake. Deltaisp2/3 mutants were more infective than wild-type parasites to BALB/c mice at the early stages of infection, but became equivalent as the infection progressed. These data support the hypothesis that ISPs of L. major target host serine peptidases and influence the early stages of infection of the mammalian host.

Intra peritoneal abdominal fat area measured from computed tomography is an independent factor of severe acute pancreatitis.

PURPOSE: The purpose of this study was to search for a possible relationship between acute pancreatitis (AP) severity and visceral fat (VF) surface on contrast-enhanced computed tomography (CECT). MATERIAL AND METHOD: A total of 112 patients with AP who underwent CECT within 2 to 3 days after the beginning of AP were included. There were 68 mean and 44 women, with a mean age of 56.3±21.6 (SD) years (range: 19-98 years). AP was regarded as mild for patients with an hospital stay up to 5 days and severe for those with an hospital stay greater than 5 days. VF surface was measured on CECT at the level of L4-L5 and of the umbilicus. Association between AP severity and VF surface, computed tomography severity index (CTSI), modified CTSI (mCTSI) and other variables were searched for using uni- and multivariate analysis. RESULTS: At univariate analysis, the VF surface at the level of L4 was greater in patients with severe AP (129.3±68.6 [SD] cm2; range: 21.8-355.8 cm2) than in patients with mild AP (100.1±68.4 [SD] cm2; range:13.2-333 cm2) (P=0.006). Similarly, the VF surface at the umbilicus was greater in patients with severe AP (161.1±76.1 [SD] cm2; range: 31.3-376.7cm2) than in those with mild AP (128.4±74.3cm2; range: 12.8-323.1cm2) (P=0.024). CTSI and mCTSI were also associated to AP severity. At multivariate analysis, only VF surface either measured at the umbilical or at the L4-L5 level was associated with AP severity (P=0.017 and 0.006, respectively). CONCLUSION: VF surface at the level of L4-L5 on CECT is an independent factor of AP severity. VF surface at the level of L4-L5 on CECT is an independent factor of AP severity. These results are in line with recent data on the role of abdominal fat in the genesis of inflammatory response, which is associated with severe forms of AP.

Scanographic features of gastrointestinal stromal tumors.

PURPOSE: To present the scanographic features of gastrointestinal stromal tumor (GIST) and to discuss their differential diagnosis. PATIENTS AND METHODS: A retrospective study of 45 patients who underwent surgery for GIST between January 1990 and March 2006 was performed. RESULTS: Patient age was 64 years on average. The most common symptoms were abdominal pain and gastrointestinal bleeding. Tumors were located in the stomach in 28 patients (body: 19, antrum: 5, fundus: 4), the small intestine in 13 (jejunum: 6, duodenum: 4, ileum: 3), the rectum in two and the small bowel mesentery in two. Computed tomography showed a large (average size: 9.2 cm, range 3.3-30 cm) exophytic extragastric lobulated mass with an associated wall thickening in 35 cases (78%). The pattern was an endoluminal polyp (average size: 3.2 cm, range 2.2-5.5 cm) in eight cases (18%). The two mesenteric stromal tumors (4%) were seen as well-delimited lobulated large masses (3 and 12 cm). The enhancement was peripheral with central hemorrhagic, necrotic and cystic areas in 37 cases (82%). Mucosal ulceration was seen in 18 cases (40%) and ascites in five (11%). Peritoneal spread and liver metastasis were demonstrated in three patients (7%). Calcification, metastatic lymphadenopthy, venous thrombosis or vascular invasion were not seen. CONCLUSION: Scanographic features of GIST can suggest the diagnosis of GIST before surgery.

PFPI-like genes are expressed in Leishmania major but are pseudogenes in other Leishmania species.

Pyrococcus furiosus protease I (PFPI) is a multimeric cysteine peptidase from P. furiosus. Genome analyses indicate that orthologues are present in rather few other organisms, including Dictyostelium discoideum and several bacteria, Archaea and plants. An open reading frame (ORF) coding for a PFPI-like protein (PFP1) was identified in Leishmania major and Leishmania mexicana and full-length spliced and polyadenylated PFP1 mRNA detected for both species. Vestiges of a PFPI-like gene could also be identified in Leishmania braziliensis and Leishmania infantum, but no ORF remains owing to the presence of frame-shifts and stop codons. No evidence for a PFPI-like gene could be found in the syntenic region of Trypanosoma brucei or Trypanosoma cruzi, raising the possibility that the PFPI-like genes were acquired by a lateral gene transfer event after the divergence of trypanosomes and Leishmania. The gene may have subsequently degenerated into a pseudogene in some Leishmania species, owing to the loss of relevant biological function. However, antibodies raised against L. mexicana recombinant protein detected PFP1 in promastigote extracts of L. major, but not in L. mexicana promastigote or amastigote extracts. The expression of PFP1 in L. major suggests that PFP1 might contribute to the disease tropism that distinguishes this Leishmania species from others.

Equal variations of the Fermi level and work function in graphene at the nanoscale.

If surface effects are neglected, any change of the Fermi level in a semiconductor is expected to result in an equal and opposite change of the work function. However, this is in general not observed in three-dimensional semiconductors, because of Fermi level pinning at the surface. By combining Kelvin probe force microscopy and scanning tunneling spectroscopy on single layer graphene, we measure both the local work function and the charge carrier density. The one-to-one equivalence of changes in the Fermi level and the work function is demonstrated to accurately hold in single layer graphene down to the nanometer scale.

An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum.

An NAD(+)-dependent glutamate dehydrogenase (GDH; EC 1.4.1.24) was cloned from the ruminal ciliate protozoan, Entodinium caudatum. The gene had high sequence similarity to GDH genes from the Bacteroides (class)--a class of bacteria which is highly represented in the rumen. When expressed in Escherichia coli the enzyme had a high affinity for ammonia and alpha-ketoglutarate (apparent K(m) of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent K(m) of 98 mM). GDH activity and GDH mRNA concentration were increased by incubating washed E. caudatum cells with ammonia and antibiotics. These results suggest that the GDH is an anabolic enzyme catalysing the assimilation of ammonia by E. caudatum in the rumen and that the gene was probably acquired by lateral gene transfer from a ruminal bacterium.

Cognitive and behavioral treatment of pathological gambling: a controlled study.

This study evaluated the efficacy of a cognitive-behavioral treatment package for pathological gambling. Twenty-nine men who met criteria for pathological gambling in accordance with the Diagnostic and Statistical Manual of Mental Disorders (3rd ed., revised; DSM-III-R; American Psychiatric Association, 1987) were randomly assigned to treatment or wait-list control. The treatment included 4 components: (a) cognitive correction of erroneous perceptions about gambling, (b) problem-solving training, (c) social skills training, and (d) relapse prevention. The dependent variables were the South Oaks Gambling Screen, perception of control, frequency of gambling, perceived self-efficacy, desire to gamble, and number of DSM-III-R criteria met by participants. Posttest results indicated highly significant changes in the treatment group on all outcome measures, and analysis of data from 6- and 12-month follow-ups revealed maintenance of therapeutic gains. Recommendations for clinical interventions are discussed and focus on the cognitive correction of erroneous perceptions of gambling.

Cognitive treatment of pathological gamblers.

This study evaluates the efficacy of a cognitive treatment for pathological gambling. Five pathological gamblers were treated in a multiple baseline across subjects design. Cognitive correction targeted the erroneous perceptions towards the notion of randomness. Four subjects reported a clinically significant decrease in the urge to gamble, an increase in their perception of control, and no longer met the DSM-IV criteria for pathological gambling. Therapeutic gains were maintained at the 6 month follow-up. Results suggest that cognitive therapy targeting the misconception of the notion of randomness is a promising treatment for pathological gambling, a refractory disorder to most therapeutic interventions.

Group therapy for pathological gamblers: a cognitive approach.

This study evaluated the efficacy of a group cognitive treatment for pathological gambling. Gamblers, meeting DSM-IV criteria for pathological gambling, were randomly assigned to treatment (N=34) or wait-list control (N=24) conditions. Cognitive correction techniques were used first to target gamblers' erroneous perceptions about randomness, and then to address issues of relapse prevention. The dependent measures used were the DSM-IV criteria for pathological gambling, perceived self-efficacy, gamblers' perception of control, desire to gamble, and frequency of gambling. Post-treatment results indicated that 88% of the treated gamblers no longer met the DSM-IV criteria for pathological gambling compared to only 20% in the control group. Similar changes were observed on all outcome measures. Analysis of data from 6-, 12- and 24-month follow-ups revealed maintenance of therapeutic gains. Recommendations for group interventions are discussed, focusing on the cognitive correction of erroneous perceptions toward the notion of randomness.

Treatment of pathological gamblers: an experimental study.

The efficacy of a behavioral treatment for pathological gamblers, including cognitive interventions, problem solving and relapse prevention is examined. Three pathological gamblers meeting DSM-III-R criteria were studied in an experimental multiple baseline design. By the end of treatment the subjects ceased all gambling behaviors, increased their perception of self-control of gambling, and evaluated their problems as less severe. Therapeutic gains were maintained at 6- and 9-month follow-ups. The importance of problem solving and the modification of erroneous perceptions in treatment programs for pathological gambling is discussed.

[Fatal fulminant hepatitis in 2 women taking toloxatone (Humoryl)]. / Hépatites fulminantes mortelles chez deux femmes prenant de la toloxatone (Humoryl).

We report two cases of fulminant hepatitis which might be due to toloxatone, a new type-A monoamine oxidase inhibitor. Hepatitis occurred 20 days after the beginning of toloxatone administration in the first case and 138 days after the reintroduction of treatment in the second case. Clinical features included vomiting and jaundice, followed by asterixis and coma. Histologically, hepatic cell necrosis was predominant in the centrilobular area in the first case, and affected the entire lobule in the second case. Both patients died despite emergency liver transplantation.

Variations in internet-related problems and psychosocial functioning in online sexual activities: implications for social and sexual development of young adults.

This survey of 760 university students examined differences in Internet-related problems and psychosocial functioning between four patterns of participation in online sexual information and entertainment activities. Students who did not participate in either online sexual activity were more satisfied with their offline life and more connected to friends and family. Those who engaged in both online sexual activities were more dependent on the Internet and reported lower offline functioning. Students who only sought sexual information maintained strong offline affiliations. Those who only sought entertainment did not report lower offline functioning. Respondents most deficient in offline social support did not report compensatory online support. Despite students' common participation in online sexual activities (OSA) as a venue for social and sexual development, those relying on the Internet and the affiliations it provides appear at risk of decreased social integration. The authors discussed the finding's implications for social and sexual development.

[Schizophrenia, special approach and continuity of care : The first episode program of Hôtel-Dieu Hospital in Lévis, Québec.]. / Schizophrénie, approche spécialisée et continuité de soins. Le programme spécifique d'intervention Premier-Episode de l'Hôtel-Dieu de Lévis.

Schizophrenia is a complex illness with an evolutive character. Based on a conceptual framework of cognitive orientation, the specific intervention program First Episode of Hôtel-Dieu in Lévis includes a complete and standardized assessment to an individualized and family plan. Different methods of treatment acording to an individualized approach (psyhco-education, psychotherapy) as well as group therapy (psychological intervention at the cognitive level or Brenner's Integrated psychological therapy) are then proposed. Psycho-education intervention for families is also offered. Structures and different steps described here, harmonize with those already in place before the program's creation thus offering a continuity in care. The underlying conceptual framework and the different methods of functioning of the program are also presented.

Loss of secretory pathway FK506-binding proteins results in cold-sensitive lethality and associate extracellular matrix defects in the nematode Caenorhabditis elegans.

The FK506-binding proteins (FKBs) represent ubiquitous enzymes that catalyze the rate-limiting peptidyl prolyl cis-trans isomerization step in protein folding. The nematode Caenorhabditis elegans has eight FKBs, three of which (FKB-3, -4, and -5) have dual peptidyl prolyl cis-trans isomerase (PPIase) domains, signal peptides and ER retention signals. PPIase activity has been detected for recombinant FKB-3. Both FKB-3 and -5 are expressed in the exoskeleton-synthesizing hypodermis with transcript peaks that correspond to the molting and collagen synthesis cycles. FKB-4 is expressed at a low level throughout development. No phenotypes were observed in deletion mutants in each of the secretory pathway FKBs. Combined triple and fkb-4, -5 double deletion mutants were however found to arrest at 12 degrees C, but developed normally at 15-25 degrees C. This cold-sensitive larval lethal effect was not maternally derived, occurred during embryogenesis, and could be rescued following the transgenic introduction of a wild type copy of either fkb-4 or fkb-5. The temperature-sensitive defects also affected molting, cuticle collagen expression, hypodermal seam cell morphology, and the structural integrity of the cuticular extracellular matrix. This study establishes that the secretory pathway FK506-binding PPIase enzymes are essential for normal nematode development, collagen biogenesis, and the formation of an intact exoskeleton under adverse physiological conditions.

Social cost of pathological gambling.

Pathological gambling creates enormous problems for the afflicted individuals, their families, employers, and society, and has numerous disastrous financial consequences. The present study evaluates the financial burdens of pathological gambling by questioning pathological gamblers in treatment in Gamblers Anonymous (n=60; 56 males, 4 females; mean age = 40 years old) about personal debts, loss of productivity at work, illegal activities, medical costs and the presence of other dependencies. Results show that important debts, loss of productivity at work and legal problems are associated with pathological gambling. Discussion is formulated in terms of the social cost of adopting a liberal attitude toward the legalization of various gambling activities.

Cognitive treatment of pathological gambling.

This study evaluated the efficacy of a cognitive treatment package for pathological gambling. Sixty-six gamblers, meeting DSM-IV criteria for pathological gambling, were randomly assigned to treatment or wait-list control conditions. Cognitive correction techniques were used first to target gamblers' erroneous perceptions about randomness and then to address issues of relapse prevention. The dependent measures used were the South Oaks Gambling Screen, the number of DSM-IV criteria for pathological gambling met by participants, as well as gamblers' perception of control, frequency of gambling, perceived self-efficacy, and desire to gamble. Posttest results indicated highly significant changes in the treatment group on all outcome measures, and analysis of data from 6- and 12-month follow-ups revealed maintenance of therapeutic gains. Recommendations for clinical interventions are discussed, focusing on the cognitive correction of erroneous perceptions toward the notion of randomness.

The Caenorhabditis elegans ERp60 homolog protein disulfide isomerase-3 has disulfide isomerase and transglutaminase-like cross-linking activity and is involved in the maintenance of body morphology.

A novel protein disulfide isomerase gene, pdi-3, was isolated from the nematode Caenorhabditis elegans. This gene encodes an enzyme related to the ERp60 class of thioredoxin proteins and was found to exhibit unusual enzymatic properties. Recombinant protein displayed both disulfide bond isomerase activity and calcium-dependent transglutaminase-like cross-linking activity. The pdi-3 transcript was developmentally constitutively expressed, and the encoded protein is present in many tissues including the gut and the hypodermis. The nematode hypodermis synthesizes the essential collagenous extracellular matrix (ECM) called the cuticle. Transcript disruption via double-stranded RNA interference resulted in dramatic and specific synthetic phenotypes in several C. elegans mutant alleles with weakened cuticles: sqt-3(e2117), dpy-18(e364, ok162, and bx26). These nematodes displayed severe dumpy phenotypes and disrupted lateral alae, a destabilized cuticle and abnormal male and hermaphrodite tail morphologies. These defects were confirmed to be consistent with hypodermal seam cell abnormalities and corresponded with the severe disruption of a cuticle collagen. Wild type nematodes did not exhibit observable morphological defects; however, cuticle collagen localization was mildly disrupted following pdi-3 RNA interference. The unusual thioredoxin enzyme, protein disulfide isomerase-3, may therefore play a role in ECM assembly. This enzyme is required for the proper maintenance of post-embryonic body shape in strains with a weakened cuticle, perhaps through ECM stabilization via cross-linking activity, disulfide isomerase protein folding activity, protein disulfide isomerase chaperone activity, or via multifunctional events.