The relationship between non-purging compensatory behaviors, clinical severity, and treatment outcomes in adults with binge-spectrum eating disorders.

Non-purging compensatory behaviors (NPCB; e.g. driven exercise, fasting, other extreme behaviors) are a subcategory of compensatory behaviors typically characterized as infrequent and less severe. Limited prior research has studied NPCB despite their increasing prevalence among adults with binge-spectrum eating disorders (B-ED). More research is needed to understand the types of NPCB present among B-ED and the association between NPCB, clinical severity, and treatment outcomes. Secondary analyses were conducted among 155 adults with B-ED in cognitive-behavioral (CBT)-based clinical trials. At baseline and post-treatment, clinical interviews of eating pathology assessed binge eating frequency, purging compensatory behavior frequency, and global eating pathology. The following NPCB were also assessed: driven exercise, 24-h fasting, 8+ waking hours of compensatory fasting, chewing and spitting, and other extreme weight control behaviors. Participants engaging in NPCB reported higher global eating pathology than those not engaging in NPCB. Frequency of chewing and spitting and 24-h fasting significantly decreased over treatment. Engagement in NPCB at baseline did not predict CBT outcomes. The current study highlights the prevalence and clinical severity of NPCB in B-ED but offers promising results regarding the potential for CBT to improve these behaviors. More research is needed on other extreme weight control behaviors reported qualitatively in our sample and on the maintenance of improvements in non-purging behaviors after CBT.

Comparison of oral fluid collection methods for the molecular detection of hepatitis B virus.

BACKGROUND: This study aims to compare the efficiency of four oral fluid collection methods (Salivette, FTA Card, spitting and DNA-Sal) to detect HBV DNA by qualitative PCR. MATERIALS AND METHODS: Seventy-four individuals (32 HBV reactive and 42 with no HBV markers) donated serum and oral fluid. In-house qualitative PCR to detect HBV was used for both samples and commercial quantitative PCR for serum. RESULTS: HBV DNA was detected in all serum samples from HBV-infected individuals, and it was not detected in control group. HBV DNA from HBV group was detected in 17 samples collected with Salivette device, 16 samples collected by FTA Card device, 16 samples collected from spitting and 13 samples collected by DNA-Sal device. Samples that corresponded to a higher viral load in their paired serum sample could be detected using all oral fluid collection methods, but Salivette collection device yielded the largest numbers of positive samples and had a wide range of viral load that was detected. CONCLUSION: It was possible to detect HBV DNA using all devices tested, but higher number of positive samples was observed when samples were collected using Salivette device, which shows high concordance to viral load observed in the paired serum samples.

Dried blood spot samples: optimization of commercial EIAs for hepatitis C antibody detection and stability under different storage conditions.

This study was undertaken to optimize and compare the efficiency of two commercial EIAs for anti-HCV detection (HCV Ab Radim, Pomezzia, Italy and ETI-AB-HCVK-4 DiaSorin, Vercelli, Italy), in dried blood spot (DBS) samples. The long-term stability of anti-HCV on DBS samples stored at three environmental conditions was also evaluated at: 2-8 °C, 20-25 °C, and -20 °C. Paired DBS and serum samples were obtained from individuals with or without anti-HCV. The type of elution buffer, sample and conjugate volume, sample incubation time and cut-off values were evaluated. For both EIAs, a larger sample volume was used, and the cut-off value determined by the manufacturer was employed for Radim EIA; however, ROC curve analysis was used for the DiaSorin EIA. The sensitivity and specificity of Radim EIA on DBS were 97.5% and 99.5%, respectively, and of DiaSorin EIA were 88.9% and 98.9%, respectively. Accurate results were obtained for a period of 117 days using DBS samples stored at all storage conditions, but storage at -20 °C resulted in the lowest variation among the absorbance values. Both EIAs demonstrated the same limit of detection (until dilution of 1:10(4) with estimated viral load of 3.1 × 10(-1) UI/ml), but the Radim EIA was associated with the best performance because a low coefficient of variation was observed in the repetition and reproducibility studies. In conclusion, commercial EIAs can be optimized for anti-HCV detection in DBS samples that are extremely stable at different conditions for more than 100 days.

An evaluation of different saliva collection methods for detection of antibodies against hepatitis C virus (anti-HCV).

BACKGROUND: Saliva samples can be used as an alternative fluid for against hepatitis C virus (anti-HCV) detection owing to the ease of collection and excellent acceptability. This study was conducted to optimize a commercial enzyme immunoassay (EIA) to detect anti-HCV in saliva samples. METHODS: Ninety-six individuals donated paired serum and saliva samples that were obtained, using a commercial device (Salivette) and spitting into a sterile container. Initially, elution buffer for the Salivette samples, sample volume, incubation time and temperature, and two different anti-HCV EIAs were evaluated. Using the optimized assay, three methods for cut-off calculation were also evaluated. RESULTS: A 20-fold increase in the sample volume for both collection methods was needed. Moreover, the Radim assay was the most appropriate assay for anti-HCV detection in saliva samples, and the quality parameters were increased when a ROC curve was used to determine the cut-off value. Using this optimized assay, the sensitivities, specificities, accuracies, positive and negative predictive values were above 90% for saliva obtained using both the Salivette and spitting methods. Using this assay, discordant false-negative results were obtained for only two Salivette samples and five spitting samples. The concordance kappa was 93% for the Salivette method and 86.1% for the spitting method, demonstrating excellent performance. CONCLUSIONS: Saliva samples obtained for both methods can be employed for anti-HCV detection among HCV-infected or HCV-suspected cases, but several modifications must be performed on commercial EIAs to obtain good results. Moreover, samples obtained with commercial devices are more appropriate for anti-HCV detection in saliva samples.

Genotyping hepatitis C virus from hemodialysis patients in Central Brazil by line probe assay and sequence analysis.

The present study examined the distribution of hepatitis C virus (HCV) genotypes and subtypes in a hemodialysis population in Goiás State, Central Brazil, and evaluated the efficiency of two genotyping methods: line probe assay (LiPA) based on the 5' noncoding region and nucleotide sequencing of the nonstructural 5B (NS5B) region of the genome. A total of 1095 sera were tested for HCV RNA by RT-nested PCR of the 5' noncoding region. The LiPA assay was able to genotype all 131 HCV RNA-positive samples. Genotypes 1 (92.4%) and 3 (7.6%) were found. Subtype 1a (65.7%) was the most prevalent, followed by subtypes 1b (26.7%) and 3a (7.6%). Direct nucleotide sequencing of 340 bp from the NS5B region was performed in 106 samples. The phylogenetic tree showed that 98 sequences (92.4%) were classified as genotype 1, subtypes 1a (72.6%) and 1b (19.8%), and 8 sequences (7.6%) as subtype 3a. The two genotyping methods gave concordant results within HCV genotypes and subtypes in 100 and 96.2% of cases, respectively. Only four samples presented discrepant results, with LiPA not distinguishing subtypes 1a and 1b. Therefore, HCV genotype 1 (subtype 1a) is predominant in hemodialysis patients in Central Brazil. By using sequence analysis of the NS5B region as a reference standard method for HCV genotyping, we found that LiPA was efficient at the genotype level, although some discrepant results were observed at the subtype level (sensitivity of 96.1% for subtype 1a and 95.2% for subtype 1b). Thus, analysis of the NS5B region permitted better discrimination between HCV subtypes, as required in epidemiological investigations.

Co-circulation of genotypes IA and IB of hepatitis A virus in Northeast Brazil.

The Northeast region is the location of most cases of acute hepatitis A virus (HAV) in Brazil. In the present study, the genotypes of HAV strains from Pernambuco State, one of most populous states in the Northeast region, were characterized. Blood samples positive for anti-HAV IgM from 145 individuals (mean age = 29.1 years), collected during 2002 and 2003, were submitted to nested RT-PCR for amplification of the 5'non-translated region (5'NTR) and VP1/2A regions of the HAV genome. The VP1/2A and 5'NTR regions were amplified in 39 and 21% of the samples, respectively. Nucleotide sequencing was carried out in 46% of VP1/2A and in 53% of 5'NTR isolates. The identity in nucleotide sequence of the VP1/2A region ranged from 93.6 to 100.0%. Phylogenetic analysis of the VP1/2A sequences showed that 65% belong to sub-genotype IA and 35% to sub-genotype IB. Co-circulation of both sub-genotypes was observed in the two years studied. Distinct clusters of highly related sequences were observed in both sub-genotypes, suggesting endemic circulation of HAV strains in this area. In the 5'NTR isolates, 92.7-99.2% identity was observed and two isolates presented one deletion at position 413. Phylogenetic analysis showed that genotype IA strains cluster in the tree in the same way as genotype IB strains, but one IIIA isolate from Spain clusters with genotype IB strains. These results do not allow us to state that 5'NTR could be used to genotype HAV sequences. This is the first report of co-circulation of sub-genotypes IA and IB in this region, providing additional information about the molecular epidemiology of HAV strains in Brazil.

CD95 and TRAIL receptor-mediated activation of protein kinase C and NF-kappaB contributes to apoptosis resistance in ductal pancreatic adenocarcinoma cells.

The molecular alterations in tumour cells leading to resistance towards apoptosis induced by CD95 and TRAIL-receptors are not fully understood. We report here that the stimulation of the CD95- and TRAIL-resistant human pancreatic adenocarcinoma cell line PancTuI with an agonistic anti-CD95 antibody or TRAIL resulted in activation of protein kinase C and NF-kappaB. Inhibition of protein kinase C by Gö6983 sensitized these cells to apoptotic challenges and strongly diminished activation of NF-kappaB by anti-CD95 and TRAIL. Similarly, inhibition of NF-kappaB by MG132 or by transient transfection with a dominant negative mutant of IkappaBalpha restored the responsiveness of PancTuI cells to both death ligands. In the CD95 and TRAIL-sensitive cell line Colo357 the induction of protein kinase C and NF-kappaB following activation of CD95 and TRAIL-R was very moderate compared with PancTuI cells. However, pre-incubation of these cells with PMA strongly reduced their apoptotic response to anti-CD95 and TRAIL. Taken together, we show that activation of protein kinase C operates directly in a death receptor-dependent manner in PancTuI cells and protect pancreatic tumour cells from anti-CD95 and TRAIL-mediated apoptosis by preventing the loss DeltaPsim and Cytochrome c release as well as by induction of NF-kappaB.

Hepatitis B virus DNA stability in plasma samples under short-term storage at 42°C.

We evaluated the stability of hepatitis B virus (HBV) DNA in plasma samples stored at 42°C for external quality assessment (EQA) panels of viral load. To assess the stability of plasma samples containing different concentrations of HBV DNA, serial dilutions of HBV-infected samples with a viral load of 6.40 log(10) IU/mL were made to yield viral loads of 5, 4, and 3 log(10) IU/mL. These were incubated at 42°C for up to 7 days and then frozen at -70°C. Viral load testing for HBV DNA was performed for all samples using COBAS® AmpliPrep/COBAS® TaqMan® HBV Test (v.2.0, Roche, Switzerland). Results were compared with fresh frozen plasma samples as a benchmark to establish acceptable measurements on the days following sample collection. Although the results of this study demonstrated a decrease in HBV DNA viral load ranging from 0.005 to 0.30 log(10) IU/mL after storage at 42°C for up to 7 days, these values did not exceed 0.5 log(10), which is the estimated intra-assay variation for molecular tests. Thus, the insignificant decrease in viral load suggests that shipment of HBV in plasma samples at temperatures of up to 42°C is permissible if they are frozen within 7 days.

Management of arterial puncture site after catheterization procedures: evaluating a suture-mediated closure device.

To overcome the challenge associated with achievement in hemostasis after a catheterization procedure, a suture-based closure device was compared with manual compression in a 600-patient randomized trial. The major study end points included the incidence of vascular complications and the time to ambulation after the procedure. The study included diagnostic or interventional procedures. The suture-mediated closure was performed immediately after the procedure independent of the anticoagulation level, whereas manual compression was performed per hospital protocol with sheath removal relying on normalization of patient's anticoagulation status. A significant reduction in time to achieve hemostasis (7.8 +/- 4.8 vs 19.6 +/- 13.2 minutes, p <0001) and time to ambulation (4.5 +/- 6.5 vs 17.8 +/- 5 hours, p <0001) was associated with use of the suture-mediated closure device. The incidence of vascular complications was similar in the overall population (5.7% for suturing device vs 11.3% for compression) or in the interventional patient subset (8.4% for suturing device vs 9.6% for compression). There was a significant reduction in the incidence of vascular complications in the diagnostic procedure subset (4.4% for suturing device vs 12.1% for compression, p <0.05). Thus, the use of a suture-mediated closure device represents a safe alternative to manual compression. Hemostasis and ambulation can be achieved faster with the suturing device than with manual compression, with a potential reduction in access site complications.

Autotransplantation of porcine islets of Langerhans.

Autotransplantation of the islets of Langerhans in pancreatic tissue fragments, prepared with no attempt to separate exocrine tissue, produced viable, well vascularized implants in the peritoneal cavity of pigs. We autografted 31 pancreatectomized pigs with pancreatic preparations in which exocrine tissue either was separated completely, partially, or not at all. Nine pancreatectomixed pigs were used as control animals. On the basis of the results of the standard serum glucose and insulin level tests and the mean survival results, the various methods of tissue preparation had no discernible effect. One month after grafting, the implants from the unseparated pancreatic preparations contained significant amounts of insulin and cells identical to normal beta cells. One pig with a free-fragment autograft is alive and well, with an increase in the size of its implants, 6 months later. Separation of pancreatic endocrine and exocrine tissue is not essential for autografting the islets of Langerhans in the pig. The exocrine tissue apparently can be removed by in vivo mechanisms, and the endocrine tissue is preserved in free-fragment pancreatic grafts.

Dengue haemorrhagic fever in the state of Rio de Janeiro, Brazil: a study of 56 confirmed cases.

We studied 56 cases of serologically confirmed dengue haemorrhagic fever living in the metropolitan area of Niterói and surrounding cities in the state of Rio de Janeiro, Brazil. The most frequent findings were fever and myalgia. Spontaneous haemorrhagic manifestations occurred in 46 patients, and 23 of these had more than one kind of bleeding; petechiae and bleeding gums were the most frequent association. The distribution according to the World Health Organization's criteria of severity was 6 in grade I, 23 in grade II, 24 in grade III and 3 in grade IV.

Genetic variability of hepatitis A virus isolates in Rio de Janeiro: implications for the vaccination of school children.

The epidemiology of hepatitis A virus (HAV) infection is shifting from high to intermediate endemicity in Brazil, resulting in increased numbers of susceptible individuals and a greater potential for the emergence of outbreaks. Universal vaccination against HAV has been recommended for children, but updated sero-epidemiological data are necessary to analyze the level of natural immunity and to identify candidates for preventive measures. In addition, more molecular studies are necessary to characterize the genotypes involved in HAV infections and outbreaks. Sera from 299 school children (5-15 years old) and 25 school staff members, collected during an outbreak of HAV at a rural public school in June 2000, were tested for IgM and total anti-HAV antibodies (ELISA). Viral RNA was amplified by RT-PCR from anti-HAV IgM-positive sera and from 19 fecal samples. Direct nucleotide sequencing of the VP1/2A region was carried out on 18 PCR-positive samples. Acute HAV infection was detected by anti-HAV IgM in 93/299 children and in 3/25 adult staff members. The prevalence of total anti-HAV antibodies in IgM-negative children under 5 years of age was only 10.5%. HAV-RNA was detected in 46% IgM-positive serum samples and in 16% stool samples. Sequence analysis showed that half the isolates belonged to subgenotype IA and the other half to IB. On the basis of these data, mass vaccination against HAV is recommended without prevaccination screening, especially for children before they enter school, since nearly 90% of the children under 5 years were susceptible. Molecular characterization indicated the endemic circulation of specific HAV strains belonging to subgenotypes IA and IB.

Human antibodies to dengue and yellow fever do not react in diagnostic assays for hepatitis C virus.

Hepatitis C virus (HCV) is a recently described causative agent of the great majority of post-transfusion non A-non B hepatitis and is classified within the Flaviviridae family. Due to a high prevalence of anti-HCV and other flaviviruses circulating in Brazil, such as dengue and yellow fever, we investigated the possibility of serological cross-reactivity between these viruses. Different panels of human sera positive for dengue type 1 (9 cases) and type 2 (7 cases) from 6 patients naturally infected with yellow fever and from 94 adults vaccinated against the 17D strain of yellow fever were tested against HCV antigens used in diagnostic assays. Two enzyme immunoassay systems were tested: one, an in-house test using recombinant antigens from core, NS3 and NS5 regions of the HCV genome (Research Foundation for Microbial Disease of Osaka University, Japan); and another, using synthetic peptides representing immunodominant epitopes of structural core and non-structural NS4 and NS5 HCV regions (INNOTEST HCV Ab, Innogenetics, Belgium). A line immunoassay (INNO-LIA HCV Ab, Innogenetics, Belgium) was used as a confirmatory test. In this, HCV antigens are coated as discrete lines on a nylon strip with plastic backing. Besides 4 control lines on each strip, a total of 6 HCV lines are present: line A consists of several NS4 epitopes, line B consists of several NS5 epitopes and lines C-F contain several core epitopes. This test not only confirms but differentiates antibodies to hepatitis C virus. No positive results were detected with these tests, indicating that hepatitis C infection can be evaluated by current assays in regions where flaviviruses are endemic.

Oligosaccharides and derivatives--integrating biocatalyst selectivity and chemical diversity.

Several biocatalytic pathways are available to synthesize rare sugars, oligosaccharides and derivatives. As examples selective hydrolysis, transglycosylation and regioselective oxidation are presented illustrating their potential. From inulin, difructose anhydrides and inulobiose are accessible via specific hydrolases with transglycosylation activity. Glycosyltransferases of the sucrase type utilise sucrose as a versatile substrate for the selective transfer of either glucosyl- or fructosyl units to a broad range of acceptors, sugars as well as derivatives, to yield oligosaccharides of different type. Kinetic analysis and reaction engineering can provide high product yields and concentration, as will be shown for glucose, yielding isomaltooligosaccharides as an example. Several new acceptor saccharides, including sugar alcohols and acids, have been shown to give new oligosaccharides. Regioselective oxidation of disaccharides like sucrose, lactose and others with resting cells of Agrobacterium tumefaciens yield 3-keto-disaccharides. These can further be functionalized in aquous systems without protecting groups via established chemical routes, such as catalytic hydration. As an example allosucrose is thus obtained from 3-keto-sucrose with high yield and stereochemical selectivity, which in turn can easily be hydrolysed and separated from fructose to give allose. Amino acid and peptide conjugates are accessible via reductive amination and acylation, as are building blocks for polymerisation.

Simultaneous detection of hepatitis C virus antigen and antibodies in dried blood spots.

BACKGROUND: Enzyme immunoassays (EIA) designed to detect hepatitis C virus (HCV) core antigen and anti-HCV antibodies (HCV AgAb) simultaneously can improve the early detection of HCV infection when molecular diagnostic methods are not widely available. OBJECTIVES: To evaluate the suitability of dried blood spot (DBS) samples for detecting HCV AgAb using commercial EIAs. STUDY DESIGN: Paired serum and DBS samples were assayed using two commercial EIAs for HCV AgAb (Monolisa™ HCV AgAb ULTRA and Murex HCV AgAb). Manufacturer's recommendations were followed for sera while sample volume, incubation time and cut-off (CO) determination were evaluated for the DBS samples. The values of sensitivity, specificity, inter-rater agreement, detection limit, assay precision and stability of DBS samples at different conditions (22-26°C, 2-8°C and -20°C) were determined. RESULTS: It was necessary to increase the DBS sample volume fourfold compared to the sera samples to approximate the DBS Optical Density (OD) values to the sera OD values. Using ROC curve to recalculate CO values for the DBS samples, sensitivity was 97.5% for both EIAs, while the specificity was 99.71% for Monolisa™ HCV AgAb ULTRA and 95.95% for Murex HCV AgAb. Accurate testing results were obtained with DBS samples for 60 days at all conditions evaluated; storage at -20°C resulted in low OD variation. Both EIAs demonstrated the same limit of detection among DBS samples [estimated viral load of 3.1 International Units per millilitre (IU/mL)] and low OD value variability in repetitivity and reproducibility studies. CONCLUSION: DBS samples can be used for the detection of HCV AgAb by EIA as they present comparable performance characteristics and excellent stability among various storage conditions.

Hepatitis C virus infection in patients with tuberculosis in Central Brazil.

SETTING: Goiânia City, Goiás State, Brazil. OBJECTIVES: To determine the prevalence of hepatitis C virus (HCV) infection, risk factors, HCV genotype/ subtype, HCV viral load and human immunodeficiency virus (HIV) status in patients with tuberculosis (TB) in Central Brazil. DESIGN: A cross-sectional study was carried out with 402 patients who were under tuberculosis (TB) treatment in the reference hospital for infectious diseases in Goiânia, Goiás, Central Brazil. RESULTS: The prevalence rates of HCV and HIV were respectively 7.5% and 27.6%. Two thirds of the HCV-infected patients (20/30) were HIV-positive. Age, injecting drug use (IDU) and HIV status were factors independently associated with HCV infection. HCV RNA was detected in 23 serum samples; HCV RNA levels were measured in 22/23 samples. HCV RNA level was slightly higher in HCV-HIV co-infected patients than in HCV monoinfected patients. Genotypes 1 (n = 17) and 3 (n = 6) were determined by LiPA. Using phylogenetic tree analysis of the NS5B region, subtypes 1a (n = 12), 1b (n = 2) and 3a (n = 6) were identified. CONCLUSION: These data indicate that patients with TB may benefit from integrated HIV and HCV screening, which may have an important impact upon TB management and treatment.