High-performance liquid chromatography and Enzyme-Linked Immunosorbent Assay techniques for detection and quantification of aflatoxin B1 in feed samples: a comparative study.

ObjectiveComparison was done between high-performance liquid chromatography (HPLC) and a competitive enzyme-linked immunosorbent assay (ELISA) for detection and quantification of aflatoxin B1 (AFB1) in feed samples. The two procedures were standardized and validated before the actual experiment. Five concentrations (0, 5, 10, 20 and 30 ppb) of feed samples were used for both methods. For the HPLC technique, the samples were extracted in acetonitrile/water (90/10) solution, cleaned-up using solid phase extraction (SPE) column, and derivatized by water/trifluoroacetic acid/glacial acetic acid (35/10/5) solution before instrument analysis. The samples were extracted in 70% methanol for the ELISA technique.ResultsThe two tests showed very strong linearity with correlation coefficient value of > 0.99 using standard solutions. The mean recovery rate was 92.42% (with relative standard deviation (RSD) of 5.97) and 75.64% (RSD = 34.88) for HPLC and ELISA, respectively. There was no statistically significant difference in recovery rate between the two methods. There was a positive correlation (r = 0.84) between them which indicated that the two techniques can be used to detect and quantify aflatoxin B1 in feed samples. However, there were variations among replicates for the ELISA method, which shows that this method is more applicable for screening purposes

Bacteriology and antibiogram of pathogens from wound infections at Dessie Laboratory, North East Ethiopia

Wound infections result in sepsis, limb loss, long hospital stays, higher costs, and are responsible for significant human mortality and morbidity worldwide. The aim of this study was to investigate the profile of pathogens cultured from wound infection and determine the antimicrobial susceptibility patterns.  A retrospective analysis of bacterial pathogens and their antimicrobial susceptibility was done on wound swab samples that have been cultured at Dessie Regional Laboratory from 2003 to 2010. Antimicrobial susceptibility tests were done using disc diffusion technique as per the standard of Kirby-Bauer method. The mean age of male and female patients was 31.2 and 29.8 years, respectively with male to female ratio of 1:1.6. Out of 599 wound swab samples analyzed, 422 (70.5%) were culture positive.  A total of 500 bacteria from 422 positive cases were identified. Seventy eight (18.5%) of the culture had double infections. Staphylococcus aureus was the most frequently isolated pathogen which accounted for 208 (41.6%) of isolates followed by Pseudomonas spp. 92 (18.4%), E. coli 82 (16.4%), Proteus spp. 55 (11.0%), Enterobacter spp. 21 (4.2%), and Citrobacter spp. 21 (4.2%), Klebsiella spp. 12 (2.4%) and Coagulate negative staphylococcus 9 (1.8%). Amoxicillin had the highest resistance rate 78.9%, followed by tetracycline 76.1% and erythromycin (63.9%). The sensitivity rates of norfloxacin, ciprofloxacin and gentamicin were 95.1%, 91.8% and 85%, respectively.  The overall multiple antimicrobial resistances rate was 65.2% and only 13% of the isolates were sensitive to all antimicrobial agents tested. The most frequently isolated bacteria were sensitive to ciprofloxacin, gentamicin, cloxacillin and norfloxacin. These antimicrobials are considered as appropriate antimicrobials for empirical treatment of wound infections. Periodic surveillance of aetiology and drug susceptibility both in the community and hospital settings is recommended.&nbsp

Na-K-Cl Cotransporter-1 as a Regulator of Manganese-induced Astrocyte Swelling

Astrocyte swelling leads to brain edema, intracranial pressure, brain herniation and acute liver failure (fulminant hepatic failure) which is the major cause of death in this condition. Manganese has been strongly implicated as an important factor in astrocyte swelling. Manganese in excess is neurotoxic and causes a CNS disorder that resembles  Parkinson¡¦s disease (manganism). Manganese highly accumulates in astrocytes, which renders these cells more vulnerable to its toxicity. In addition to manganism, increased brain levels of manganese have been found in hepatic encephalopathy. Manganese is known to cause cellswelling in cultured astrocytes, although the means by which this occurs has not been fully elucidated. A disturbance in one or more of these systems may result in loss of ion homeostasis and cell swelling. In particular, activation of the Na-K-Cl cotransporter-1 (NKCC1) has been shown to be involved in cell swelling in several neurological disorders.We therefore examined the effect of manganese on NKCC activity and its potential role in the swelling of astrocytes. Cultured astrocytes were exposed to manganese (50 µM), and NKCC activity was measured. Manganese increased NKCC activity at 24 h. Inhibition of this  activity by bumetanide diminished manganese-induced astrocyte swelling.  Manganese (Mn) also increased total as well as phosphorylated NKCC1. These results suggest that activation of NKCC1 is an important factor in the mediation of astrocyte swelling by manganese and that such activation appears to be mediated by NKCC1 abundance

Patient satisfaction with community pharmacist-led anticoagulation management services and its relationship with patient characteristics in New Zealand

BACKGROUND: Community pharmacist-led anticoagulation management service (CPAMS) offers international normalised ratio point-of-care testing of warfarin in a community pharmacy setting. It has now expanded with 7,344 patients enrolled in the service across 164 pharmacies in New Zealand. The clinical benefit of CPAMS has been shown to be superior, but patient satisfaction with the service has not been fully explored. OBJECTIVE: To develop a questionnaire to assess patient satisfaction with CPAMS and evaluate its psychometric properties. Additionally, to determine the level of patient satisfaction with CPAMS and identify determinants of satisfaction with CPAMS. Settings 1071 patients enrolled in CPAMS across New Zealand invited to take part in the study. MAIN OUTCOME MEASURE: Satisfaction with CPAMS service. METHODS: Adult patients taking warfarin and currently enrolled in CPAMS were recruited through the national international normalised ratio online system and invited to complete a 36-item survey assessing satisfaction with CPAMS. To identify the most important dimensions of patient satisfaction, exploratory factor analysis was used. Multivariate linear regression models were used to examine the effect of independent variables on patient satisfaction. RESULTS: A total of 305 patients completed the survey. The mean overall satisfaction score was 94.5% ± 13.1 out of maximum possible points. Five dimensions of patient satisfaction were identified by factor analysis: patient-centred communication, confidence in pharmacist competence, patient-pharmacist relationship, confidence in CPAMS, and pharmacy environment. Being older and more frequent visits to a pharmacy were positively associated with patient satisfaction. Living more than 1 km away from a pharmacy, and ‘poor’ self-perceived health status were negative predictors of patient satisfaction. Being Māori or of other ethnic minority was also associated with lower satisfaction scores, exploratory analysis suggests patient-pharmacist relationship is an important driver of these differences. CONCLUSIONS: The high level of patient satisfaction further supports the effectiveness of CPAMS as a delivery model. Patient satisfaction is affected by age, frequency of pharmacy visits, ethnicity, travel distance to pharmacy, and perceived health status. Policy makers and practitioners should consider the characteristics of patients with low levels of satisfaction to improve and enhance CPAMS engagement

Responses of tropical maize landraces to damage by Chilo partellus stem borer

The potential to manage insect pests using host-plant resistance exists, but has not been exploited adequately. The objective of this study was to determine the resistance of 75 tropical maize landraces through artificial infestation with Chilo partellus Swinhoe. The trial was laid in alpha-lattice design and each seedling was infested with five neonates three weeks after planting, over two seasons in 2009 and 2010. The number of exit holes, tunnel length, ear diameter, ear length, plant height, stem diameter, stem lodging and grain yield were measured and a selection index computed. GUAT 1050 was the most resistant with an index of 0.56, while BRAZ 2179 was the most susceptible with an index of 1.66. Ear characteristics were negatively correlated with damage parameters. The principal component biplot suggested that exit holes, cumulative tunnel length, leaf damage, cob diameter, stem lodging, selection index, ear and plant height contributed 71.2% of the variation in resistance. The mean number of exit holes and tunnel length for resistant landraces and resistant hybrid checks were similar; at 5.5 and 2.48 cm, respectively. The identified resistant landraces (GUAT 1050, GUAT 280, GUAT 1093, GUAT 1082, GUAT 1014, CHIS 114, and GUAN 34) could be used to develop C. partellus stem borer-resistant maize genotypes.Key words: Chilo partellus, ear length, exit holes, stem borer resistance, tunnel length

Cryptosporidium in Calves, Lambs and Kids at Haramaya, eastern Ethiopia

A cross sectional study was carried out from November 2010 to April 2011 to determine the prevalence of Cryptosporidium in calves, lambs and kids and to assess potential risk factors in Haramaya, eastern Ethiopia. Hence, fecal samples from a total of 237 study animals (133 calves, 63 lambs and 41 kids) were tested with Sheather’s flotation technique and Modified Ziehl-Neelsen Staining. Accordingly, the overall prevalence was found to be 23.6% (56/237) with the prevalence of 27.8% (37/133) in calves, 22.2% (14/63) in lambs and 12.2% (5/41) in kids. The prevalence of Cryptosporidium significantly (p = 0.047) varied with species of study animals being three times higher likelihood of occurrence in calves compared to its chance of occurrence in kids (OR = 2.8, 95%CI = 1.0, 7.6). No significant difference in prevalence was observed between kids and lambs. The occurrence of the disease was also assessed among the species of animals by taking into consideration different demographic characteristic of the animals including breed, age and sex. For caprine, the prevalence was higher in the cross breed animals (28.6%) followed by the breed of Hararge highland (25%) with no report of the disease in Boer and Ogaden breeds. In ovine, no association was observed between the infection occurrence and the potential risk factors. Analysis of the risk factors in bovine revealed a significant (p = 0.044) difference in disease occurrence among age groups with almost three times more likelihood of the disease occurrence in calves under 3 months than older age category(OR = 2.9, 95%CI = 1.0, 8.2). In conclusion, this study demonstrated the importance of Cryptosporidium in young ruminants with a higher prevalence among calves than lambs and kids.Key words: Cryptosporidium; Prevalence; Risk Factors; Calves, Lambs, Kids, Haramaya, Ethiopi

Clinical performance of methylation as a biomarker for cervical carcinoma in situ and cancer diagnosis: A worldwide study.

The shift towards primary human papillomavirus (HPV)-based screening has necessitated the search for a secondary triage test that provides sufficient sensitivity to detect high-grade cervical intraepithelial neoplasia (CIN) and cancer, but also brings an improved specificity to avoid unnecessary clinical work and colposcopy referrals. We evaluated the performance of the previously described DNA-methylation test (S5) in detecting CIN3 and cancers from diverse geographic settings in high-, medium- and low-income countries, using the cut-off of 0.80 and exploratory cut-offs of 2.62 and 3.70. Assays were performed using exfoliated cervical specimens (n = 808) and formalin-fixed biopsies (n = 166) from women diagnosed with cytology-negative results (n = 220), CIN3 (n = 204) and cancer stages I (n = 245), II (n = 249), III (n = 28) and IV (n = 22). Methylation increased proportionally with disease severity (Cuzick test for trend, P < .0001). S5 accurately separated women with negative-histology from CIN3 or cancer (P < .0001). At the 0.80 cut-off, 543/544 cancers were correctly identified as S5 positive (99.81%). At cut-off 3.70, S5 showed a sensitivity of 95.77% with improved specificity. The S5 odds ratios of women negative for cervical disease vs CIN3+ were significantly higher than for HPV16/18 genotyping at all cut-offs (all P < .0001). At S5 cut-off 0.80, 96.15% of consistently high-risk human papillomavirus (hrHPV)-negative cancers (tested with multiple hrHPV-genotyping assay) were positive by S5. These cancers may have been missed in current primary hrHPV-screening programmes. The S5 test can accurately detect CIN3 and malignancy irrespective of geographic context and setting. The test can be used as a screening and triage tool. Adjustment of the S5 cut-off can be performed considering the relative importance given to sensitivity vs specificity

A randomized controlled trial of the effects of a prudent diet on cardiovascular risk factors, gene expression, and DNA methylation - the Diet and Genetic Intervention (DIGEST) Pilot study

Background Risk of cardiovascular disease (CVD) can be increased by single-nucleotide polymorphisms (SNPs) in the 9p21 region of the genome. However, observational studies have shown that the deleterious effect of 9p21 SNPs on CVD might be offset by consuming a diet rich in fresh fruits and vegetables. This association may be driven by diet-influenced modifications in epigenetic and gene expression profiles. In this pilot study, we aimed to: i. test the feasibility of provision of a ‘Prudent’ and ‘Western’ diet outside of a specialized clinic, ii. assess the impact of each diet on cardiovascular risk factors. Methods A single centre, parallel two-arm, pilot randomized controlled trial (RCT) with food provision was conducted in a university teaching hospital outpatient clinic (McMaster university, Hamilton, ON, Canada). The aim was to recruit 80 participants, which allowed for a 10 % dropout. The actual study consisted of 84 apparently healthy participants (69 % women, 18 to 77 years) at low cardiovascular risk. Participants were randomly assigned to follow one of two weight-maintaining diets: ‘Prudent’ or ‘Western’ for 2-weeks. The Prudent diet provided 92 % of provided food consumed). The Prudent diet was 48 % more palatable than the Western diet (P < 0.05). Participants receiving the Prudent diet showed a trend toward reduced systolic (-4 mmHg; P = 0.10) and diastolic (-3 mmHg; P = 0.07) blood pressure, and total cholesterol (-0.24 mmol/L; P = 0.08), compared to individuals receiving the Western diet. Data collection from all randomized participants was completed within 18 months. Conclusions Recruitment, and retention of apparently healthy, normotensive adults into a feeding study for a 2-week duration is feasible outside of specialized dietary clinic, and modest diet-related changes in biomarkers begin to appear after two weeks

Prevalence of Trachoma in Benishangul Gumuz Region, Ethiopia: Results of Seven Population-Based Surveys from the Global Trachoma Mapping Project.

PURPOSE: Trachoma is a major cause of blindness in Ethiopia, and targeted for elimination as a public health problem by the year 2020. Prevalence data are needed to plan interventions. We set out to estimate the prevalence of trachoma in each evaluation unit of grouped districts ("woredas") in Benishangul Gumuz region, Ethiopia. METHODS: We conducted seven cross-sectional community-based surveys, covering 20 woredas, between December 2013 and January 2014, as part of the Global Trachoma Mapping Project (GTMP). The standardized GTMP training package and methodologies were used. RESULTS: A total of 5828 households and 21,919 individuals were enumerated in the surveys. 19,583 people (89.3%) were present when survey teams visited. A total of 19,530 (99.7%) consented to examination, 11,063 (56.6%) of whom were female. The region-wide age- and sex-adjusted trichiasis prevalence in adults aged ≥15 years was 1.3%. Two evaluation units covering four woredas (Pawe, Mandura, Bulen and Dibate) with a combined rural population of 166,959 require implementation of the A, F and E components of the SAFE strategy (surgery, antibiotics, facial cleanliness and environmental improvement) for at least three years before re-survey, and intervention planning should begin for these woredas as soon as possible. CONCLUSION: Both active trachoma and trichiasis are public health problems in Benishangul Gumuz, which needs implementation of the full SAFE strategy