Parasites detected from diarrheal stool samples collected in Nepal

Intestinal parasites were investigated in 396 diarrheal stool samples collected from individuals aged 1 to 68 years (males: 239 and females: 157) in Nepal. Samples were collected at different medical centers located in Kathmandu and from two public schools in a village setting in Kathmandu Valley and outside, during October 1999 to January 2001. The stool samples were mixed with 2% dichromate solution and transported to Japan for investigations. Parasites were detected by employing the formal-ether sedimentation technique. Of a total of 396 fecal samples investigated, 193 (49%) were positive for some kind of parasite. Altogether, 15 species of parasites were detected. Giardia intestinalis topped the list of protozoa, whereas Trichuris trichiura was the most frequently detected among helminth parasites. Of the 193 positive samples, 109 (56%) had single parasite infections, whereas 84 (43%) had multiple infections with a maximum of five species. Of the total positive, 45 (23%) had both protozoa and helminths whereas 37 (19%) had only protozoa. Females (52%) and children (15 years and under) (52%) had a marginally higher prevalence compared with males (46%) and adults (45%), respectively (p > 0.05). Samples collected from two public schools in a village setting inside Kathmandu Valley and outside had a significantly higher positive rate compared with those observed in individuals visiting different medical centers in the city and suburban areas in Kathmandu (p < 0.05).     

IgM-capture ELISA of serum samples collected from Filipino dengue patients

Viral antigens for 4 dengue serotypes were produced in C6/36 Aedes albopictus cells. These were used as assay antigens for IgM-capture ELISA to detect IgM antibodies in sera of dengue patients from 3 hospitals in Metro Manila, Philippines. A total of 378 serum samples came from National Children's Hospital (NCH), San Lazaro Hospital (SLH), and St Luke's Medical Center (SLMC), from January to November 1995. Three hundred and four (304) out of 378 serum samples, or 80.42% showed positive IgM ELISA titer against at least one of the 4 assay antigens. Dengue type 4 (D4) antigen detected antibodies in 61.90% (234/378) of these serum samples, whereas type 1 (D1), type 3 (D3), and type 2 (D2) had detection rates of 60.05% (227/378), 50.79% (192/378) and 49.47% (187/378) respectively. Although the results show that both D1 and D4 are the most effective antigens in identifying dengue infections for this batch of samples, the use of a cocktail of antigens is still recommended. The results of this study are the basis for the IgM-capture ELISA protocol presently applied for the laboratory confirmation of dengue cases in the Philippines.     

Analysis of bronchoalveolar lavage samples collected from 30 patients with drug-induced pneumonitis

BackgroundDrug-induced pneumonitis is a disease encountered by pulmonologists in the clinical setting. The diagnosis generally considers the patient's clinical course and the results of peripheral blood tests, radiological examinations, and often bronchoscopic examinations. However, few studies have reported the association between radiological patterns such as ground-glass opacity (GGO) or consolidation, and bronchoalveolar lavage fluid (BALF) cell fractions. This study aimed to clarify this association.MethodsPatients with a Naranjo's score of probable or definite were enrolled, and all 30 patients were categorized under probable. Data such as patient background, blood examination results, radiological findings, and BALF cell fractions were retrospectively collected. The association between BALF cell fractions and other factors such as chest computed tomography (CT) findings was evaluated.ResultsThe most common radiological finding in patients with lymphocyte-dominant BALF was GGO, with only one patient exhibiting consolidation. However, patients with eosinophil-dominant BALF were more likely to have consolidation; only three cases showed crazy paving and one showed GGO. In addition, patients with a GGO-dominant pattern on CT had an increased lymphocyte fraction of 41.0%; those with a consolidation-dominant pattern showed a relatively high eosinophil fraction of 5.2%; and those with a crazy paving pattern showed elevated eosinophil and neutrophil fractions of 19.1% and 9.9%, respectively.ConclusionsIn this study, a remarkable difference in radiological findings was observed among different BALF patterns.     

Cyclospora spp. in herbs and water samples collected from markets and farms in Hanoi, Vietnam

Objective To determine the prevalence of Cyclospora spp. oocysts in herb and water samples as well as in fecal specimens of clinical cases of diarrhoea in Hanoi, Vietnam. Method From November 2004 to October 2005, water and herb samples collected from markets and farms in Hanoi were examined for the presence of Cyclospora spp. oocysts in concentrated sediments and washings using UV epifluorescence examination of a wet mount. In addition, hospital based surveillance studies were carried out using a structured questionnaire which focused on potential risk factors for cyclosporiasis. Stool specimens were collected from individuals with diarrhoea attending primary healthcare facilities and examined for Cyclospora spp. oocysts by modified acid fast smear and wet mount examination using both light and UV epifluorescence microscopy. Results Cyclospora spp. were found in 34/288 (11.8%) market water and herb samples, and in 24/287 (8.4%) farm samples. All varieties of herbs sold at the market and grown in farms were contaminated with Cyclospora spp. oocysts. A marked seasonal increase in Cyclospora spp. contamination was observed before the rainy season (39/288) from November to April compared to the rainy season (19/268) from May to October (χ² = 7.593, P = 0.006). However, Cyclospora spp. was not found in any stool samples collected in hospital‐based surveillance studies. Conclusions These results confirm the presence of Cyclospora spp. which varies seasonally in environmental samples (water and herbs collected from farms and markets) within the Hanoi metropolitan area.     

Seasonal distribution of enteropathogens detected from diarrheal stool and water samples collected in Kathmandu, Nepal

A total of 334 diarrheal fecal samples (from 210 males and 124 females) collected in Kathmandu, Nepal, were studied for various kinds of enteropathogens. Overall, 33% (111/334) fecal samples were positive for one or more enteropathogens. There was no difference in detection rates between males and females. Enteropathogen detection rates in summer, winter, spring, and autumn were 61% (40/66), 52% (45/87), 31% (25/81), and 25% (25/100), respectively. Altogether eight species of bacteria, three genera of viruses, and five species of protozoan parasites were detected with considerable seasonal variations. Among the bacterial isolates, enteropathogenic Escherichia coli topped the list followed by Vibrio sp. Only one sample had Shigella (S. sonnei). Rotavirus type A was the most frequently detected among the enteric viruses, followed by human enterovirus and human adenovirus, respectively. Among the enteric protozoan parasites, Giardia intestinalis was the most frequently detected followed by Cryptosporidium parvum. Detection of bacterial and protozoan pathogens showed a slightly high tendency in the summer season compared with that in the other seasons (p>0.05), whereas the detection of viruses was significantly high in the winter season (p<0.05). Of the total 57 water samples, 43 (75%) showed one or more bacterial species out of which 51% (22/43) were E. coli. Among the E. coli isolates, 68% were EPEC. Enterohemorrhagic E. coli (O157) was not detected.     

丙型肝炎病人系列血清中5'-NCR和C区基因变异研究

目的研究丙型肝炎病人的系列说清中ＨＣＶ ５'－ＮＣＲ及核心区基因变化。方法 ４例因献血感染ＨＣＶ病人的系列血标本,用特异性引物扩增５'－ＮＣＲ和核心区基因并进行测序。结果所有ＨＣＶ分离株基因序列均为１ｂ和２ａ型,５'－ＮＣＲ序列差异只与型别有关,与病人和不同时点采集的标本无关。核心区基因序列差异因病人和基因型别而异,未发现同一病人的同一基因型序列随时间而变化。结论５'－ＮＣＲ序列保守性大于核心区。型别是导致基因变化的主要原因。未发现该２区段基因的核苷酸改变与时间有关。     

The level of allergens in dust samples collected from selected schools in Shiraz,Iran and its asthma-risk implications

BackgroundBoth home and school are important places where children are exposed to various indoor allergens. This study aimed to identify the profile of indoor allergens in schools and its impact on asthma development.MethodsA total of 104 classrooms from 52 schools were selected for dust collection during the fall of 2017. The levels of indoor allergens including dust mite (Der f1, Der p1), cat (Fel d1), cockroach (Bla g1) and mouse (Mus m1) were measured by enzyme linked immunosorbent assay (ELISA). The diagnosis of asthma was made in all students of the selected classes by the allergist. The collected data were analyzed using SPSS version 21.0.ResultsOut of 2816 students in the selected classes, 180 students were involved with asthma. Students were mostly exposed to Bla g1 (83.1%), followed by Der f1 (51.5%), Mus m 1 (45.5%), Der p1 (8.9%) and Fel d1 (7.9%) in the dust collected from 101 classrooms. Although levels of all studied allergens in the settled dust of the classrooms were low, there was a relationship between Fel d1 in the classroom dust and development of asthma.ConclusionThis study showed considerable levels of cockroach allergens in schools. Exposure to cat allergen in our schools played an important role in asthma development; further school-based investigations require evaluating the role of classroom allergen on asthma development.     

Fluorescent treponemal antibody absorption (FTA-ABS) tests using blood samples collected on filter paper.

One hundred and eleven (111) blood specimens were collected from patients with known or suspected syphilis by venipuncture and on a small piece of No. 3 grade filter paper. Both specimens from each patient were tested by bluorescent treponemal antibody absorption (FTA-ABS) test. Of the 62 patients whose venipuncture sera were FTA-ABS positive, filter paper blood samples were also positive in 56 (90%) and negative in 6 (10%). Complete agreement was obtained on paired specimens from 49 patients who were FTA-ABS negative. These preliminary results suggest that finger stick blood samples, collected on filter paper, could be used for FTA-ABS testing of remote rural populations--such as in areas where yaws is endemic.     

贝氏柯克斯体实时荧光定量PCR方法的建立及对云南鼠标本检测

目的 建立贝氏柯克斯体荧光定量PCR方法并应用于云南省鼠标本的检测.方法 根据贝氏柯克斯体ISlllla基因设计引物和探针,以克隆的ISlllla基因片断(485 bp)作标准DNA模板,建立实时荧光定量PCR检测方法,并与普通巢式PCR方法进行比较.采用2种方法对云南玉溪自然界采集的鼠各类标本进行检测分析,计算2种方法的检出率.结果 实时荧光定量PCR标准曲线的循环阈值(cycle threshold,Ct)与模板拷贝数呈良好的线性关系(r=-0.999),重复性测试Ct变异系数(coefficient of variation,CV)为0.25%～3.98%,灵敏度为巢式PCR的10倍.以其他立克次体和常见病原菌共26种DNA为模板进行特异性检测,结果均为阴性.实时荧光定量PCR检测鼠血、肝脏、脾脏中的贝氏柯克斯体,阳性率分别为32.31%、61.29%、85.19%;巢式PCR法检测相应标本阳性率分别为27.69%、11.29%、74.07%;间接免疫荧光试验检测鼠血清中贝氏柯克斯体IgG抗体阳性率为26.15%.结论 本实验建立的实时荧光定量PCR比巢式PCR敏感,且具有良好的特异性,可用于人群和动物贝氏柯克斯体感染监测及临床标本的检测.     

Short report: Diagnosis of human fascioliasis: detection of anti-cathepsin L antibodies in blood samples collected on filter paper.

We have developed an ELISA for the diagnosis of human fascioliasis based on the detection of IgG4 antibodies to Fasciola hepatica cathepsin LI cysteine protease. Use of this assay in the Bolivian Altiplano, a region with a high prevalence of the disease, was hampered by the reluctance of the indigenous population to provide blood. To overcome this problem, we have investigated the method of collecting small quantities of blood from the finger onto filter paper, followed by the elution of antibodies for use in the diagnostic assay. Serum samples and blood samples collected onto filter paper were obtained from 57 individuals living in the village of Cutusuma in 1987 and from 11 individuals in Chijipata in 1996. Analysis of the IgG4-ELISA results revealed that there is highly significant linear relationship (P < 0.001) between the two methods of sampling. Most importantly, a reliable diagnosis was made with the blood-filter samples from Cutusuma, which had been stored for 10 years at 40 degrees C. While some deterioration of the blood-filter samples from Cutusuma had occurred over the 10-year storage period, no deterioration occurred with the Chijipata samples, which were stored for one year. Therefore, the method of collecting blood onto filter paper should prove useful for large-scale epidemiologic studies on human fascioliasis in the Bolivian Altiplano and in other regions where this disease is prevalent.     

Chemical analysis and antioxidant content of various propolis samples collected from different regions and their impact on antimicrobial activities

Objective: To assess the antioxidant content, antimicrobial and antioxidant activities of various propolis samples. Methods: Seven propolis samples were collected from different locations in Morocco, which are characterized by different plant predominant vegetations. The resin, wax and balsam of hydroalcoholic extract of propolis content were identified, and the antioxidant content was analyzed with the use of HPLC and colorimetric methods. The antioxidant activity was assessed by DPPH, ABTS.~+ and ferric reducing power assays. The antimicrobial activity was assessed against bacterial species, including methicillin resistant Staphylococcus aureus and Candida albicans, and expressed as the minimal inhibitory concentration. Results: The propolis samples showed significant variations in the chemical composition and in the antioxidant or antimicrobial activities even when the samples were collected from the same location. Propolis with high resin and low wax content had high level of antioxidant compounds, and strong antioxidant and antimicrobial activities. Gram-positive bacteria, especially, methicillin resistant Staphylococcus aureus were more sensitive to all propolis samples than Gram-negative bacteria and Candida albicans. Conclusions: The chemical composition and the antioxidant and antimicrobial activities of various propolis samples are different and rely on the geographic and plant origin of propolis collection. Propolis samples with low wax and high resin content might be more suitable to be used in future preclinical or clinical investigations.     

Early prediction of treatment outcome in acute myeloid leukemia by measurement of WT1 transcript levels in peripheral blood samples collected after chemotherapy

The Wilms' tumor gene WT1 is a reliable marker for minimal residual disease assessment in acute leukemia patients. The study was designed to demonstrate the potential use of WT1 to establish quality of remission in acute leukemia patients for early identification of patients at high risk of relapse. A prospective study based on a quantitative Real-Time PCR (TaqMan) assay in 562 peripheral blood samples collected from 82 acute leukemia patients at diagnosis and during follow-up was established. The evaluation of WT1 in peripheral blood samples after induction chemotherapy can distinguish the continuous complete remission patients from those who obtain only an "apparent" complete remission and who could relapse within a few months. WT1 helps identify patients at high risk of relapse soon after induction chemotherapy allowing post-induction therapy in high risk patients to be intensified.     

Neutralizing antibodies against two HIV-1 strains in consecutively collected serum samples: cross neutralization and association to HIV-1 related disease.

97 sera collected during a 10-year period from 10 HIV-1 infected individuals were tested for neutralizing capacity against a virus isolate FICPH-22 obtained from a Danish AIDS patient, and the laboratory strain HTLV-IIIB. Three patterns of serum neutralizing activity were demonstrated: (a) patients developing high neutralizing activity against both HIV strains; (b) patients developing high neutralizing activity against the Danish virus isolate; and (c) patients developing only low titers of neutralizing antibodies (NA) against both HIV strains. The HTLV-IIIB strain was less sensitive to serum neutralization than the FICPH-22 isolate and the appearance of NA against HTLV-IIIB was typically lacking several years behind that against FICPH-22 indicating a broadening of the NA response over time. No difference in clinical outcome was observed comparing patients reaching high titers of NA and patients with low titers. Development of AIDS among patients reaching high titers of NA was preceded by a decline in NA titers, indicating an association of high titers of NA with the healthy carrier state and of declining or low titers of NA with disease progression. The majority of the neutralizing activity was mediated by IgG, but some neutralizing activity was demonstrated in the IgG depleted serum, indicating the presence of additional neutralizing substances in serum.     

Strengths and weaknesses of established indirect models to detect recombinant human erythropoietin abuse on blood samples collected 48-hr post administration

We studied indirect detection models of erythropoietin abuse (EPO) on blood samples collected 48-hr after administration of the drug during 6 weeks of recombinant human erythropoietin (rHuEPO) treatment. Although the efficiency of OFF-models was preserved, we found a loss of sensitivity of ON-models. This study also revealed an increased percentage of stomatocytes in athletes receiving rHuEPO