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Biomedical subjects

S Schaaf

Publications and source records attributed to S Schaaf.

11 recordsLinked to original sources
PubMed2001

Detection of Cryptosporidium oocysts in soil samples by enzyme-linked immunoassay.

An ELISA protocol was adapted for detection of Cryptosporidium parvum oocysts in soil samples and the limit of detection of the test was determined. A modified indirect antigen capture ELISA protocol was developed using monoclonal antibodies against the oocyst outer wall. The accuracy of the ELISA was compared to spiked soil samples and measured in terms of sensitivity and specificity of the test. The performance of the ELISA was evaluated in field soil samples and measured using the kappa-statistics. Similarly, the performance of the ELISA was compared to the concentration flotation method, to a modified concentration flotation method and to a commercial ELISA (ProSpecT) in field fecal and soil samples. The limit of detection of the test was selected to be 10,000 oocysts/g. At this limit of detection, the ELISA had a sensitivity of 95% and specificity of 100%. The agreement between the ELISA and the modified flotation-concentration method in detecting Cryptosporidium oocysts in soil samples was 32% (kappa=0.32). The ELISA had the same relative sensitivity (82%) in comparison to both the flotation and ProSpecT in determining Cryptosporidium-infection status of an animal. The kappa-statistics was 0.26 for both tests. The developed ELISA proved to be a valuable diagnostic test for detecting oocysts in soil samples and has a potential application in determining the infection status of animals.

Animals↗
PubMed1999

Risk factors associated with Cryptosporidium parvum infection in dairy cattle in southeastern New York State.

An observational analytical epidemiologic study was carried out to identify factors associated with the risk of infection with Cryptosporidium parvum in dairy herds in southeastern New York state. A random sample of 2943 cattle on 109 farms was selected from the target population. Fecal samples were collected from animals in three different age groups and examined for the presence of C. parvum using a quantitative centrifugation concentration flotation method. Data on intrinsic, preweaning, postweaning, maternity, and general management factors were collected and evaluated for their association with the risk of infection with C. parvum. Indices for each of these categories of management were developed from factors significantly associated with the risk of infection with C. parvum. Significant factors were identified using the logistic regression statistical technique. A final analysis, including the indices, age, and season, was performed to identify factors significantly associated with the risk of infection with C. parvum while simultaneously controlling for the effect of other factors. The farm effect was evaluated using a mixed effect model. Preweaning factors found to be significantly associated with a decreased risk of infection were: use of ventilation in calf rearing areas, daily addition of bedding, feeding of milk replacer, daily disposal and cleaning of bedding, and use of antibiotics. Postweaning factors such as moving of the animals after weaning, cleaning of soiled bedding, and use of antibiotics and ionophores as preventive measures were significantly associated with the decreased risk of an infection with C. parvum. Consideration of maternity management factors showed that winter housing of cows individually within 2 months of calving, use of fresh colostrum to feed calves, and having a concrete floor in the calving area were significantly associated with decreased risk of C. parvum infection. The total number of dairy cattle, total number of other species of agricultural animals on the farm, and the distance of the barn water source from the septic system were found to be significantly associated with increased risk of C. parvum infection. In the final analysis, the risk of infection with C. parvum was significantly decreased with an increased value of the maternity management index score. The general management significantly affected the risk of infection with C. parvum where the risk increased with the increase of the value of the index. The risk of infection significantly decreased with increase in the age of the animal.

Age Factors↗
PubMed1998

Survival of children in Cape Town known to be vertically infected with HIV-1.

OBJECTIVE: To determine the survival patterns of children in Cape Town known to be vertically infected with HIV. DESIGN: Retrospective record review of children diagnosed with symptomatic HIV infection during the period 1 December 1990-31 May 1995. SETTING: Hospitals in the Cape Town metropolitan area. PATIENTS: 193 children were known to be vertically HIV-infected. HIV diagnosis was based on the following criteria: two positive HIV enzyme-linked immunosorbent assays (ELISAs) in children older than 15 months and a positive ELISA together with a positive polymerase chain reaction (PCR) in younger children. The mothers of the children were known to be HIV-positive. On the basis of the presenting clinical findings children were assigned to a disease severity category (A, B or C) according to the Centers for Disease Control and Prevention (CDC)'s 1994 revised classification system for HIV infection in children. OUTCOME MEASURES: Survival was analysed according to the Kaplan-Meier method. Survival time was defined as the length of time between clinical diagnosis of HIV and death or last contact with the health services. Mortality risk in relation to specific variables at diagnosis such as age and clinical manifestations was determined by calculation of odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: The median age at diagnosis was 5 months; 72% of children were aged less than 1 year at diagnosis. According to the CDC clinical classification, 47 (24%) fell into category A, 111 (58%) into category B and 35 (18%) into category C. Of the 193 patients 85 (44%) were alive at the time of review, 65 (34%) had died and 43 (22%) were lost to follow-up. Risk of death was significantly associated with age less than 6 months (OR 4.7, CI 2.1-10.3) and severe disease, i.e. CDC category C (OR 2.7; CI 1.1-6.9) at time of diagnosis. The median survival for all the children from time of diagnosis was 32 months. Infants diagnosed before 6 months of age had significantly shorter median survival (10 months) compared with 36 months for those diagnosed at 7-12 months of age. For the children over the age of 12 months the cumulative proportion surviving at 48 months was 78%. Children with severe disease (category C) had a median survival of 21 months, significantly lower than that in category B (32 months). For the children in category A the cumulative proportion surviving at 48 months was 66%. CONCLUSION: The median survival of children with HIV was 32 months from time of diagnosis, and survival was influenced by age and disease severity.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Transgenic tobacco plants and their progeny derived by microprojectile bombardment of tobacco leaves.

Transgenic tobacco plants and progeny carrying coding sequences for neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) were recovered following microprojectile bombardment of tobacco leaves. Transgenic plants were regenerated from bombarded leaf pieces of tobacco cvs. 'Xanthi' and 'Ky 17' which were cultured in the presence of 100 or 200 micrograms/ml kanamycin for six to eight weeks. Among 160 putative transgenic plants from at least 16 independent transformation events 76% expressed NPTII, and 50% expressed GUS. Southern analysis of plants expressing either one or both of the enzymes indicated DNA in high molecular weight DNA in 8 of 9 independent transformants analyzed. Two independent transformants and their progeny were analyzed in detail. Analysis of progeny for quantitative enzyme levels of NPTII and GUS, and Southern analysis of parents and progeny clearly demonstrated that the genes were transmitted to progeny. One transformant demonstrated Mendelian ratios for seed germination on kanamycin-containing medium while the other transformant had non-Mendelian ratios. DNA analysis of progeny indicate complex integration of the plasmid DNA, and suggest that rearrangements of this DNA has occurred. These results are consistent with other methods of direct DNA uptake into cells, and verify that the microprojectile bombardment method is capable of DNA delivery into intact plant cells which can give rise to transgenic plants and progeny.

Blotting, Southern↗
PubMed1988

Transfer of foreign genes into intact maize cells with high-velocity microprojectiles.

This report describes a process for delivering foreign genes into maize cells that does not require the removal of cell walls and is capable of delivering DNA into embryogenic and nonembryogenic tissues. Plasmid harboring a chimeric chloramphenicol acetyltransferase (CAT) gene was adsorbed to the surface of microscopic tungsten particles (microprojectiles). These microprojectiles were then accelerated to velocities sufficient for penetrating the cell walls and membranes of maize cells in suspension culture. High levels of CAT activity were consistently observed after bombardment of cell cultures of the cultivar Black Mexican Sweet, which were comparable to CAT levels observed after electroporation of protoplasts. Measurable increases in CAT levels were also observed in two embryogenic cell lines after bombardment. Gene expression was observed only when an intron from the alcohol dehydrogenase 1 gene of maize was ligated between the 35S promoter and the CAT coding region. CAT activity was detected in cell cultures bombarded with microprojectiles with an average diameter of 1.2 mum, but not after bombardment with microprojectiles 0.6 or 2.4 mum in diameter. Bombarding the same sample several times was found to markedly enhance CAT activity. These results demonstrate that the particle bombardment process can be used to deliver foreign DNA into intact cells of maize. Because this process circumvents the difficulties associated with regenerating whole plants from protoplasts, the particle bombardment process may provide significant advantages over existing DNA delivery methods for the production of transgenic maize plants. In addition, the process should be of value for studying transient and stable gene expression within intact cells and tissues.

Journal Article↗
PubMed1984

Long-term development of a primary lung sarcoma, probably lymphangiosarcoma--a case report.

A small primary lung lesion had remained unchanged in size over a period of 8 years. Within the ensuing 2 years rapid tumor growth occurred. On operation the surgical specimen revealed a tumor mass of 87.2 cm3. Within 6 months after resection, the tumor occurred on the same side. Extensive tumor infiltration of the chest wall prevented any further surgery. The histo-morphologic picture of both lesions was consistent with a lymphangiosarcoma without evidence of high grade malignancy. The patient died of cardio-respiratory insufficiency due to extension of the tumor mass. Unfortunately no autopsy was performed. Tumor growth rates, macroscopic reconstruction and electron microscopic evaluations are presented. This case report again points out that size and long-time stability of solitary lung nodules are not necessarily reliable criteria for assessing prognosis.

Adult↗
PubMedDate not supplied

Transintestinal elimination of ciprofloxacin in humans--concomitant assessment of its metabolites in serum, ileum and colon.

Ciprofloxacin (200 mg) was infused to seven patients at the beginning of elective colorectal surgery. Thirty minutes after the end of infusion (i.e. 60 min after the start of the operation) ciprofloxacin reached concentrations of 1.60 mg/l in serum and of 3.42-6.07 mg/kg fresh weight in the ileum and colon. During the next 30 min (90 min after the start of operation) the concentration of ciprofloxacin in serum decreased to 86% of its initial level, but this decrease was less rapid than that observed in the ileal (to 56.8%) or colonic (to 74.8%) mucosa. Three metabolites could be identified (desethylen-, sulpho-, oxociprofloxacin). Initially, at 60 min the amount of these metabolites was about 15% of the total drug concentration in serum, but only 2-3% of that in the gut tissues. At 90 min the relative amount of metabolites was increased in serum as well as in the gut tissues. It is concluded that transintestinal elimination of ciprofloxacin is a general feature of the whole gut. Obviously, the elimination process is not due to degradation of ciprofloxacin within the gut wall.

Adult↗