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[Comparative study of the detection of plasma folate with microbial assay and radioimmunoassay].

OBJECTIVE: To compare two methods (microbial assay and radioimmunoassay) for measuring plasma folate concentrations, and to examine the relationship between plasma folate levels, and alcohol consumption, tobacco use and body mass index, and the risk of hyperhomocysteinemia in China. METHODS: We used a microtiter plate microbial assay and a radioimmunoassay to measure the folate concentration in 88 plasma samples. After comparing the results of these two methods and fitting a regression line, we examined the geographical, seasonal, and gender differences in folate concentration of plasma collected from 2,422 adults in south and north areas in China, and evaluated the association of plasma folate concentration, with alcohol consumption, cigarette smoking, and body mass index, and with the risk of hyperhomocysteinemia, using the data from the two assays. RESULTS: The data from the two assays had a linear relationship (r=0.879, P=0.000); the regression was Y=0.683X+0.308 (where X and Y were nature logarithmic transformations of plasma folate by microbial assay and radioimmunoassay, respectively); however, the mean plasma folate levels by microbial assay were much higher than those obtained by radioimmunoassay. Both data sets showed similar plasma folate distributions among Chinese adults, associations with other risk factors, and the risk of hyperhomocysteinemia. We estimated that 19.9% of the Southerners and 67.1% of the Northerners had plasma folate concentrations by radioimmunoassay lower than the 6.8 nmol/L used to define plasma folate deficiency. CONCLUSION: There is a linear relationship between plasma folate levels determined by microbial assay and radioimmunoassay, but because of the different levels obtained in the two assays, it is difficult to use the microbial assay results to evaluate folate status at this time. The use of 10.5 nmol/L as a cut-off for plasma folate deficiency by microbial assay needs further study.

Adult↗

[Clinico-pathological collations in borreliosis].

Borreliosis is an infectious disease caused by spirochetal microorganisms Borrelia burgdorferi transmitted by ticks. Due to versatile clinical symptomatology and many pathogenetic aspects not explained yet, diagnosis of this disease is often very difficult. Borreliosis may affect various human organs and systems such as movement system, nervous system (central and peripheral), skin, heart and vessels. One of the diagnostic methods for detection of microbial organisms in tissues is the histochemical staining by Warthin-Starry. Very important problem connected with this illness is the relationship between borrelial antigens and autoimmunity.

Humans↗

[Establishment of in vitro cultivation of Giardia canis trophozoites infected with Giardia canis virus].

OBJECTIVE: To cultivate a Giardia canis isolate with G. canis virus (GCV). METHODS: Five-day-old Meriones unguiculatus was infected with the cysts of G. canis isolated from dogs in Changchun and purified by sucrose density gradient centrifugation-G1 acid funnel filtration method. Trophozoites were isolated aseptically from the duodenum of the infected rodent after 8 days, then transferred to modified TYI-S-33 medium and cultivated at 37 degrees C. The trophozoites were centrifuged with 3,000 x g, 15 min after liquid nitrogen freeze-thawing three times and the supernatant stained negatively by phosphotungstic acid was observed with transmission electron microscope. RESULTS: G. canis trophozoites which adapted gradually to the environment and grew a cellular monolayer after 14 days were examined by freezing and thawing experiment, purity quotient, stability, biology characteristics and microbial contamination detection. The results demonstrated that a stable G. canis trophozoite cell isolate was established. G. canis virus with icosahedron spherical shape and 36 nm in diameter was observed by electron microscope. CONCLUSION: In vitro cultivation of G. canis trophozoites with GCV is established.

Animals↗

[Effects of Arbuscular Mycorrhizae on growth and essential oil of Atractylodes lancea].

OBJECTIVE: To study the effects of Arbuscular Mycorrhizae on cultivated Atractylodes lancea. METHOD: Pot experiment of A. lancea, with (code as AM) or without (code as CK) Glomus mosseae (GM) was conducted 5 times respectively, then the biomass, essential oil, and soil nutrition, soil organism, soil microbial were detected after A. lancea were harvested. RESULT: (1) Mycorrhizal dependency of A. lancea was 245%, and height of individuals, numbers of leaves, leaf area, biomass of A. lancea were all higher in AM than in CK (P < 0.05). (2) GC-MS analysis with cluster analysis and principal components analysis showed that there were no differences in essential oil of A. lancea between AM and CK. (T3) Total N, available N, available P and available K in AM soil were all lower than in CK soil. (4) GC-MS analysis showed organic matters changed differenly in AM soil and CK soil, components 5,6 in AM soil were higher than that in CK soil, but component 9, 10, 11 were lower in AM soil than that in CK soil. (5) Biolog detect showed AWCD of AM soil microbe were higher than that of CK soil throughout the incubation, and AWCD of the former was 0.66, and the later was 0.46 after 192 h incubation. and t-test showed, Shannon seven indices and McIntosh'seven indices were same both at 72 h and 192 h, and diversity indices of Shannon and McIntosh were also same at 72 h, but AM soil microbe were higher than CK soil microbe at 168 h (P < 0.05). CONCLUSION: AM could promote nutrition uptake, improve the function diversity and activity of microbe in rhizosphere of A. lancea, influence the composition of the organic matter, that lead the growth of A. lancea, but not to the quality.

Atractylodes↗

Microbial transformation of hexachlorocyclohexane.

Some microorganisms are able to accumulate and to metabolize the pesticide lindane (gamma-hexachlorocyclohexane, gamma-HCH) and other HCH-isomers. Microbial activities, detected metabolites, transformation rates, concentrating factors are reviewed. The results are discussed in respect to use microbiological processes for purification of contaminated water and soil and for recycling of waste HCH-isomers.

Aerobiosis↗

Fluorogenic detection of atypical coliforms from water samples.

We examined the effectiveness of fluorogen in detecting bacterial enzymes in atypical or injured coliform strains in environmental water samples. 4-methylumbelliferyl-beta-D-galactoside and 4-methylumbelliferyl-beta-D-glucuronide, substrates for beta-galactosidase and beta-glucuronidase respectively, were used as markers for total and faecal coliform bacteria and it was confirmed that fluorogenic assays have a greater sensitivity than reference methods. It was also observed that adding MU-conjugates (50 micrograms/ml) to low selective media for membrane filtration, besides shortening test times, reduces false negative results when detecting sanitary microbial indicators of water pollution.

Bacteria↗

[New developments in bedside diagnosis].

The time required to isolate microorganisms, identify them, interpret results, and provide guidance for therapy is inordinate for urgent decisions in primary care. The emphasis for a more rapid diagnosis bedside or in the office, respectively, is caused apart from medical reasons by economic restrictions necessitating earlier diagnosis, more immediate and appropriate treatment, shorter patient hospital stays, and allowing the patient to resume normal productivity earlier. After describing some general criteria with regard to the design of tests for bed-side diagnosis we discuss well known and novel procedures: microscopy of clinical specimens, microbial antigen detection, and nucleic acid hybridization. In concluding remarks we underline the necessity of careful clinical studies concerning sensitivity, specificity, and reliability as well as quality control measures for every novel assay before and during introduction in clinical and office practice.

Bacteria↗

Legionnaires' disease: ultrastructural appearance of the agent in a lung biopsy specimen.

During the Legionnaires' disease epidemic that occurred in Philadelphia in 1976, we performed a transbronchial lung biopsy on a patient who suffered from Legionnaires' disease that was confirmed by serology. The biopsy was performed in an attempt to detect a causal agent for the patient's pneumonia at a time when the cause of the epidemic was a mystery. We detected pleomorphic microbial structures that were both intracellular and extracellular in location. By electron microscopy, they ranged in size from 350 nm to 2.5 mu. Some displayed cell walls and fine cytoplasmic granules that resembled ribosomes. Our findings illustrate the appearance of the Legionnaires' disease agent in acutely infected, antibiotic-treated human lung obtained from biopsy specimens.

Biopsy↗

Aspergillus antigen detection in bronchoalveolar lavage fluid from patients with invasive aspergillosis and aspergillomas.

Improved diagnostic techniques have been needed for pulmonary aspergillosis, a common opportunistic fungal infection with a high mortality rate. Radioimmunoassay was used in this study to detect Aspergillus antigen in bronchoalveolar lavage fluid. In four patients with invasive aspergillosis or aspergillomas, Aspergillus antigen was detected in bronchoalveolar lavage fluid. In two patients, results of fungal cultures were negative or delayed. The specificity of antigen detection in bronchoalveolar lavage fluid was 91 percent in 35 control patients with a variety of pulmonary disorders. The technique or radioimmunoassay detection of microbial antigen in bronchoalveolar lavage fluid appears promising for the diagnosis of aspergillosis.

Aged↗

Microbiological evaluation of women with bad obstetrics history.

In the present study a total of 300 pregnant women were evaluated, 200 women with bad obstetrics history (BOH) and 100 clinically normal women. Cervical culture studies, as well as serological evaluation was carried out in each woman. It was noted that among the various microbial agents detected, the presence of genital mycoplasmas, chlamydia, Toxoplasma gondii and cytomegalovirus was significant. Foetal outcome could be noted in some of the BOH patients. Toxoplasmosis was associated with complete abortion (38%), stillbirths (6%), premature delivery (16%) and congenital anomalies (6%). Cytomegalovirus infection was associated with complete abortion (41.66%), preterm delivery (33.33%) and congenital anomalies (8.33%). Ureaplasma infection in BOH patients resulted in preterm delivery with premature rupture of membranes in 45 per cent of women and complete abortion in 35 per cent.

Case-Control Studies↗

[Current state and problems in the microbiology laboratory--organizing the laboratory].

The microbiological laboratory in the hospital has many roles including the rapid and precise identification of pathogenic bacteria in specimens, their antimicrobial susceptibility tests, microbial antigen detection using immunological methods and DNA hybridization methods, surveillance of bacterial milieu of the hospital environment, monitoring quality control of microbiological methods, to educate microbiological skills of staffs, the economical management of laboratory and so on. The last two issues are our major concerns. Improvement of the microbiological skills of the staff is most important in the laboratory, but is a time-consuming. From our experience, a technical expert, intermediately skilled technicians and beginners should always work in the laboratory, together. On the other hand, the economical management of the microbiological laboratory is also another concern. Although the mechanization of procedures comes to mind, it is hard to improve the economical conditions in the laboratory. Because of the control of the machines is not completely automatic and requires the knowledges and decisions of technical experts, the work force and running cost can not be reduced. As the technical expert can economize in the use of media or tests for identification of pathogens, good training of beginners into skillful successors, is important.

Humans↗

[The possible use of a ready-to-us film system for microbiological control in drug preparation].

Aim of this work was to verify the possibility of using a ready-to-use plate-count method to detect the microbial and fungal contamination on pharmaceutical products. The system consists of a flexible polypropylene film, supporting a suitable dehydrated medium, a second support containing guar, and an indicator on the internal surface. 33 raw materials, 11 natural origin materials, 20 medicinal product of official formula and 18 homeopathic products were analysed. As reference we chose the Italian Pharmacopoeia method. The two methods were comparable, showing no statistical differences, but for one case. This method, if our date will be further confirmed, could be used by the pharmacies and by the homeopathic industries.

Drug Compounding↗

[Applying an ultramicroanalytical system to the diagnosis of infectious diseases].

The multiple applications and advantages of the ultramicro analytical system in the serodiagnosis of different diseases produced by viruses, bacteria, and parasites are briefly reported. Also, perspectives in the use of this technology, not only for the diagnosis, but for the detection of microbial antigens are pointed out.

Communicable Diseases↗

[Laboratory diagnosis of Lyme borreliosis].

Some direct methods that can be used for the diagnosis of Lyme borreliosis are the culture, direct visualization or the detection of microbial DNA using polymerase chain reactions, in body tissues or fluids. Unfortunately, all these methods have a low sensitivity. There is a wide assortment of tests and antigens for indirect diagnosis, and the most recommended are ELISA tests and Western blot. The main inconvenient of these tests are the existence of shared serologic reactions, the variability of immune response and the difficult interpretation of results. Therefore, we propose the following guidelines for the diagnosis of Lyme borreliosis: For sero-epidemiological studies and to diagnose infection, antibodies should be determined in subjects with a compatible clinical picture, using an ELISA test that must be positive in at least two separate samples. All positive ELISA results should be confirmed with Western blot analysis, that must be interpreted using established criteria. Polymerase chain reactions should be used when they are available.

Borrelia burgdorferi Group↗