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The male SHR rats infected with sialodacryoadenitis (SDA) virus did not show any abnormal mating activity. However, high mortality was seen in embryos of diseased dams after cage-mating with infected males on Day 4 to Day 11 postinoculation.
BACKGROUND: Chlamydia trachomatis is the most common bacterial sexually transmitted infection. Rates are highest in the 16-24-year-old age group. Untreated it can be a significant cause of morbidity. At least 50% of men and 70% of women with C. trachomatis are asymptomatic. STUDY AIMS: The aims of the study were: To determine the prevalence of C. trachomatis. To determine the success of our referral policy to genitourinary medicine (GU clinic). To determine the characteristics of the population with C. trachomatis. To estimate the level of recognition of 'chlamydia' as a concept. PARTICIPANTS: Attendees at our youth clinic between October 2001 and March 2002. METHODS: Ethical approval was obtained for this ongoing study. All attendees who were sexually active were asked to participate. An information leaflet was provided. Those who agreed to participate answered a questionnaire, which included a number of lifestyle questions, and provided a urine sample for C. trachomatis testing using a strand displacement assay. Positive results were forwarded to the GU clinic, which provided antibiotic therapy, contact tracing and follow-up. RESULT: The ongoing study has yielded 616 results with 73 positive (11.9%). To date 66 individuals (90%) have attended the GU clinic and 41 (50%) of the possible 82 partners have responded to contact tracing. CONCLUSION: Initial results show a high prevalence of C. trachomatis. There is a low condom usage despite a reasonable level of awareness of 'chlamydia'. Contact tracing has not been as successful as anticipated. When the study is complete, various service provision questions will need to be answered, such as the ability to treat the disease in a dedicated youth clinic, making urine testing for C. trachomatis by strand displacement assay (SDA) more widespread, the problem of follow-up, screening for associated sexually transmitted infections (STIs) and contact tracing in a relatively less mobile and less empowered population.
Fluorescence Polarization Assays (FPAs) have been shown to have great utility in the detection of infectious diseases. Examples are presented of the use of O-polysaccharides (OPSs) for the detection of antibodies in serum, whole milk and whole blood to gram negative organisms (Brucella spp., Salmonella spp.). The use of proteins and peptides are also described for the detection of Mycobacterium bovis and Equine Infectious Anemia Virus. Fluorescence Polarization Inhibition Assays (FPIAs) are discussed for the specific and sensitive detection and quantitation of Salmonella spp. cells from culture. An example of the detection of enterohemorrhagic E. coli (EHECS) by Strand Displacement Amplification (SDA), coupled with FP, down to the single cell level, within thirty minutes, is described.
OBJECTIVE: To evaluate the activity of Kupffer cells (KC) of control neonatal pigs and neonatal pigs treated with endotoxin and to compare activity of KC with that of pulmonary alveolar macrophages (PAM). SAMPLE POPULATION: Kupffer cells and PAM obtained from 24 neonatal pigs (7 to 10 days old). PROCEDURE: Pairs (n = 7) of littermates served as treated (lipopolysaccharide [LPS]) or untreated pigs. Pigs were euthanatized 24 hours after treatment, and cells were isolated. Cells were obtained from 10 other neonatal pigs for other assays. Functional activity of cells was evaluated by use of in vitro assays to evaluate bactericidal activity, phagocytosis, and production of superoxide anion (SOA), nitric oxide (NO), and tumor necrosis factor-alpha (TNF-alpha). Each assay was repeated on cells obtained from 4 to 6 pigs. RESULTS: Phagocytic activity was similar in KC and PAM, but bactericidal activity and production of SDA and TNF-alpha was lower in KC. Neither KC nor PAM produced NO in response to LPS stimulation. Phagocytosis, bactericidal activity, and production of SOA were enhanced for KC obtained from neonatal pigs treated with LPS. The PAM from LPS-treated neonatal pigs had similar bactericidal activity to PAM obtained from untreated pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Functional capacity of KC is affected by endotoxin. This provides additional information of the role the liver plays in immune surveillance. In addition, the response of KC in neonatal pigs exposed to endotoxin is of value for understanding gram-negative bacterial sepsis, which is a major cause of mortality in neonatal pigs.
Cyclosporine (CS) is a potent immunosuppressive agent which under some circumstances paradoxically augments DTH responses, aggravates some autoimmune diseases, and induces specific forms of autoimmunity. The enhancement of DTH and other immune responses is closely related to the timing of CS administration relative to immunization. CS inhibits IL-2 production (and several other lymphokines) at a pretranscriptional level, but does not usually prevent the antigen-specific priming of T cells, such that T cells may be poised to respond as soon as CS is withdrawn. Thus, accelerated GVHD and allograft rejection may occur after withdrawal of CS. CS has been shown to aggravate and/or induce relapse in several autoimmune diseases including collagen-induced arthritis, EAE, autoimmune thyroiditis, uveitis in SDA chickens, and an autoimmune form of myocarditis in mice. CS may enhance immune responses by inactivating suppressor cells, by altering Th1/Th2 antagonism (e.g., CS promotes a protective Th1-type response in BALB/c mice infected with Leishmania major), or by promoting T cell activation through a CS-resistant IL-2-independent T cell activation/differentiation pathway. At least three forms of CS-induced autoimmunity have been described: Syngeneic or autologous GVHD which occurs in CS-treated syngeneic or autologous bone marrow transplant recipients after CS is withdrawn in rats, mice, and humans; a systemic autoimmune disease with polyarthritis and glomerulonephritis which occurs in irradiated CBA/N mice treated with CS; and organ-specific autoimmune diseases which occur in mice treated with CS during the neonatal period. The precise mechanisms by which CS induces these autoimmune diseases are not clear, however, CS affects immune tolerance at three levels. CS induces thymic medullary involution with loss of medullary Ia+ cells, and appears to at least partially block the transition from double positive (CD4+CD8+) to single positive (mature type) thymocytes. In syngeneic bone marrow chimeras, CS appears to inhibit the intrathymic deletion of clones with relatively low affinity, but not those with high affinity, to self antigens. CS appears to inhibit the action of suppressor T cells which normally maintain an innate form of resistance to autoimmunity. Finally, CS has been shown to prevent the development of T cell clonal anergy. There is redundancy in immune tolerance mechanisms, i.e., clonal deletion, clonal anergy, and suppressor cells can each maintain tolerance to similar antigens, such that it is likely that CS must cripple more than one tolerance mechanism for autoimmunity to occur.
OBJECTIVE: To determine the various oral Candidal species among healthy Malaysian adults. DESIGN: Case-control study. PLACE AND DURATION OF STUDY: This study was collaborated between the Department of Medical Microbiology, Faculty of Medicine and Department of Oral Pathology, Oral Medicine and Periodontology, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia, between September 2002 till January 2004. PATIENTS AND METHODS: One hundred adults (50 smokers and 50 non-smokers), aged between 40 and 70 years were studied. Swabs and carbohydrate assimilation (Saboraud Dextrose Agar, Corn Meal Agar, API 20C AUX System) were performed. Specimens were collected from dorsum of the tongue, buccal mucosa and commissures (right and left each). Colony forms were established by positive colony forming units, on SDA medium (24-48 hours). Germ tube test for (true/pseudohyphae) growth was done on Corn Meal Agar Medium. Candida biotypes were evaluated by API 20C AUX system, which had a numerical 7 digit profile, added to evaluate a definite Candida species. RESULTS: Thirty-five percent of Malaysian adults harbored Candida intraorally. Candidal species identified among 100 subjects had C. albicans (27) 77%, C. glabrata (3) 8%, C. famata, C. tropicalis, C. krusei, C. lusitaniae and C. guilliermondii (1) 3% each. Thirty-three positive cases comprised of 35 species i.e. two cases had two species each. Fifty seven percent of these were smokers and 43% non-smokers. These included 40% Chinese, 36% Malays and 24% Indians. Species were, however, not specified according to intra-oral sites i.e. buccal, commissural mucosa and dorsum of tongue. CONCLUSION: On this series C. albicans is the most common species found in the oral cavity of Malaysian adults. It is equally frequent in smokers and non-smokers, but showed a predilection for the ethnic Chinese group.
Quantitative EEG was used to reveal the specific features of its amplitude-frequency parameters and topography in patients with mild dementia of different genesis versus healthy elderly individuals. All four study groups of patients differ in EEC relative spectral density, they also differ from the healthy persons while alpharhythm was suppressed in Alzheimer's disease (AD) and vascular dementia (VD) and there was slower alpha-rhythm and higher theta-activity in senile AD (SAD). Patients with VD were characterized by desynchronized EEG. Factorial and stepwise discriminant analyses of EEG parameters showed that the control group greatly differed from patients with mild dementias. Patient groups mainly with atrophic atrophic AD and SAD or vascular (VD and mixed) dementia were also different whereas the mixed vascular-atrophic group is intermediate in EEC parameters between the VD group and the partially overlapped AD and SDA patient groups. Quantitative EEC data may be thus used for differential diagnosis to optimize therapy and prognosis even in mild dementia.
BACKGROUND AND PURPOSE: Rat coronaviruses (RCV) are highly infectious and spread rapidly through laboratory rat colonies, causing sneezing, nasal and ocular discharges, photophobia, and cervical swelling. Current diagnostic methods include serologic testing and histologic examination. During a recent rat coronavirus outbreak, we tested a rapid, noninvasive method of RCV diagnosis that involved use of reverse transcriptase-polymerase chain reaction (RT-PCR) analysis to detect RCV RNA on cages housing infected rats. METHODS: The RT-PCR was used to detect RCV RNA in tissues from infected rats and on cages housing infected rats and to amplify portions of the RCV N, M, and S genes for molecular characterization. RESULTS: The RT-PCR detected RCV RNA on cages and in tissues from infected rats. The RCV-NJ N gene is most closely related to the MHV-Y N gene. The M proteins of RCV-NJ and RCV-SDA are 99% homologous, and the six RCV S protein fragments are 97 to 100% homologous. CONCLUSIONS: Use of RT-PCR with cage-swab specimens was capable of diagnosing RCV infection in and viral excretion from rats. Additionally, molecular characterization of the N, M, and S genes of RCV-NJ provided baseline information that can be used in performing further epidemiologic studies.
OBJECTIVE: The aim of the study was to evaluate the effectiveness of a sugar-free medicines campaign using suitable prescribing and purchasing indicators. BASIC RESEARCH DESIGN: Quantitative analysis of sugar-free medicine use, before (1995) and after (1996) the campaign. CLINICAL SETTING: Two test and two control districts in north east England. PARTICIPANTS: General medical practitioners (GPs) and community pharmacists. INTERVENTION: A 12-month campaign involving development and distribution of information packs designed to increase the proportion of prescriptions dispensed sugar-free for paediatric use. MAIN OUTCOME MEASURES: Changes in the proportion of the following which were sugar-free: (a) for target prescribed medicines (i) number of prescriptions dispensed. (ii) number of prescribed daily amounts (PDA); (b) for target over the counter (OTC) medicines, (i) number of bottles sold, (ii) number of standard daily amounts (SDA) sold. RESULTS: Quantitative analysis of prescriptions and OTC sales showed statistically highly significant changes towards sugar-free prescribing and dispensing of prescribed medicines but only small increases in the proportion of some sugar-free OTC sales. CONCLUSIONS: The impact of the campaign was shown to be greatest in changing prescribing habits of GPs with less effect on OTC medicine use. The outcome measures used were suitable for quantitative evaluation of the campaign. POST-CAMPAIGN DEVELOPMENT: Sustainable changes in GPs' prescribing behaviour can be facilitated by software suppliers' modifications to computing software used for prescription writing.
The various parameters proposed in Norm 0.20/95 of Catalunya (Spain) for the microbiological analysis of natural corks for sparkling wines were evaluated. The best results were obtained through the use of 1/4 Ringer's solution or saline for rinsing with an agitation time of 30 min, and an agitation speed of 150-200 rpm. Tryptone soya agar (TSA) and Sabouraud dextrose agar (SDA) were used as a culture medium for the bacteria and fungi, respectively, and a cultivation time of 48 h and incubation temperatures of 37 +/- 2 degrees C for bacteria and 28 degrees C for yeast and filamentous fungi.
We prospectively evaluated the usefulness of adenosine deaminase [ADA] estimation in the diagnosis of tuberculosis [TB] pleural effusion. Seventy five subjects with pleural effusion were studied. Forty eight of them had TB pleural effusion [M:F: 37:11; mean age 33 +/- 14.4 years range 17-76] and the remaining 27 had pleural effusion due to causes other than TB [non-TB group] [M:F: 19:8; mean age 47.3 +/- 16.5 years; range 17-75]. Pleural fluid [PF] ADA levels were significantly higher in TB (n=48; mean 95.8 +/- 57.5 IU/L) compared with non-TB group (n=27; mean 30.7 +/- 27.2 IU/L) [p<0.001]. Serum ADA [S-ADA] levels were also significantly higher in TB (n=45; mean 39.6 +/- 18.3 IU/L) compared with non-TB group (n=26; mean 18.0 +/- 13.7 IU/L) [p<0.001]. PF-ADA levels were higher compared to S-SDA in TB (p <0.001) and non-TB groups [p<0.01]. Using a cut off of 35 IU/L, the sensitivity and specificity of PF-ADA in the diagnosis of TB was computed to be 83.3% and 66.6% respectively. At a cut-off level of 100 IU/L, PF-ADA was found to have a sensitivity 40% and specificity 100%. From this study it is concluded that, using 100 IU/L as the cut-off, it is possible to avoid pleural biopsy to ascertain the diagnosis of TB in as much as 40% of the patients.
BACKGROUND: Acute hypoxia tolerance varies substantially among healthy individuals. We hypothesized that this variability results from a difference in autonomic (ANS) response to hypoxic stress. METHODS: Peripheral oxygen saturation, respiration and ECG were recorded from 21 healthy subjects (age, 29 +/- 7 yr) in an altitude chamber during normoxia, severe hypoxia (282 mm Hg), and mild hypoxia (360 mm Hg). Cardiovascular control was assessed by time-frequency decomposition of the heart rate signal applying the Selective Discrete Transform Algorithm (SDA). This procedure uses a variable time window, thus providing reliable physiological data even during transient states. Autonomic activity was quantified by power spectral density integrals over a 3-dimensional time-dependent spectral distribution of heart rate fluctuations. RESULTS: Subjects who had slower peripheral oxygen desaturation during severe hypoxia onset (mean 92.9 vs. 58.4 s) were those who displayed higher ANS activity in all ambient states, namely normoxia and hypoxia. These same subjects withstood hypoxia for significantly longer time periods (mean 313 vs. 244 s). CONCLUSION: Improved hypoxia tolerance is linked to enhanced autonomic activity, involving a better management of peripheral blood flow.
BACKGROUND: CHROMagar Candida (CAC) is a new chromogenic medium for the presumptive identification of clinically-important yeast isolates. A yeast biochemical card (YBC), a part of the Vitek system is an automatic method for the identification of clinically-important yeast isolates. We conducted a comparison of these two methods with a traditional biochemical method in order to choose a rapid and accurate technique for yeast identification. METHODS: All yeast isolates were inoculated onto Sabourand dextrose agar (SDA) and CAC, and incubated at 30 degrees C for 48 hours. All isolates were simultaneously tested using traditional biochemical methods and the yeast biochemical card from the Vitek system. RESULTS: We evaluated 235 yeast isolates from clinical specimens, including 89 Candida albicans, 47 Candida tropicalis, 43 Candida glabrata, six Trichosporon beigelii, and five Candida krusei in addition to 45 isolates of other yeast species. Isolates were presumptively identified on the basis of colony color and appearance on CAC medium. These observations were compared with a traditional biochemical yeast-identification method and also with YBC from the Vitek system. For five commonly-isolated species (Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei and Trichosporon beigelii), agreement among the CAC medium, YBC method and traditional biochemical method were 98.9% (187/189), 96.3% (182/189), 100% (189/189), respectively. CONCLUSIONS: From our comparison, the CAC medium is a convenient and economic method to identify five commonly-noted yeast species, and the YBC method warrants a greater cost and requires a longer period of time to obtain reliable results.
The purpose of this in vitro study was to investigate the effects of microbial colonization on different crown or fixed bridge-used restorative materials and discuss the standards for evaluating those materials microscopically. Four kinds of restorative materials such as SDA-II type medium alloy, ceramic-fused-to-metal material, heat cure composite resin for crown restoration, Plat castable ceramics and four species of precedent subgingival flora (A. viscosus, F. nucleterm, C. ochracea, S. sanguis) were used. The material specimen had been continuously anaerobic cultured with the experimental bacterial fluid for 3, 7 and 14 days respectively, then the OD values of the specimenwashing fluid which could stand for the biomass of colonized bacteria on the surfaces of materials were measured. The results indicated that the quantity of precedent subgingival flora colonization on the surfaces of different restorative materials varied among different species of bacteria, which might concern with the surface structure, composition, property of anticorrosion and antisoluability of those materials.
The aim of the study was to establish the importance of KOH mount in rapid diagnosis of fungal corneal ulcers. Corneal scrapings were collected from 50 patients with corneal ulcers where clinical features suggested fungal aetiology and ulcers showed no signs of healing on initial treatment. Corneal scrapings were subjected to wet mount with 10% KOH, Gram's stain, and culture on SDA media. Wet mount showed fungal elements in 34 cases (68%) and were classified according to morphology. Gram's stain of corneal scrapings did not provide much help. Fungal culture was positive in 9 (26.4%) out of 34 cases. Specific antifungal treatment was instituted in all cases that showed positive wet mount without waiting for culture reports and on follow-up all these 34 patients showed improvement. Thus wet mount with 10% KOH can be relied upon as the singlemost important screening tool for rapid diagnosis of fungal corneal ulcer and treatment should be dispensed on its basis.
OBJECTIVES: To improve the latency change estimation of evoked potentials (EP) under the lower order alpha-stable noise conditions by proposing and analyzing a new adaptive EP latency change detection algorithm (referred to as the NLST). METHODS: The NLST algorithm is based on the fractional lower order moment and the nonlinear transform for the error function. The computer simulation and data analysis verify the robustness of the new algorithm. RESULTS: The theoretical analysis shows that the iteration equation of the NLST transforms the lower order alpha-stable process en (k) into a second order moment process by a nonlinear transform. The simulations and the data analysis showed the robustness of the NLST under the lower order alpha-stable noise conditions. CONCLUSIONS: The new algorithm is robust under the lower order alpha-stable noise conditions, and it also provides a better performance than the DLMS, DLMP and SDA algorithms without the need to estimate the alpha value of the EP signals and noises.
Depending on sequence and backbone structure, DNA can inhibit as well as stimulate immune responses. As previously shown, single-base phosphorothioate (Ps) oligodeoxynucleotides (ODN) can inhibit murine macrophage activation. To determine whether these compounds can also affect dendritic cells (DC), the effects of 30-mer Ps ODN (SdA, SdT, SdG, and SdC) on DC activation were assessed in an in vitro system. With DC preparations obtained from murine bone marrow cultured in granulocyte macrophage-colony stimulating factor, the Ps ODN blocked the production of interleukin-12 and nitric oxide induced by bacterial DNA, an immunostimulatory cytosine phosphate guanosine dinucleotide (CpG) ODN and lipopolysaccharide (LPS). Furthermore, these compounds inhibited up-regulation of costimulatory molecules CD40 and CD86 as well as major histocompatibility complex-II molecules, indicating an effect on DC maturation. Although the Ps ODN limited uptake of CpG ODN as assessed by flow cytometry, the Ps ODN did not affect LPS uptake, suggesting that these compounds inhibit DC responses by effects on downstream signaling pathways. Together, these observations extend the range of action of inhibitory ODN to DC and suggest a role of these compounds as immunomodulatory agents.
OBJECTIVE: To investigate the epidemiologic features of disseminated histoplasmosis (PDH) in Hubei province. METHODS: Bone marrow smears of 12 patients diagnosed as Kala-azer in Hubei province including 4 patients in Jingsan, 2 patients in Shashi and each 1 in Yichang, Jinmen, Zhongxiang, Luotian, Xianning and Guanghua respectively were re-examed under microscope. Peripheral blood and bone marrow smears of several patients were detected. After inoculated the bone marrow, peripheral blood, liver and spleen tissue of patients in MLI, the single colony was trans-inoculated in BHIB, SDA and CMA and incubated at 25 degrees C and 35 degrees C. Bone marrow, peripheral blood and bacterial fluid of yeast-phase Histoplasma capsulatum (H.cap) were injected into the abdominal cavity of Kunming mice and nude mice. When symptoms and signs developed, the spleen tissue was separated, then observed under microscope and cultured. Mycelium-phase and Yeast-phase H.cap were inoculated in urase and gelatin medium, then incubated at 25 degrees C and 35 degrees C. Histoplasmin was injected subcutaneously into patients, and then followed for 48 - 72 hours. Amphotericin B was selected to treat the PDH patients. RESULTS: Moriform cell cluster and sausage-shaped cell were not observed in mononuclear-macrophages in the bone marrow smears from 12 patients. Leishman-Donovan body was found only in one patient. There wasn't kinetoplast in the cellular plasm of spores in 11 patients and no transeptae was found. The reaction of H.cap to urease was positive and H.cap did not liquefy the gelatin. It appeared to be mycelium-phase at 25 degrees C but no penicillus and catenulate conidia was found. The characteristic denticle macroconidia was observed but produced red coloring matter. It also appeared to be yeast-phase at 35 degrees C. Yeast-phase spores were observed under microscope. No sausage-shaped spore and transeptae were identified. H.cap could be acquired in the spleen tissue in Kunming mice and nude mice. Bacterium forms, characteristics under microscope and biochemical reaction of mycelium-phase and yeast-phase H.cap were different from some other kinds of dimorphic fungi such as Penicillium marneffei and Histoplasm duboisii etc. CONCLUSION: There were scattered epidemics of PDH in Hubei province. The detection rate of PDH was higher in the southeast area then in the northwest area. The golden standards of clinic diagnosis were mycological culture and inoculation to animals. Amphotericin B was necommended as the first choice for treatment.