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PubMed1999

Comparison of the pathogenicity in rats of rat coronaviruses of different neutralization groups.

BACKGROUND AND PURPOSE: Rat coronaviruses (RCVs) are common natural pathogens of rats that cause clinical illness, necrosis, and inflammation of respiratory, salivary, and lacrimal organs. The aim of the study was to determine whether antigenically different strains of RCV vary in their pathogenic potential in rats. METHODS: Neutralization groups were identified by use of RCV strain-specific antisera. Sprague Dawley rats were inoculated oronasally with RCV-SDA, RCV-BCMM, or RCV-W. Histologic examination, immunohistochemical analysis, and reverse transcriptase-polymerase chain reaction analysis were performed on tissues from infected rats. RESULTS: Clinical illness was not evident in any of the inoculated rats. The RCV-SDA strain caused mild lesions in the exorbital gland of one rat. The RCV-BCMM strain caused severe lesions in the Harderian and parotid glands and mild lesions in the exorbital glands, lungs, and nasal mucosa. The RCV-W strain caused severe lesions in the Harderian, exorbital, and parotid glands and mild lesions in the submandibular glands, lungs, and nasal mucosa. The RNA concentration was highest in the Harderian, parotid, and exorbital glands of RCV-BCMM- and RCV-W-infected rats at postinoculation day 7. CONCLUSIONS: Although RCV-SDA, RCV-BCMM, and RCV-W caused different degrees and patterns of lesions, neutralization groups are not useful for predicting the pathogenic potential of a new RCV isolate.

Animals↗
PubMed2000

Pathogen analysis and genetic predisposition testing using microelectronic arrays and isothermal amplification.

BACKGROUND: A simple yet powerful tool for providing for rapid gene identification in the clinic would be the combination of isothermal gene amplification with electronic microchip analysis. This is a first report of such a union of these technologies. METHODS: The first assay demonstrates discrimination between four bacterial pathogens. For this, one portion of the bacterial 16S rRNA gene encompassing a microheterogeneous region was isothermally amplified using Strand Displacement Amplification (SDA). Type identification was then made by "sandwich" assay format either using selective electronic hybridization of amplicons to sequence-specific capture oligonucleotides and a universal, fluorescently labeled reporter oligonucleotide, or, alternatively, sequence-specific reporters and a universal capture oligonucleotide. The second assay tested for the presence or absence of the Factor V Leiden point mutation using DNA obtained from 18 patients in a blind assay. For this, allele-specific SDA was developed. Following amplification using a sense-biotinylated primer and either the corresponding antisense wild type or mutant primer, multiple patient amplicons were targeted to specified locations on the microarray and visualized using a fluorescently labeled reporter oligonucleotide. Positive signals were scored as greater than or equal to two times the background. RESULTS: Bacterial type-specific signals were between 3- to 10-fold greater than nonspecific in both assay formats. Using allele-specific SDA, 100% agreement was observed between PAGE analysis, microarray results, and clinical diagnosis in Factor V mutation analysis. CONCLUSIONS: We demonstrated two model clinical assays combining amplified materials and microelectronic arrays, one potentially suitable for pathogen screening and the other for a deleterious genetic mutation.

Bacteria↗
PubMed2002

[Effects of cryotreat on the corrosion resistance of the medium melting-point castable alloy].

OBJECTIVE: The purpose of this study was to investigate effects of cryotreat on the corrosion resistance of SDA-II medium melting-point castable alloy. METHODS: 12 samples were divided into three groups, including the control group, the cryotreated group and the cryotreated & post-cryogenic treated group. The cryotreating process was firstly dropping the temperature from room temperature to -170 degrees C with dropping rate 3 degrees C/min, maintaining -170 degrees C for 2 hrs, and then returning to ambient temperature with an increasing rate of 1 degree C/min. The post-cryogenic process was maintaining the samples in 300 degrees C for 1 hr. Finally, all samples were subjected to the electrochemical potentiodynamic polarization after immersed in the artificial saliva (ISO/TR 10271: 1993 (E), pH 6.8) for 24 hrs at ambient temperature. The exposing area of each sample was 10 mm x 10 mm. The electrochemical tests were carried out in the artificial saliva under 37 degrees C, with the scanning rate of 20 mV/min, starting potential -200 mV below OCP and stopping current 10(-3) A. The corrosion currents (Icorr), the pitting corrosion potentials (Eb) and the potentiodynamic polarization curves were observed using the 283 electrochemical corrosion measuring system automatically. RESULTS: For SDA-II medium melting-point castable alloy, cryotreat reduced corrosion rate of the alloy, but had no effect on the pitting corrosion potential. As far as the post-cryogenic treatment, it didn't affect the corrosion resistance. CONCLUSION: The results suggested that cryotreat was an effective procedure in enhancing the corrosion resistance of SDA-II medium melting-point castable alloy, and it may be applicable in prosthetic dentistry.

Cold Temperature↗
PubMed2003

[The effect of cryotreat on the wear resistance of the medium melting-point castable alloy].

OBJECTIVE: The purpose of this study was to introduce cryotreat technique into prosthetic dentistry by investigating the wear resistance changes of SDA-II medium melting-point castable alloy before and after cryotreat. METHODS: 15 samples were divided into three groups, i.e. control group, cryotreat group, cryotreated plus post-cryogenic treat group. Weight loss measurement was used to examine the wear resistance. RESULTS: For SDA-II medium melting-point castable alloy, cryotreat effectively improved its wearing resistance; so far as the wear resistance was concerned, the post-cryogenic treatment was unnecessary. CONCLUSION: The results obtained above suggest that cryotreat was an effective method in enhancing the wear resistance of SDA-II medium melting-point castable alloy.

Cold Temperature↗
PubMed1996

A new form of polyagglutination related to Cad.

Four phenotypes of Cad (Cad 1-4) have been characterized by a continuum of polyagglutinability and reactivity with lectins, with the strongest Cad+ red blood cells (RBCs) being polyagglutinable because of the presence of anti-Cad (anti-Sda) in most normal sera. Over a period of 7 years, a French male blood donor's RBCs demonstrated polyagglutinability with 50 percent to 70 percent of normal adult sera. The reactivity was characteristic of anti-Sda (refractile agglutination at 4 degrees C, 20 degrees C, 37 degrees C, and anti-human globulin test), and was inhibitable by two examples of Sd(a+) urine, but not by Sd(a-) urine or dialysate from Sd(a+) urine. The donor's RBCs reacted 1+ with Glycine max, but did not react with Dolichos biflorus, Leonurus cardiaca, Salvia horminum, or Arachis hypogaea. The first four of these lectins were reactive with five of five Cad+ RBCs, including one example of Cad 4 RBCs. Polybrene(R) aggregated the donor RBCs. Dilutions of nine samples of anti-Sda reacted more strongly with the donor RBCs than with normal RBCs. Even though lectin studies failed to classify this donor's RBCs as Cad, the persistent polyagglutinability and serologic characteristics are consistent with Cad and demonstrate the heterogeneity of this antigen.

Journal Article↗
PubMed2005

Performance of strand displacement amplification assay in the detection of Chlamydia trachomatis and Neisseria gonorrhoeae.

This study is a critical analysis of certain amplification assays for detecting Chlamydia trachomatis and Neisseria gonorrhoeae infections which have demonstrated that the plasmid-free variant of C. trachomatis is frequently responsible for infection in our patients. Specifically, we evaluated the performance of the strand displacement amplification (SDA) assay in detecting either C. trachomatis or N. gonorrhoeae in 1,190 clinical samples, both urogenital and ocular, from 1,005 consecutive patients. The results obtained with the BDProbeTec ET System were compared with three referenced amplification methods for C. trachomatis (detecting the 16S rRNA gene, the omp1 gene and the plasmid of C. trachomatis) and with both the culture method as well as an amplification assay followed by genetic identification performed using the MicroSeq 500 16S ribosomal DNA-based system for N. gonorrhoeae. The sensitivity of SDA (76%) in detecting C. trachomatis is significantly low when compared with that of other molecular techniques employing 16S rDNA or omp1 as a target. The specificity of the methods for detecting C. trachomatis was excellent, ranging from 99.4 to 100%. Furthermore, the results of SDA in detecting N. gonorrhoeae also provided excellent results (100% specificity and sensitivity).

Adult↗
PubMed1991

[Psychopathological mechanisms of socially dangerous acts and the problem of responsibility].

The authors discuss the problems of forensic-psychiatric expert examination related to the responsibility of persons suffering from different mental disorders. In addition to the assessment of the clinical picture of a mental disease, it is necessary to take into consideration (as fully as possible) the genesis and character of the committed socially dangerous action (SDA). For this purpose it is recommended that the concept of the psychopathological mechanisms of SDA worded by the authors and their classification be made use of. The establishment in a mental patient of one of the distinguished mechanisms of SDA allows the differentiation of the intellectual and volitional signs of the judicial (psychological) criterion and scientific basing of the conclusion as to the patient's irresponsibility.

Crime↗
PubMed1985

Epizootiological observations of natural and experimental infection with sialodacryoadenitis virus in rats.

The epizootiology of sialodacryoadenitis (SDA) was studied in experimentally and naturally infected rats. The infectivity of SDA virus (SDAV) in intranasally infected rats was lost by seven days after infection as determined by contact transmission. After experimental infection, SN antibody appeared earlier and titers were detectable longer than CF antibody. The prevalence of SN antibody-positive rats in naturally infected colonies remained high, whereas an increase in the prevalence of CF antibody-positive rats appeared to coincide with the introduction or resurgence of SDAV. A SDAV-free colony was established by allowing recovered dams to litter in a separate room. A spontaneous cessation of SDAV infection also was observed in an enzootically-infected colony. Clinical observations indicated that SDA can occur as a mild or asymptomatic disease, and that its clinical expression may vary from one inbred strain to another.

Animals↗
PubMed1985

Mortality in Japanese with life-styles similar to Seventh-Day Adventists: strategy for risk reduction by life-style modification.

Using 16 years of follow-up results of a prospective cohort study for 122,261 men, 95% of the census population, aged 40 years and older in 29 Health Center Districts in Japan as subjects, we compared the age-standardized mortality rates for cancer of each site and other causes of death in Japanese with life-styles similar to those of Seventh-Day Adventists (SDA), i.e., no smoking, no drinking, no meat consumption daily and eating green and yellow vegetables daily, with those of Japanese with opposite life-styles. Compared with the latter Japanese, the risks were one-fifth or less in Japanese with SDA-like life-styles for cancers of the mouth, pharynx, esophagus, and lung, and subarachnoid hemorrhage. Risks were less than one-half for cancers of all sites, stomach, and liver, and for peptic ulcer and heart disease. As a single factor, the addition of daily smoking was observed to elevate the risk most strikingly in Japanese who followed SDA life patterns. Influences of further addition of habits of daily drinking of alcohol and dietary changes were significant for cancers of the esophagus, liver, and bladder and other selected diseases. Strategies for cancer prevention by means of life-style modification, e.g., increased consumption of green and yellow vegetables, were discussed.

Adult↗
PubMed1985

Cholesterol side chain cleavage in rat adrenal supported by outer mitochondrial membrane NADH-semidehydroascorbate reductase.

Rat adrenal mitochondria have an active rotenone-insensitive outer mitochondrial membrane NADH-semidehydroascorbate (NADH-SDA) reductase which supports cholesterol side chain cleavage at a rate equal to that supported by malate. Side chain cleavage activity supported by both of these electron donor systems is equally inhibited by cycloheximide. Catalase or butylated hydroxyanisole are required for the NADH-SDA reductase-supported cholesterol side chain cleavage. This requirement can be removed by briefly subjecting the mitochondrial preparations to -20 degrees C. Ascorbic acid alone or with malate is either inhibitory or has no effect on side chain cleavage activity. These observations demonstrate that outer mitochondrial membrane NADH-SDA reductase in rat adrenal functions to provide cytoplasmic reducing equivalents to intramitochondrial cytochrome P-450scc and provides a new explanation for the function of ascorbic acid in corticosteroidogenesis.

Adrenal Glands↗
PubMed1994

[Investigation of treating steroids dependent asthmatic patients with kidney-tonifying herbs and high dose beclomethasone dipropionate aerosol].

The steroids dependent asthmatic(SDA) patients need prolonged oral steroids administration. Because of the significant adverse effects with long-term use of steroids, especially the damage of the hypothalamus-pituitary-adrenocortical(HPA) axis, long-term steroids administration should be avoided or withdrawn gradually. Many modes of withdrawing the oral steroids have be used. But, the results were not satisfactory. So far, recently, investigations have revealed that high dose corticosteroids inhalation, such as beclomethasone dipropionate (BDP), could provide an effective alternative; they were delivered locally in the airway and had minimal systemic absorption as well as side effects. On the other hand, it has been shown that the Kidney-Tonifying herbs (KTH) exert protective effect on the adrenocortical cells of the SDA patients by suppressing the exogenous steroids and could regulate the disorders in different levels on HPA axis in the patients. In order to provide the more effective alternative therapy for the patients, 30 SDA patients replaced their oral steroids with KTH and high dose BDP (1500 micrograms/day) inhalation were investigated. The changes of clinical features, pulmonary function and the Synacthen Test were followed up. The results showed that about 70% of the patients replaced their oral steroids successfully with the new therapy, the clinical features were improved in 16% of the patients and the ineffective rate was only 6.6%; the results also revealed that, after replacement, the pulmonary function of the patients were improved (P < 0.05); the data of Synacthen Test indicated, after alternative therapy for 3-6 months, the damaged reserve power and secretive ability of adrenal cortex of the patients were also improved partially (P < 0.001).

Aerosols↗
PubMed1996

Detection of Mycobacterium tuberculosis DNA with thermophilic strand displacement amplification and fluorescence polarization.

Strand displacement amplification (SDA) is an isothermal, in vitro method for diagnostics that amplifies a target DNA sequence by using a restriction enzyme and DNA polymerase. We have combined a new thermophilic form of SDA that involves restriction enzyme BsoBI and polymerase exo-Bca with fluorescence polarization for detection of Mycobacterium tuberculosis DNA by using the IS6110 insertion element as the target sequence. A 5'-fluorescein-labeled oligodeoxynucleotide detector probe hybridizes to the amplified product as it rises in concentration during SDA, and the single- to double-stranded conversion is monitored through an increase in fluorescence polarization. The associated change in polarization upon amplification of the target sequence is enhanced by specific polymerase binding to the double-stranded detector probe. Fewer than 10 M. tuberculosis genomes can be amplified and detected with an extremely simple protocol that takes only 20 min and uses relatively simple instrumentation and reagents, all of which can be purchased off-the-shelf.

DNA Primers↗
PubMed1995

Meta-[123I]iodobenzylguanidine single photon emission computed tomography in chemodectomas.

Chemodectoma is a rare, slow growing neoplasm with local aggressiveness and a high rate of residuals after surgery, arising from paraganglionic tissue (neural crest) and therefore able to take up Metaiodobenzylguanidine (MIBG). The aim of this study was to evaluate the diagnostic accuracy of 123I-MIBG Single Photon Emission Computed Tomography (SPECT) in comparison to Selective Digital Angiography (SDA), Computed Tomography (CT) and Magnetic Resonance Imaging (MRI). We studied 50 patients (41 women, 9 men) mean age 56 years (range 14-79), with diagnostic suspicion of chemodectoma (CH). There were 2 groups of patients: Group A (26 patients) examined before Surgery and Group B (24 patients) examined after surgery. SPECT of the head and neck region was performed 4 hours after i.v. administration of 185 MBq of 123I-MIBG. All patients were pre-treated with iodine solution. We performed a qualitative evaluation of the reconstructed slices. In Group A, all patients with CH (24) showed accumulation of MIBG in agreement with SDA, CT and MRI: 2 patients (histologically one had a cordoma, the other metastasis of papillary thyroid carcinoma), did not show any accumulation of 123I-MIBG. In Group B, 6 patients showed accumulation of 123I-MIBG in local residuals, and 9 were negative. Six patients with a lesion smaller than 1.5 cm were negative. Three patients had a positive scan but no lesion on SDA, CT and MRI. 123I-MIBG SPECT proved to be useful procedure in the diagnosis of untreated CH. During follow-up after surgery this procedure may assume a role in the perspective of radiometabolic therapy with 131I-MIBG.

3-Iodobenzylguanidine↗
PubMed1998

Saprophytic and cycloheximide resistant fungi isolated from golden hamster.

Healthy hair samples from golden hamsters were examined for the presence of dermatophytes and non-dermatophytes using baiting technique and direct inoculation. Thirty-four species and 2 varieties attributed to 17 genera were recovered. Paecilomyces variotii (isolated from 84.4% of the examined hair) and Aspergillus niger (81.3%) were the more frequent isolates on Sabouraud's dextrose agar (SDA) without cycloheximide. Our results have clearly demonstrated that the hair of hamster was free from true dermatophytes. Using the dilution plate method many fungal species were isolated from cage material (7 genera and 10 species + 1 variety); from faeces (10 genera and 17 species); from standard chow (3 genera and 6 species) of hamster. P. variotii which was the most frequent fungus in the preceding 3 substrates was completely absent in the presence of cycloheximide in SDA. The present study has demonstrated for the first time the isolation of Trichophyton rubrum from hamster faeces. Also, several saprophytic and cycloheximide resistant fungi were isolated. In the air of hamster cage Cladosporium cladosporioides, Penicillium chrysogenum, Alternaria alternata and Scopulariopsis brevicaulis were the most dominant species on SDA with or without cycloheximide. Using the agar diffusion method, Aloe sap, onion oil, garlic bulb extract and aqueous leaf extracts of Andropogon citratus, Euphorbia sp. and Ruta graveolens were tested for their antifungal activity on 10 fungal species. It was observed that onion oil exhibited a high inhibitory effect against most of the tested fungi.

Animal Feed↗
PubMed2002

Projections of the posterodorsal preoptic nucleus and the lateral part of the posterodorsal medial amygdala in male gerbils, with emphasis on cells activated with ejaculation.

The posterodorsal preoptic nucleus (PdPN) and the lateral part of the posterodorsal medial amygdala (MeApd) express Fos with ejaculation in male gerbils. Ejaculation-activated cells participate in the PdPN and MeApd projections to each other and to the sexually dimorphic preoptic area (SDA), but those projections involve less than 20% of the activated PdPN cells and less than 50% of the activated MeApd cells. To identify other potential targets of ejaculation-activated cells, we traced PdPN and lateral MeApd outputs using biotinylated dextran amine. The principal part of the bed nucleus of the stria terminalis (BSTpr) and the anteroventral periventricular nucleus (AVPv) were labeled from both sites and were injected with Fluoro-Gold to determine whether PdPN and lateral MeApd cells that express Fos with ejaculation would be retrogradely labeled. Fluoro-Gold was also applied to the dorsomedial hypothalamus (DMH) and retrorubral field (RRF) because such injections label PdPN cells in rats. The PdPN-DMH projection is minimal in gerbils, involving few, if any, ejaculation-related cells. Ejaculation-activated PdPN cells project to the AVPv (43%), dorsal BSTpr (30%), and RRF (12%). Those in the lateral MeApd project to the dorsal BSTpr (43%) and AVPv (18%). When these percentages are combined with those for ejaculation-activated cells involved in the PdPN and lateral MeApd projections to each other and to the medial SDA, the totals reach 100%. Thus, every PdPN and MeApd cell activated with ejaculation may participate in one of these projections. Similar projections may contribute to the similar behavioral effects of the PdPN and MeApd.

Afferent Pathways↗
PubMed2002

Symbolic discriminant analysis of microarray data in autoimmune disease.

New laboratory technologies such as DNA microarrays have made it possible to measure the expression levels of thousands of genes simultaneously in a particular cell or tissue. The challenge for genetic epidemiologists will be to develop statistical and computational methods that are able to identify subsets of gene expression variables that classify and predict clinical endpoints. Linear discriminant analysis is a popular multivariate statistical approach for classification of observations into groups. This is because the theory is well described and the method is easy to implement and interpret. However, an important limitation is that linear discriminant functions need to be prespecified. To address this limitation and the limitation of linearity, we have developed symbolic discriminant analysis (SDA) for the automatic selection of gene expression variables and discriminant functions that can take any form. In the present study, we demonstrate that SDA is capable of identifying combinations of gene expression variables that are able to classify and predict autoimmune diseases.

Autoimmune Diseases↗
PubMed1994

Inhibition of the induction of nitric oxide synthase by spermine is modulated by aldehyde dehydrogenase.

The observation that spermine inhibits the endotoxin (lipopolysaccharide; LPS) induced production of nitric oxide (NO) in macrophages has been ascribed to the conversion of SP to active metabolites by the action of enzymes, such as diamine oxidases, found, for example, in bovine sera. Inhibitory effect is also observed with the oxidised metabolite of spermine, spermine dialdehyde (SDA). Inhibition appears to be at the level of induction of the inducible isoform of NO synthase (iNOS). Here we show that the activity of endogenous aldehyde dehydrogenase present in the cells influences the degree of inhibition seen with either spermine or SDA. Most significantly, inhibition of aldehyde dehydrogenase activity greatly increases (100 fold) the ability of spermine to inhibit the production of nitrite by LPS- induced macrophages. This is presumably by preserving aldehyde metabolites of spermine and thus increasing its action on the induction of iNOS. Thus, inhibition of aldehyde dehydrogenase activity in vitro or in vivo may be a useful approach to enhance the inhibitory effect of polyamines or polyamine aldehydes on iNOS induction.

Aldehyde Dehydrogenase↗
PubMed1995

Modelling viral evolution in vitro using exo- Klenow polymerase: continuous selection of strand displacement amplified DNA that binds an oligodeoxynucleotide to form a triple-helix.

Evolution comprises cycles of amplification, mutagenesis and selection. To study evolutionary phenomena, isothermal strand displacement amplification (SDA) of double-stranded DNA as an in vitro model for rolling-circle replication of viruses has been coupled to a positive selection procedure. First, two subsequent amplification reactions utilizing exo- Klenow polymerase were performed under direct observation using the fluorescent dye thiazole orange. Under the chosen conditions, the mutation rate was 1.5 x 10(-3) and 0.4 x 10(-3) for base substitutions and deletions, respectively. Then, a 16mer oligodeoxynucleotide with an acridine moiety coupled to its 5' end was used to select for double strands that retained their ability to form a triple-helix with the oligodeoxynucleotide. Conditions for triple-helix formation were chosen such that only 10 to 40% of the SDA products were allowed to bind the third strand. Non-denaturing polyacrylamide gel electrophoresis was used to separate triple-helices from unmodified double strands, and only triplex strands were used to initiate a new round of error-prone amplification and selection. Nine such rounds with about 270 molecular generations were performed. The final mutant spectrum was characterized and compared with those of amplification reactions without additional selection pressure. While without selection pressure base substitutions and deletions throughout the initial wild-type rapidly produce a diverse mutant distribution, the consensus after nine selection rounds clearly shows two mutational hotspot positions. Using gel shift assays and a newly developed non-radioactive DNase I footprinting technique, it could be shown that both the initial wild-type and the final consensus do not differ significantly in their triplex formation ability. As opposed to this, they do show different amplification efficiencies. The final consensus sequence is amplified with the highest rate in the exponential reaction phase, while the most abundant clone, which is characterized by two additional point deletions, is the sequence with the highest amplification rate in the linear growth phase.

Base Sequence↗