Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In this study, Sprague-Dawley rats were exposed in a TEM chamber to 20-MHz (HF-band) continuous-wave radiofrequency radiation (RFR) for 6 hr/day, 5 days/week up to 6 weeks. The average E-field intensity was 2686 +/- 164 V/m (mean +/- SD) and the calculated specific absorption rate was 0.3 W/kg. Randomly sampled rats killed on Days 8, 22, 39, and 42 after initiation of exposure showed no statistically significant differences from controls for body mass, spleen cell density, erythrocyte and leukocyte counts, hematocrit, hemoglobin, methemoglobin, erythrocyte fragility, bilirubin, creatinine, SGPT, alkaline phosphatase, calcium, sodium, potassium, and spleen cell chemiluminescence. Splenic mass differences were statistically significant (p less than 0.05) only on Day 22. Spleen to body mass ratios differed significantly between exposed and control groups on Days 22 and 39 (P less than 0.05 and P less than 0.025, respectively). Histologic examination of the rats revealed the successive accumulation of phagocytic cells, lymphoid proliferation, development of lesions, and tissue necrosis characteristic of respiratory mycoplasmosis. In a followup experiment, a separate set of rats was exposed for 6 weeks to identical levels of RFR. No significant differences were found in splenic parameters and spleen cell peroxidative activity. Histologic examination of these animals revealed no evidence of mycoplasma infection. The observed differences between exposed and control animals of the first experiment appear to have resulted from subclinical respiratory mycoplasmosis rather than exposure to RFR.
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PMID:Effects of 20-MHz radiofrequency radiation on rat hematology, splenic function, and serum chemistry. 402 74

The cytoplasmic membrane of Mycoplasma laidlawii strain B is solubilized by anionic and nonionic detergents, succinylation, phospholipase A, alkaline phosphatase, trypsin, and chymotrypsin. Cationic detergents are without effect, as are chelating agents, even in the presence of high concentrations of monovalent cation. The detergent-solubilized membrane exhibits one peak in the analytical ultracentrifuge, but the sedimentation coefficient is dependent upon concentration of detergent. Simple dialysis does not remove all of the sodium dodecylsulfate except from lipid-depleted membrane particles. Membranes bind sodium dodecylsulfate but acetone powders of membranes do not. Sulfated alcohols with chain lengths of C(14) and C(16) are more tightly bound than dodecylsulfate. A constant amount of di- and trivalent cation is bound by the membrane upon aggregation. Only a portion of this cation is removable with chelating agents. No chelating agent is bound by these aggregates. A portion of the lipid-depleted membrane particles is solubilized by negatively charged lipids and detergents, giving rise to aggregates in the presence of divalent cation. Fractionations of detergent-solubilized membranes by preparative gel electrophoresis and ammonium sulfate were inconclusive. Density gradient centrifugation of succinylated membranes yielded at least five fractions which exhibited homogeneity by ultracentrifugation. Analytical gel electrophoresis of these fractions demonstrated heterogeneity. The composition of these five fractions suggested separation of protein from lipid.
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PMID:Observations on membranes of Mycoplasma laidlawii strain B. 536 Dec 9

We developed a microtitre hybridization assay for the detection of polymerase chain reaction (PCR) amplified sequences. For this, cloned Mycoplasma pneumoniae DNA containing a sequence complementary to the PCR products is first covalently bound to microtitre wells. These coated microplates can be stored for several months. Then, an aliquot of the PCR product, labelled with digoxigenin-dUTP during its synthesis is hybridized to the immobilized DNA. The use of a rapid hybridization buffer makes this step very short (5 min). Finally, the hybridization signal is detected by an anti-digoxigenin antibody conjugated with alkaline phosphatase. Compared to Southern or other microplate hybridization techniques, this method is cheaper, involved fewer steps and allows easy handling of a large number of samples. This method was used for detection of M. pneumoniae in a series of clinical specimens.
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PMID:Improved microplate immunoenzymatic assay of PCR products for rapid detection of Mycoplasma pneumoniae. 776 Aug 56

Enzyme-linked immunosorbent assay (ELISA) for IgA, IgG and IgM was evaluated with sera from 50 adult patients with pneumonia, selected on the basis of a positive complement fixation (CF) test for diagnosis of Mycoplasma pneumoniae infection and with sera from 105 healthy blood donors. The ELISA antigen for IgG and IgA was a sonicated suspension of M. pneumoniae solubilised by deoxycholate. For the IgM assay, the same antigen was directly conjugated to alkaline phosphatase and used in a mu-capture format. ELISA gave positive results with high or rising titres for one or several antibody classes in 47 (94%) patients. In two of the three ELISA-negative cases, the diagnosis of M. pneumoniae infection indicated by the CF test seemed unlikely on clinical grounds. Specific IgA antibodies was developed more regularly and more rapidly than IgM. IgA titres also started to decrease earlier than IgM or the late-peaking IgG response. Thus, the determination of IgA antibodies was found to be valuable for the early diagnosis of M. pneumoniae infection. The study also demonstrated that the determination of all three antibody classes is necessary to obtain an optimal level of serodiagnosis.
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PMID:The role of IgA determination by ELISA in the early serodiagnosis of Mycoplasma pneumoniae infection, in relation to IgG and mu-capture IgM methods. 815 81

Chlamydia pneumoniae infections have earlier been described as mycoplasma-like illnesses in young people, and also appear to be associated with community-acquired pneumonia in adults. In this retrospective study, 12.2% (23/188) of patients with pneumonia who required hospitalization during the 3 years 1985-87 had serological evidence of recent C. pneumoniae infection. Many of these patients had symptoms similar to ornithosis. The most interesting finding was that half of the patients with a 4-fold IgG antibody titre rise to C. pneumoniae also had an increased alkaline phosphatase concentration.
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PMID:Chlamydia pneumoniae in Swedish patients. 824 41

Mycoplasma pneumoniae is the causative agent of primary atypical pneumoniae, and two distinct groups (I and II) have been established. Serological tests are relatively insensitive and the diagnosis by culture is time-consuming. This study was therefore undertaken to detect and to identify M. pneumoniae on culture media and in throat swab specimens by using polymerase chain reaction (PCR) and hybridization probes conjugated to alkaline phosphatase (Alp). Primer pairs were selected for amplification of DNA fragments in the C to D, F, G and I to J regions of the M. pneumoniae cytadhesin P1 genes. Amplified DNA fragments were visualized by staining with ethidium bromide after 2% agarose gel electrophoresis and by Southern hybridization with Alp-labeled probes. No amplification of the P1 genes was seen with any of five related Mycoplasma species, the others from M. pneumoniae. In all of 30 clinical isolates on PPLO medium, M. pneumoniae was detected with the F and G primer pairs, giving 100% of sensitivity. Of 69 throat swab specimens, 25 were positive with the F primer pairs, and 23 positive with the Gen Probe test. From these results, we conclude that the PCR with F or G primer pairs can be adapted as a practical method for the rapid diagnosis of M. pneumoniae infections.
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PMID:[Detection of Mycoplasma pneumoniae by using polymerase chain reaction and nonradioactive DNA probes]. 856 36

A polymerase-chain-reaction-based detection system for Mycoplasma fermentans was established. The highly conserved tuf gene, which encodes elongation factor Tu of prokaryotes, served as target sequence for the PCR. With two PCR oligodeoxynucleotides, which were selected from M. fermentans specific sequences of the tuf gene, we amplified a 850 base pair DNA fragment. Via the biotin-moiety of one primer the PCR fragments were immobilized on streptavidin-coated microtitre plates. After alkaline denaturation a digoxigenin-labelled M. fermentans specific DNA probe was hybridized to the single stranded immobilized PCR fragment. Detection was performed by addition of an alkaline phosphatase conjugated anti-digoxigenin antibody. 4-methyl-umbelliferyl-phosphate was used as a fluorogenic substrate. Amplification of 10 fg chromosomal target DNA was detected by this 'DNA enzyme immuno assay (DEIA)' technique, corresponding to seven genome copies. Our study supports the presumption that the tuf gene proves to be a suitable target sequence for the PCR based detection of any bacterial species. Furthermore, hybridization of PCR fragments with radio-labelled DNA probes should no longer be necessary, because a very sensitive non-radioactive test system can easily be established with the 'DEIA' technique.
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PMID:Development of an amplification and hybridization assay for the specific and sensitive detection of Mycoplasma fermentans DNA. 868 79

Short nucleotides directly labelled to alkaline phosphatase (SNAP probes) are an interesting alternative to digoxigenin-labelled probes (DIG probes), because they reduce the number of steps necessary in dot blots for the detection of DNA or amplificate. This study examined the questions whether a SNAP probe might not only save time, but also increase the sensitivity of another PCR-based DNA probe test using a digoxigenin probe. Amplificates obtained by multispecies polymerase chain reaction (PCR), with either purified genomic DNA or DNA extracted from tracheal swabs taken in chicken flocks, were detected by both methods. The results for the clinical specimens were compared to culture. Under stringent conditions, the specificity and sensitivity obtained with the SNAP probe were comparable to the results obtained with the DIG probe. The quantities 10 fg (SNAP probe) and 100 fg (DIG probe) of purified Mycoplasma synoviae DNA were detected after amplification, but more positive clinical specimens were detected with the DIG probe. Under non-stringent conditions sensitivity with purified DNA did no change, but the coloration of the dots improved markedly, and more positive specimens could be detected with the SNAP probe than with the DIG probe, truly positives as confirmed by culture. Because cross-reaction with Mycoplasma gallisepticum and Mycoplasma imitans, two species with DNA that was also recognized by the multispecies primers, occurred under non-stringent conditions, it was concluded that, to take the full advantage of SNAP probes, their use in combination with species-specific primer pairs is recommended. PCR as a method for mycoplasma detection is however, always accompanied with serological and cultural methods. When a M. synoviae mono-infection is likely by serological results, non-stringent dot blot conditions and use of the SNAP probe will ease and improve the detection of mycoplasma.
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PMID:Use of an alkaline phosphatase-labelled probe for the detection of Mycoplasma synoviae in chickens. 1078 Jan 70

This study established a modified alkaline phosphatase-labelled avidin-biotin-complex (ABC-AP) method for diagnosis of mouse hepatitis virus (MHV) and Mycoplasma pulmonis infection from formalin-fixed, paraffin wax-embedded sections, murine antibody-positive serum being used as the primary reagent. With this method, MHV antigen in cAnNCrj.Cg-Foxn1(nu)/Foxn1(nu) mice and M. pulmonis antigen in Wistar rats were immunolabelled in tissue sections. MHV antigen was clearly detected in samples of liver, stomach, caecal and colonic mucosa, and spleen. M. pulmonis antigen was demonstrated on the luminal surface of bronchiolar epithelial cells. This method may prove useful in diagnosis when commercial antisera are unavailable or when immunosuppression prevents serological diagnosis.
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PMID:Immunohistochemical diagnosis of mouse hepatitis virus and mycoplasma pulmonis infection with murine antiserum. 1527 61

Babesia felis, originally identified in wild cats in the Sudan, was subsequently found to cause clinical disease in domestic cats. Although babesiosis in domestic cats has been reported sporadically from various countries, as a significant disease it appears to be a distinctly South African phenomenon. Apart from an inland focus, feline babesiosis is reported regularly only from coastal regions. The infection is assumed to be tick-borne, but the vector has not been identified. Feline babesiosis tends to be an afebrile, chronic, low-grade disease. The most frequently reported complaints by owners are anorexia and lethargy. The main clinical findings are anemia, depression, and occasionally icterus. Concurrent infections (e.g., Mycoplasma haemofelis, FeLV, FIV) may contribute to the clinical picture. Laboratory findings commonly include regenerative anemia, elevation of alanine transaminase (but not alkaline phosphatase) and total bilirubin concentrations, and a variety of electrolyte disturbances. Secondary immune-mediated hemolytic anemia can be seen occasionally. Drugs effective against other Babesia species give variable and questionable results. The drug of choice is primaquine phosphate, which effects a clinical cure but does not sterilize the infection. Repeated or chronic therapy may be required.
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PMID:Feline babesiosis in South Africa: a review. 1560 90


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