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PubMed2000

The prevalence of antibodies to anionic phospholipids in patients with the primary antiphospholipid syndrome, systemic lupus erythematosus and their relatives and spouses.

OBJECTIVES: Antiphospholipid antibodies (aPL) have been associated with syndromes involving thrombosis, fetal loss and thrombocytopenia. Genetic and environmental conditions are among the factors attributed to the cause of autoimmune diseases such as the antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE). The aim of this study was to determine whether these factors determine the prevalence of aPL. METHODS: Three groups of patients were tested for the presence of IgG, IgM and IgA anticardiolipin (aCL), antiphosphatidylinositol (aPI), antiphosphatidylglycerol (aPG) and antiphosphatidylserine (aPS) antibodies: (i) patients with primary APS (PAPS); (ii) patients with SLE and secondary APS; and (iii) patients with SLE without APS. First-degree relatives and spouses of patients with SLE/APS were also tested for circulating aPL. RESULTS: IgG aPL were particularly prevalent in patients with PAPS. IgG aPI and aCL were more prevalent in patients with PAPS than the IgM equivalents (P < 0.0001). Notably, none of the patients with PAPS had IgA aPL. A significantly higher number of relatives of patients with SLE/APS possessed IgG aPL than the normal controls. Except for aPG (P < 0.03), the prevalence of these antibodies in the relatives was not significantly different from patients with SLE/APS. The relatives also had significantly higher prevalence of IgG aPI, aPS and aCL antibodies than IgM aPL antibodies. In contrast, the prevalence of IgG aPL in the spouses was no different than in the healthy controls. CONCLUSIONS: Genetic factors, shared by patients and their relatives, seem to have some effect on the prevalence of aPL in the subjects studied, while environmental factors shared by spouses appear to have no influence.

Adult↗
PubMedDate not supplied

A study of functional viability and metabolic degeneration of human skin stored at 4 degrees C.

We sought to ascertain whether an in vitro assay could be as reliable as an in vivo assay in determining the viability of human skin stored at 4 degrees C. Allografts from six human donors were stored in RPMI 1640 tissue culture medium at 4 degrees C. At fixed intervals during the storage period, all skin specimens were tested concurrently by two different viability assays: (1) transplantation onto surgically created defects on nude mice, and (2) intracellular enzyme activity with use of a 4-hour semiquantitative micromethod system activity (API ZYM; Biomerieux Vitek Inc., Hazelwood, Mo.). Human graft survival on the nude mice was 100% for the first 15 days of storage, and then declined to 50% on storage day 30. The API ZYM assay showed a comparable progressive decrease in enzyme activity over skin storage time. The API ZYM assay is a simple, rapid system that produces reproducible results and is cost-effective when compared to the biologic model.

Animals↗
PubMed2004

An opportunity for prevention: prevalence, incidence, and sexual risk for HIV among young Asian and Pacific Islander men who have sex with men, San Francisco.

OBJECTIVES: We investigated HIV prevalence and incidence and related risk factors among young Asian and Pacific Islander (API) men who have sex with men (MSM). DESIGN: We conducted a cross-sectional survey. METHODS: A venue-based sample of 496 young API MSM in San Francisco completed face-to-face questionnaires and received HIV counseling and testing. RESULTS: HIV prevalence was 2.6% and annualized HIV incidence was 1.8% per year. In multivariate analysis, being American-born, having 51+ lifetime sex partners, and having attended a "circuit party" (multiday large MSM gatherings) were associated with HIV infection. Forty-seven percent of the sample reported unprotected anal intercourse in the past 6 months. CONCLUSIONS: The current levels of HIV prevalence, HIV incidence, and sexual risk behavior suggest an emerging HIV epidemic among young API MSM in San Francisco.

Adolescent↗
PubMed2003

Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum.

BACKGROUND: Protection against complement activation may reduce acute islet damage in pig-to-human islet xenotransplantation. Expression of the human complement regulatory proteins decay-accelerating factor (DAF, CD55) or CD59 was induced on intact adult porcine islets (APIs) by adenoviral transduction. The functional capacity of the transgenes was examined in vitro after exposure to fresh human serum. METHODS: Intact APIs were transduced with adenoviral vectors Ad.hDAF or Ad.hCD59 or a control vector. After 3 days, the islets were trypsin dissociated to a single-cell suspension. A cytotoxicity assay was performed in which the islet cells were incubated with human complement active AB serum. Flow cytometry and immunohistochemistry were used to evaluate transgene expression. RESULTS: APIs could be transduced to express hDAF or hCD59. Flow cytometry analysis of islet single cells revealed that only a fraction of the cells expressed the transgene; immunohistochemical staining of transduced islets demonstrated that mainly cells located in the periphery of the islets were expressing the protein. Cells from nontransduced islets or islets expressing the control protein were sensitive to lysis in human sera (66+/-4.0% and 73+/-3.7% cytotoxicity, respectively). Single cells from islets transduced with hDAF and hCD59 were partially protected from lysis. Islet cells expressing hCD59 were slightly less sensitive to lysis (33+/-3.3%) than cells expressing hDAF (45+/-3.5%). CONCLUSIONS: These data show that intact pig islets can be transduced to express human regulators of complement activation on the surface and that pig islet cells expressing hDAF or hCD59 are less sensitive to complement-mediated lysis.

Adenoviridae↗
PubMed2003

Immunosuppression with FTY720 and cyclosporine A inhibits rejection of adult porcine islet xenografts in rats.

BACKGROUND: Our aim was to evaluate the effect of FTY720 in discordant islet xenotransplantation. METHODS: Fetal porcine islet-like cell clusters (ICCs) were transplanted into normoglycemic rats that were either left untreated or treated with FTY720 only, with FTY720 plus cyclosporine A (CsA) or with CsA only. Twelve or 24 days after transplantation, graft morphology was evaluated immunohistochemically. Furthermore, adult porcine islets (APIs) were transplanted into diabetic rats immunosuppressed with FTY720 plus CsA. Blood glucose and porcine C-peptide levels were monitored. RESULTS: In untreated rats, the ICC xenografts were completely rejected after 12 days. Treatment with CsA had only a marginal effect on the rejection. In animals given FTY720, only the number of infiltrating cells was somewhat reduced. However, at 12 days, no intact ICCs remained. Immunosuppression with FTY720 plus CsA had a marked inhibitory effect on islet xenograft rejection and plentiful morphologically intact ICCs remained. Twelve days after transplantation, only occasional macrophages and T cells could be detected. At 24 days after transplantation, the findings were similar. Furthermore, diabetic rats transplanted with APIs and immunosuppressed with FTY720 plus CsA remained normoglycemic for 53.0+/-15.8 days. In fact, one animal remained normoglycemic for more than 100 days. Serum levels of porcine C-peptide remained at levels similar to those for human C-peptide in healthy individuals. CONCLUSIONS: Immunosuppression with FTY720 plus CsA inhibited almost all morphological signs of pig-to-rat islet xenograft rejection for up to 24 days after transplantation. Diabetic rats transplanted with APIs and immunosuppressed with FTY720 plus CsA remained normoglycemic for 53.0+/-15.8 days.

Animals↗
PubMed1976

An iridovirus from bees.

An iridovirus, Apis iridescent virus (AIV), isolated from sick adult specimens of Apis cerana (Hymenoptera) from Kashmir, closely resembles iridescent viruses from Tipula and Sericesthis spp. (TIV and SIV). However, AIV is only distantly related serologically to TIV and SIV and is even more remotely related to several other similar viruses that were tested in tube precipitation tests with intact particles. AIV multiplies in Apis mellifera, forming cytoplasmic iridescent crystalline aggregates in several tissues, but unlike all the other iridoviruses tested, it failed to multiply in Galleria mellonella.

Animals↗
PubMed2004

Anoxybacillus contaminans sp. nov. and Bacillus gelatini sp. nov., isolated from contaminated gelatin batches.

Aerobic, endospore-forming bacteria that are attributed to the genus Bacillus or related genera constitute a hazard to the quality of gelatin. During repetitive extragenic palindromic DNA (rep)-PCR screening of gelatin isolates, a group of five isolates (group 1) and a group of 66 isolates (group 2) that did not match any pattern in our database were found. On the basis of 16S rDNA sequence analysis, representative strains of the different rep-PCR fingerprint types of group 1 were shown to be related most closely to Anoxybacillus species, but with sequence similarity of <97 %. Likewise, representative strains of group 2 were shown to be related most closely to Bacillus species, with 16S rDNA sequence similarity of <97 %. DNA-DNA reassociation values of isolates that displayed the most divergent rep-PCR profiles revealed that strains within each group belonged to a single species, according to recommendations for species delineation. A mean fatty acid profile could be calculated for each group. Isolates within a single group had similar patterns of results in API and other phenotypic tests; no correlation of patterns of results with rep-PCR groups was seen. Physiological characterization of group 1 isolates allows their distinction from other Anoxybacillus species. Despite the weak reaction of group 2 isolates in API tests, physiological characterization allows distinction between Bacillus species that react weakly in API tests. Two novel species are therefore proposed, with the names Anoxybacillus contaminans sp. nov. (type strain, LMG 21881(T)=DSM 15866(T)) and Bacillus gelatini sp. nov. (type strain, LMG 21880(T)=DSM 15865(T)).

Bacillaceae↗
PubMed2005

Evaluation of commercial kits for the identification of Neisseria gonorrhoeae.

Eight identification methods were evaluated against 100 isolates of Neisseria gonorrhoeae and 21 non-gonococcal Neisseria strains. The methods examined included four commercial biochemical kits, API NH, RapID NH, Gonochek II and Neisseria Preformed Enzyme Test (PET), three immunological kits, Phadebact Monoclonal GC test, GonoGen II and MicroTrak, and one in-house carbohydrate-utilization method, cystine trypticase agar (CTA) sugars. The percentage of isolates unambiguously identified as N. gonorrhoeae by each of the methods was as follows: API NH, 66 %; RapID NH, 64 %; GonoChek II, 66 %; Neisseria PET, 66 %; Phadebact Monoclonal GC OMNI test, 99 %; GonoGen II, 100 %; MicroTrak, 100 %; and CTA sugars, 96 %. The low sensitivity of the biochemical kits for the identification of N. gonorrhoeae was due to a lack of the enzyme proline iminopeptidase (Pip) in 34 % of the isolates examined. All the biochemical kits utilized the presence of this enzyme as a marker for N. gonorrhoeae. The Phadebact Monoclonal GC kit, GonoGen II, MicroTrak, CTA sugars and the API NH kit all exhibited high specificity, but non-gonococcal Neisseria were misidentified as N. gonorrhoeae using RapID NH (two strains), Gonochek II (11 strains) and Neisseria PET (11 strains). Whilst the isolates examined in this study may not be truly representative, they do indicate that N. gonorrhoeae isolates lacking the enzyme Pip can give anomalous results when using commercially available biochemical tests and that some non-pathogenic Neisseria species are still being misidentified using some biochemical kits. This further reinforces the recommendation that any dubious biochemical result should be confirmed with an immunological test.

Bacterial Typing Techniques↗
PubMed2006

Evolution of the complementary sex-determination gene of honey bees: balancing selection and trans-species polymorphisms.

The mechanism of sex determination varies substantively among evolutionary lineages. One important mode of genetic sex determination is haplodiploidy, which is used by approximately 20% of all animal species, including >200,000 species of the entire insect order Hymenoptera. In the honey bee Apis mellifera, a hymenopteran model organism, females are heterozygous at the csd (complementary sex determination) locus, whereas males are hemizygous (from unfertilized eggs). Fertilized homozygotes develop into sterile males that are eaten before maturity. Because homozygotes have zero fitness and because common alleles are more likely than rare ones to form homozygotes, csd should be subject to strong overdominant selection and negative frequency-dependent selection. Under these selective forces, together known as balancing selection, csd is expected to exhibit a high degree of intraspecific polymorphism, with long-lived alleles that may be even older than the species. Here we sequence the csd genes as well as randomly selected neutral genomic regions from individuals of three closely related species, A. mellifera, Apis cerana, and Apis dorsata. The polymorphic level is approximately seven times higher in csd than in the neutral regions. Gene genealogies reveal trans-species polymorphisms at csd but not at any neutral regions. Consistent with the prediction of rare-allele advantage, nonsynonymous mutations are found to be positively selected in csd only in early stages after their appearances. Surprisingly, three different hypervariable repetitive regions in csd are present in the three species, suggesting variable mechanisms underlying allelic specificities. Our results provide a definitive demonstration of balancing selection acting at the honey bee csd gene, offer insights into the molecular determinants of csd allelic specificities, and help avoid homozygosity in bee breeding.

Alleles↗
PubMed2000

The 1:1 adduct of Kemp's triacid with 2-aminopyridine

The crystal structure determination of the molecular proton-transfer adduct of Kemp's triacid (cis-cis-1,3,5-trimethylcyclohexane-1,3, 5-tricarboxylic acid, KTA) with 2-aminopyridine (2-APY), namely 2-aminopyridinium 3,5-dicarboxy-1,3, 5-trimethylcyclohexanecarboxylate, 2-APY(+).KTA(-) or C(5)H(7)N(2)(+).C(12)H(17)O(6)(-), has revealed a centrosymmetric hydrogen-bonded cyclic KTA homodimer repeating unit [O.O 2.524 (4) A] linked into a polymer structure through the pyridinium and amino groups of the 2-APY molecule [O.N 2.736 (4), 2.989 (4) and 2.999 (4) A].

Journal Article↗
PubMedDate not supplied

Adjuvant chemotherapy with cisplatin, ifosfamide, and doxorubicin followed by radiotherapy in localized uterine sarcomas: results of a case-control study with radiotherapy alone.

UNLABELLED: Uterine sarcoma is a poor prognosis disease, with a high risk of metastatic relapse. We conducted a study of adjuvant chemotherapy with cisplatin, ifosfamide, and doxorubicin followed by radiotherapy (n=18). The results were then compared in a matched case-controlled study to radiotherapy alone (n=16) or no therapy at all (n=2). Chemotherapy consisted in three cycles of adriamyein-platinum-ifosfamide (API) (doxorubicin 60 mg /m2 on day 1; cisplatin 100 mg /m2 on day 2; ifosfamide 5 g /m2 on day 1+mesna 5 g /m2 on day 1+granulocyte colony-stimulating factor; q 3 weeks). Drug doses were reduced (20% for ifosfamide and cisplatin) four times (four patients) due to hematologic toxicity. Compared to a case-control study of adjuvant radiotherapy alone, results were not decreased by the addition of a toxic chemotherapy. CONCLUSION: Adjuvant API chemotherapy followed by radiotherapy is a feasible protocol; a multicenter phase III study comparing radiotherapy alone versus API chemotherapy followed by radiotherapy just began in France.

Adult↗
PubMed1986

Numerical taxonomy of alpha-amylase producing Bacillus species.

A total of 134 alpha-amylase producing Bacillus isolates and 21 reference strains were divided into 12 groups according to their similarities (% SSM). Phenotypic characteristics determined by the API 20E and API 50CHB galleries, other biochemical tests and morphological characteristics were used for the numerical analysis. The API Computer Service identified 45% of the isolates. The amylase yields of 16 alpha-amylase hyperproducing (AHP) isolates were compared with those of seven amylolytic reference and type strains. The AHP isolates were related to Bacillus subtilis, B. licheniformis and 'B. amyloliquefaciens'.

Bacillus↗
PubMed1998

Characterization of coagulase-negative staphylococci by primer-specific polymerase chain reaction and ribotyping.

OBJECTIVE: To analyze, by primer-specific polymerase chain reaction (PCR) and ribotyping, coagulase-negative staphylococci (CNS). METHODS: Forty-five clinical isolates of CNS were identified by the API ID32 STAPH system and ribotyping. Additionally, primer-specific PCR was evaluated for identification of clinical strains of Staphylococcus epidermidis. RESULTS: Forty-five isolates of CNS from neonates with nosocomial bacteremia were studied. The results of the S. epidermidis-specific PCR were compared with those obtained using ribotyping and the API ID32 STAPH system. Excellent congruence was found between primer-specific PCR and ribotyping. Primer-specific PCR proved to be a fast and reliable method for the identification of S. epidermidis strains. According to the primer-specific PCR and ribotyping analysis, a few CNS isolates were found to be incorrectly identified by the API ID32 STAPH system. CONCLUSIONS: Primer-specific PCR is a fast and reliable method for the identification of S. epidermidis. Primer-specific PCR in combination with ribotyping is a promising approach for studying the epidemiology of S. epidermidis and other CNS species in hospital.

Journal Article↗
PubMed1996

Comparison of a commercial biochemical kit and an oligonucleotide probe for identification of environmental isolates of Vibrio vulnificus.

Methods for the identification and isolation of environmental isolates of Vibrio vulnificus were evaluated. Alkaline peptone water supplemented with polymyxin B and colistin-polymyxin B-cellobiose agar were employed for the isolation of suspected V. vulnificus from water, sediment and shellfish samples. When comparing the identification of putative V. vulnificus obtained with the API 20E assay and an oligonucleotide probe, 29 API 20E profiles were obtained with only four profiles (representing 20 isolates) reaching the identification threshold of V. vulnificus among a total of 66 isolates hybridizing with the probe. The results indicated that, compared with colony hybridization, the API 20E assay was not adequate for the identification of environmental isolates of V. vulnificus.

Bacterial Typing Techniques↗
PubMed2004

Screening of lactic acid bacteria from fermented vegetables by carbohydrate profiling and PCR-ELISA.

AIMS: The aim of this study was to identify potential souring agents, isolated from fermented plant material, by API 50 CHL assay and a molecular method based on polymerase chain reaction and colorimetric hybridization (PCR-ELISA). METHODS AND RESULTS: Forty-two strains of lactic acid bacteria derived from plant material were screened by taking advantage of API 50 CHL and PCR-ELISA. Oligonucleotide probes used for hybridization in PCR-ELISA were specific for lactobacilli, the Leuconostoc family, Lactobacillus pentosus/plantarum and Lactobacillus brevis. The hybrides were detected by a colour-developing reaction. Bacteria isolated from fermented cucumbers were identified as Lact. plantarum-related (Lact. plantarum and Lact. pentosus) and Leuconostoc species. Most of the strains isolated from sauerkraut were identified as Lact. pentosus/plantarum. CONCLUSIONS: Complementary results were obtained in the identification of bacterial strains, isolated from fermented cucumbers and sauerkraut, by API 50 CHL and PCR-ELISA. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-ELISA proved to be suitable for the screening of large numbers of bacterial isolates from fermented vegetables. This will be useful for the identification of strains suitable for the design of starter cultures for the fermentation of plant material.

Carbohydrates↗
PubMed1996

Salivary peroxidase activity in whole saliva of patients with insulin-dependent (type-1) diabetes mellitus.

In this study, salivary peroxidase activity was measured in a group of 10 patients with insulin-dependent (type I) diabetes mellitus (IDDM) who had a tendency towards periodontitis. In healthy subjects (N : 10), mean salivary peroxidase activity was 0.0025 +/- 0.001 IU/ml, while in this group of type I diabetic patients it was 0.051 +/- 0.015 IU/ml, a significantly higher level (p < 0.001). Approximal plaque index (API), modified sulcus bleeding index (mod SBI) and pocket depths were assessed clinically. The values for mod SBI and API were 60% and 68% respectively for the diabetic patients while for the control group mod SBI was measured 0.0% and the value for API was 10.67% (p < 0.001). The administration of this simple and practical test may provide an early marker of a tendency towards periodontitis in IDDM patients.

Adolescent↗
PubMed1994

Identification of coryneform and other gram-positive rods with several methods.

The identification of 202 isolates of aerobically growing Gram-positive rods from clinical material was attempted by using a combination of "traditional" morphological and biochemical tests (Hollis & Weaver (20)) plus patterns of cellular and metabolic fatty acids. This system served as the "gold standard" for three others, i.e. API Coryne (Rapid Coryne), MIDI TSBA and MIDI CLIN Aerobic. In addition, several growth, biochemical and susceptibility tests (growth on cystine-tellurite blood agar, DNase, hippurate and starch hydrolysis, methanethiol formation, API ZYM, CAMP reaction, susceptibility to O/129 and to six antimicrobials) were done in order to check their usefulness for the identification of this group of bacteria. Our system, with the help of chemotaxonomic tests (m-DAP and mycolic acids), was able to identify 154/202 (76%) of the isolates by species and an additional 41/202 (21%) by genus only; 7 (3%) could not be identified. The API Coryne system identified to species or genus level 140/195 isolates (72%). Corresponding figures for the MIDI TSBA and CLIN systems were 63/195 (32%) and 88/195 (45%); further details of species and genus identification are presented in the text. The main drawback of the commercial systems is the extent and probably the numerical depth of the data base. We recommend the use of our multisystem approach for the identification of Gram-positive rods until commercial systems are based on a broader and numerically more extensive data base. The additional tests did not prove species- or genus-specific.

Culture Media↗
PubMed1994

Comparison of genotypic and phenotypic methods for species identification of coagulase-negative staphylococcal isolates from blood cultures.

Coagulase-negative staphylococci are often isolated from blood cultures. Simple methods are needed for correct identification of those species most frequently found. In this study, PCR methods were developed for the identification of S. epidermidis and S. haemolyticus, based on DNA sequences in their 16S rDNA. The results obtained by these methods were compared with those obtained using a number of phenotypic methods, including two commercial kits API Staph and Staphzyme. Fifteen type collection strains and 133 blood culture isolates were tested. The sensitivity of the PCR for identification of S. epidermidis was 99% compared with API Staph, but the specificity was lower, 94%, because of positive results also for S. capitis. The results by the PCR for identification of S. haemolyticus correlated closely with the Staphzyme results, 13 isolates being identified by Staphzyme and 16 by the PCR. API Staph, however, identified only four clinical isolates as S. haemolyticus, probably too few. Among the individual phenotypic tests performed, a trehalose-mannitol agar method and a desferrioxamine disc diffusion test for the identification of S. epidermidis were found to be very accurate. Anaerobic growth after overnight incubation could be used to distinguish S. epidermidis from S. hominis. The conclusion is that a majority of all Gram-positive, catalase-positive and coagulase-negative blood culture isolates can be typed as regards species level using only a few genotypic and/or phenotypic tests.

Bacterial Typing Techniques↗