Purified, labelled 16 S amplicons were then hybridized to the pri

Purified, labelled 16 S amplicons were then hybridized to the printed HOMIM slides at 55°C for 16 h. Hybridized slides were washed and dried and Cy3 fluorescence was detected using the GenePix 4000B microarray scanner (Axon) with photomultiplier settings (PMT) of 650 and wavelength of 532 nm. Analysis of HOMIM data Analysis of HOMIM data was performed as previously described [42, 43]. Briefly, hybridization spot intensities were converted to one of the 6 integer signal levels ranging from 0 to 5, with 0 representing undetectable (above background) and 5 being the maximal intensity among all the profiles being compared.

The number of bacterial species (Species Score) present in each sample was determined by summation of all learn more probes with detectable signal (integer score ≥ 1), and a qualitative representation of the total bacteria (Bacterial Load) in each sample was estimated by summation of all integer scores. Correlations between “Species Score” and “Bacterial Load” were analyzed using Spearman rank correlation coefficient. Correlations between clinical

Fer-1 parameters (viral loads, CD4+ T cell counts) and a gain or loss of oral bacteria were identified by Spearman rank correlation coefficient analysis. Wilcoxon rank-sum tests were utilized to determine if increases or decreases in individual bacterial species in HIV patient groups were statistically significant compared to healthy HIV- controls. The HOMIM data utilized in the study has been deposited in the Gene Expression Omnibus microarray database (Accession#: GBA3 GSE38908). Acknowledgements The authors would like to thank the clinicians and staff at the Center for AIDS Research and Education (CARES) Clinic in Sacramento, CA for their help in scheduling patient appointments and collecting samples. This study was funded through a pilot grant from the California Research Center for

the Biology of HIV in Minorities (CRCBHM). Statistical support was made possible through funding (UL1 RR024146) from the National Center for Research Resources (NCRR). References 1. McCune JM: The dynamics of CD4+ T-cell depletion in HIV disease. Nature 2001,410(6831):974–979.PubMedCrossRef 2. Egusa H, Soysa NS, Ellepola AN, Yatani H, Samaranayake LP: Oral candidosis in HIV-infected patients. Curr HIV Res 2008,6(6):485–499.PubMedCrossRef 3. Hazenberg MD, Hamann D, selleckchem Schuitemaker H, Miedema F: T cell depletion in HIV-1 infection: how CD4+ T cells go out of stock. Nat Immunol 2000,1(4):285–289.PubMedCrossRef 4. Reznik DA: Oral manifestations of HIV disease. Top HIV Med 2005,13(5):143–148.PubMed 5. Myers TA, Leigh JE, Arribas AR, Hager S, Clark R, Lilly E, Fidel PL: Immunohistochemical evaluation of T cells in oral lesions from human immunodeficiency virus-positive persons with oropharyngeal candidiasis. Infect Immun 2003,71(2):956–963.PubMedCrossRef 6.

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