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Biomedical subjects

P C Tsang

Publications and source records attributed to P C Tsang.

13 recordsLinked to original sources

Heterogeneity in antifungal susceptibility of clones of Candida albicans isolated on single and sequential visits from a HIV-infected southern Chinese cohort.

The increased frequency and severity of candidal infections in human immunodeficiency virus (HIV)-infected individuals has prompted the wide use of antifungals, such as amphotericin B, ketoconazole, and fluconazole, resulting in the emergence of drug-resistant strains of Candida albicans. To study this phenomenon in an ethnic Chinese cohort, we isolated multiple colonies of Candida from the oral cavities of 16 HIV-infected patients on single and subsequent sequential visits over a period of 12 months. Ten of the 16 patients had sporadic episodes of oropharyngeal candidiasis (Group A), while the remainder were asymptomatic with respect to this condition (Group B). Oral rinses were collected and immediately processed in the laboratory for the isolation of C. albicans in a standard manner. A total of 433 C. albicans isolates were tested for their susceptibility to amphotericin B, ketoconazole and fluconazole by an agar diffusion method using the commercially available E-test. All tested isolates demonstrated variable susceptibility to amphotericin B, ketoconazole and fluconazole. The minimum inhibitory concentration (MIC) of the isolates for amphotericin B, ketoconazole and fluconazole ranged from <0.002-1.5 microg/ml, <0.002-4.0 microg/ml and <0.016-32 microg/ml, respectively. Sequential isolates of a few patients demonstrated variable susceptibility to all the antifungals, and no discernible MIC pattern emerged either in group A or B over time. Interestingly, significant variation in antifungal susceptibility was also noted in isolates obtained from the same patient on a single visit. Sequential yeast isolates in 9 of 16 patients (56%) demonstrated significant differences in MIC within and between visits for both amphotericin B and ketoconazole, while a lower percentage--44%(7/16)--exhibited this trait for fluconazole. Our study demonstrates the diversity in antifungal susceptibility in either commensal or "infective" oral strains of C. albicans in HIV disease, and shows the need for vigilance for the emergence of resistant strains, and for frequent antifungal susceptibility studies.

AIDS-Related Opportunistic Infections↗

Oral health status, oral microflora, and non-surgical periodontal treatment of renal transplant patients receiving cyclosporin A and FK506.

OBJECTIVES: To determine the oral health status, oral microflora and the effect of non-surgical periodontal treatment on gingival overgrowth of renal allograft recipients receiving either cyclosporin A (CsA) or FK506 (Tacrolimus) as an immunosuppressant. MATERIALS AND METHODS: A total of 47 patients receiving CsA (mean age 43.1 years) and 10 receiving FK506 (mean age 40.1 years) were included in the study. Stone casts were taken for measurement of gingival overgrowth. An oral rinse technique was used to investigate the prevalence of yeasts, and aerobic and facultatively anaerobic Gram-negative rods (AGNR). RESULTS: The CsA and FK506 patients exhibited a Gingival Overgrowth Index (GOI) of 45.2%, and 25.1%, respectively (p < 0.05). The CsA patients had a GOI of 15.2% after one year of non-surgical periodontal treatment. The difference between pre- and postoperative gingival overgrowth indices was significant (p < 0.0001). Candida albicans and Klebsiella pneumoniae were the most notable yeast and AGRN found. CONCLUSIONS: Renal transplant patients, being immunocompromised, constitute a high-risk group for gingival overgrowth. However, the FK506 regime appeared to ameliorate this effect, compared with CsA. Non-surgical periodontal treatment was effective in reducing established gingival overgrowth in both CsA and FK506 patients (p < 0.05). Adequate pre- and post-transplant oral health care is recommended, for these patients, irrespective of the drug regime.

Adult↗

Oral manifestations of HIV infection in a group of predominantly ethnic Chinese.

A total of 32 HIV-infected, predominantly ethnic Chinese individuals from Hong Kong were examined for oral mucosal lesions over a period of 1 year. The commonest oral lesion found was minor aphthous ulceration (27.4%), while xerostomia (17.8%), ulceration NOS (not otherwise specified; 12.3%), hairy leukoplakia (11.0%) and erythematous candidiasis (6.9%) were less frequent; Kaposi's sarcoma was notable for its absence. When the relationship between the number of oral lesions with age, risk group, medication taken, CDC staging and CD4+ count of the study group was investigated, a significantly higher number of oral lesions was associated with use of AZT, homosexuals and CDC stage IV; in contrast, a smaller number of lesions was found in those on antiparasitics and multivitamins (all P<0.05). When compared with studies from other parts of the world, the frequency of oral lesions appeared to be less common in the current study group. However, due to a lack of similar studies from the Asian region, especially in ethnic Chinese, it is not clear whether this difference could be attributed to racial, social or geographic factors.

Adult↗

Identification of matrix metalloproteinases and metalloproteinase inhibitors in bovine corpora lutea and their variation during the estrous cycle.

Corpora lutea (CL) were collected from cattle to study key physiologic events in angiogenesis. Our objective was to evaluate the activity of matrix metalloproteinases (MMP) and endogenous inhibitors. Corpora lutea were collected 2, 4, 6, 8, 10, 12, 14, and 16 d (n = 3/d) after estrus was first detected. In zymograms, a band of protein migrating at a relative molecular mass (M(r)) of 98 kDa was increased early in the cycle; a M(r) = 88 kDa band was detectable on all days. The molecular weights of these proteins are consistent with the MMP-9 family members. In all samples, a band of enzyme activity was detected at M(r) = 62 kDa, and another band of lesser density was detected at M(r) = 60 kDa. The molecular weights of these proteins are consistent with the MMP-2 family members. An immunoreactive band, detected in all samples with equal density, migrated between M(r) = 27 and 29 kDa, as did the tissue inhibitor of metalloproteinase (TIMP-1) standard. A second band, which was less dense in samples from d 2 through 6, migrated at M(r) = 19 kDa, as did the TIMP-2 standard. A third band was detected in all samples; it migrated at M(r) = 35 kDa, as did the cartilage-derived inhibitor (CDI) standard, and was less dense in d 8 and d 12 through 16 samples. In summary, MMP (gelatinases) and MMP inhibitors are present in developing luteal tissue, and the M(r) = 98 kDa enzyme, CDI, and TIMP-2 varied during the estrous cycle.

Animals↗

Four-day-old bovine corpus luteum: progesterone production and identification of matrix metalloproteinase activity in vitro.

Little is known about matrix metalloproteinases (MMPs), enzymes that are required for the structural remodeling and angiogenesis that occur in the corpus luteum (CL) during the first several days postovulation. In fact, little attention has focused on early CL function including the regulation of progesterone (P4) production. Thus, the objective of the present study was 1) to investigate the effects of insulin, LH, and dibutyryl cAMP on P4 production and cell numbers and 2) to identify MMPs in the 4-day-old CL, with use of a defined culture system. Cultures were seeded with either 1 x 10(6) or 0.5 x 10(6) cells. All cultures containing insulin had higher P4 levels and cell numbers (p < 0.05) than those without. In cultures containing insulin, basal P4 levels were high throughout the culture period. Furthermore, neither LH nor dibutyryl cAMP stimulated P4 production (p > 0.05) at a seeding density of 1 x 10(6), whereas they stimulated P4 production (p < 0.05) at a seeding density of 0.5 x 10(6) on Days 6 and 8 of culture. In conditioned medium of control cultures seeded with 0.5 x 10(6) cells, substrate gel electrophoresis (zymography) showed two intense bands that migrated at M(r) approximately 97,000 and approximately 65,000-64,000, while two weaker ones migrated at M(r) approximately 88,000 and approximately 64,000-63,000. The molecular weights of the M(r) approximately 97,000 and approximately 88,000 species were consistent with MMP-9 family members, while the molecular weights of the M(r) approximately 65,000-64,000 and approximately 64,000-63,000 species were consistent with MMP-2 family members.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Biotypes of oral Candida albicans isolates in human immunodeficiency virus-infected patients from diverse geographic locations.

Oral Candida albicans isolates from HIV-infected individuals in Hong Kong, Australia, Germany and England were characterised using a biotyping system based on enzyme profiles, carbohydrate assimilation patterns and boric acid resistance of the yeasts. A total of 44 biotypes were found amongst the 117 oral C. albicans isolates examined. The major biotype A1R accounted for 17.9% of all isolates while the second commonest biotype was A1S (11.1% of isolates). Whereas these two biotypes were isolated from all the regions studied, there were a number of other biotypes unique to individual countries. The data indicate that there are many different sub-strains of oral C. albicans in HIV-infected patients, some of which are globally prevalent. However, further work is required to ascertain the diversity of oral C. albicans biotypes, if any, in health and disease.

AIDS-Related Opportunistic Infections↗

Isolation and characterization of a complementary deoxyribonucleic acid insert encoding bovine aromatase cytochrome P450.

Aromatase, an enzyme complex comprised of aromatase cytochrome P450 (P450arom; the product of the CYP19 gene) and the flavoprotein NADPH-cytochrome P450 reductase, catalyzes the conversion of androgens to estrogens. Three cDNA inserts encoding P450arom were isolated from a bovine placental cDNA library. These inserts were sequenced and found to correspond closely to human P450arom sequence from the internal EcoRI restriction site (exon III) through the termination codon (exon X) into the 3'-untranslated region. The rapid amplification of cDNA ends technique was used to generate the rest of the cDNA 5' of the internal EcoRI site, using mRNA obtained from bovine granulosa cells as a template. This insert was sequenced, and when aligned with the other inserts, an open reading frame was found which was predicted to encode a protein of 503 amino acid residues. The deduced polypeptide shares 84% identity with human P450arom and 79%, 76%, 71%, and 57% identity with mouse, rat, chicken, and trout P450arom, respectively. A full-length open reading frame was generated using the polymerase chain reaction and mRNA obtained from bovine granulosa cells as template. After this insert was ligated into the pCMV5 expression vector, it was transfected into COS-1 monkey kidney tumor cells. We were able to demonstrate aromatase activity by assaying the incorporation of tritium into [3H] water from [1 beta-3H]androstenedione. Northern analysis revealed a single transcript of approximately 6 kilobases in poly(A)+ RNA obtained from bovine placental tissue and granulosa cells. This indicated for the first time a correspondence between the pattern of estrogen biosynthesis throughout the bovine ovarian cycle and the levels of transcripts encoding P450arom. In addition, weak hybridization was noted to transcripts of the expected size, namely 3.4 and 2.9 kilobases, in poly(A)+ RNA obtained from human placental tissue. The large size of the bovine transcript is due to a long 3'-untranslated region, because, based on the rapid amplification of cDNA ends technique, there appeared to be approximately 150 basepairs 5' of the start site of translation, and we were never able to find a polyadenylation site, even in one clone that went well past the corresponding polyadenylation site in human P450arom.

Amino Acid Sequence↗

Regulation of ovarian steroidogenesis in vitro in the viviparous shark, Squalus acanthias.

We investigated the steroid biosynthetic capabilities of ovarian granulosa and thecal elements of the viviparous dogfish, Squalus acanthias. In this report we present evidence that granulosa cells secrete quantitatively important amounts of progesterone (P), testosterone (T), and estradiol-17 beta (E), while theca has a more limited capacity to synthesize T and E. Ovarian granulosa cells were obtained from animals at each stage of gestation. After collagenase dispersion, an aliquot of 250,000 cells was incubated at 18 degrees C in basal medium, containing Eagle's salts, glutamine, penicillin, streptomycin and adjusted with 136 mM sodium chloride and 350 mM urea. After a 4 hour incubation, the content of P, T, and E in medium was determined by radioimmunoassay. P was not detectable at any time, while E was present throughout the cycle, being maximal when gestation is three quarters complete (Stage C). T gradually increased from Stage B toward late pregnancy. In Stage C granulosa cells, E production increased in the presence of graded doses of T substrate. Also, a homologous pituitary extract (1/25 equivalents) and the calcium ionophore A23187 stimulated production of all 3 steroids. Using radioisotopes, granulosa cells showed a wide range of synthetic capacities. In Stage C thecal tissue, E production also increased in the presence of graded doses of T substrate, while pituitary extract only increased T. When granulosa and theca were recombined, in the presence of pituitary extract, P levels decreased with a corresponding increase in T, when compared to granulosa alone. These data suggest a possible interaction between granulosa and theca for steroid biosynthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Oxytocin-specific RNA, oxytocin and progesterone concentrations in corpora lutea of heifers treated with oxytocin.

Sixteen virgin Holstein heifers were used to study the role of oxytocin in luteal function. Oxytocin (100 i.u.) was administered on Days 2-5 (Exp. 1) or Days 2-6 (Exp. 2), and the corpora lutea were collected and weighed on Day 6 or Day 8, respectively. Plasma progesterone values and tissue concentrations of progesterone and oxytocin were determined by radioimmunoassay. In addition, the tissue level of the oxytocin-specific transcript was measured by dot-blot hybridization. Expression of the oxytocin-specific transcript was not affected by oxytocin treatment in either experiment. In contrast, plasma progesterone values and tissue progesterone and oxytocin concentrations were all decreased after 5 days of oxytocin treatment (Exp. 2), but not after 4 days (Exp. 1). Since oxytocin concentrations were reduced while the level of its transcript remained unaffected, oxytocin injections might be influencing either peptide processing or release.

Animals↗

In vivo steroidogenic effects of homologous pituitary ventral lobe extract in the spiny dogfish, Squalus acanthias.

Pregnant Squalus acanthias were injected intravenously with aqueous extract equivalent to three pituitary ventral lobes (VL). Blood samples were withdrawn by caudal sinus puncture at intervals over a period of 72 hr and plasma progesterone, testosterone, and estradiol-17 beta measured by radioimmunoassay. Early pregnancy animals (Stage A) failed to respond, but mid-pregnancy animals (Stage C) responded to VL with significant increases in plasma steroids. Plasma progesterone and testosterone were increased sevenfold and plasma estradiol-17 beta fourfold by 10-24 hr postinjection. These studies demonstrate pregnancy stage-dependent differences in gonadal sensitivity to a pituitary ventral lobe gonadotropic moiety.

Animals↗

Morphological and endocrine correlates of the reproductive cycle of the aplacental viviparous dogfish, Squalus acanthias.

Pregnant female spiny dogfish were captured off the coasts of Maine and Massachusetts and morphometric and gravimetric analyses of the liver, gonad, and reproductive tract correlated with the reproductive cycle. Plasma samples were taken and circulating steroids (progesterone, testosterone, and estradiol) were measured by radioimmunoassay. Clear-cut temporal variations in plasma steroid levels were observed, which correlated with the ovarian cycle. Testosterone and estradiol-17 beta levels were low (100-200 pg/ml plasma) in early pregnancy when ovarian follicles were small but increased to about 10 ng/ml as follicles grew and gestation progressed. By contrast, plasma progesterone was high (2-6 ng/ml) in early to mid-pregnancy when corpora lutea were active and declined toward term (less than 1.0 ng/ml).

Animals↗