Comparison of human papillomavirus genotyping using commercial assays based on PCR and reverse hybridization methods

Different tests for human papillomavirus (HPV) screening are commercially available, detecting high‐risk oncogenic HPV types with a pool of genotype‐specific probes. However, it is necessary to establish reliable methods for the identification of individual genotypes. The purpose of this study was to compare three different commercial methods for HPV genotyping: INNO‐LiPA HPV Genotyping v2 (LiPA), Linear Arrays HPV Genotyping Test (LA) and Clinical Arrays Human Papillomavirus (CA). A total of 83 HPV DNA‐positive samples by hybrid capture method were genotyped (82, 78 and 81 by LiPA, LA and CA, respectively). Comparison analysis was limited to the HPV genotypes common to the three assays. There were concordant results (absolute agreement between assays) in 31 samples (39.7%) and compatible results (correspondence for some but not all genotypes) were found in 44 samples (56.4%). Only three samples (3.8%) were considered as discordant (did not show any similarity between the tests). Analyzing kappa values we have a very good agreement (>0.8) for HPV16 and HPV31 and good agreement (0.6–0.8) for HPV types 6, 18, 53 and 66 when all methods are compared. We conclude that all genotyping methods tested are highly comparable and suitable for clinical and epidemiological studies.     

KNEE PAIN AMONGST THE POOR AND AFFLUENT IN PAKISTAN

The frequency of joint symptoms was determined amongst 2022affluent and 2210 poor adults in Karachi, Pakistan. Joint painwas significantly (P = 0.025) more common amongst the affluent(6.6%) compared with the poor (5%) and this was due to a significantlygreater frequency of knee pain in the richer community (3% vs1.8%; P =0.008). The prevalence increased with age and was morecommon in females. Almost half were associated with varus deformity,suggesting the presence of associated OA in a high proportion.The overall frequency of knee pain seemed no greater than inseries reported from the West. Compared with age- and sex-matchedcontrols, body weight was significantly greater amongst thosewith knee pain, both amongst the affluent (P = 0.005) and thepoor (P = 0.02). Control subjects were heavier in the affluentpopulation, suggesting that the greater frequency of knee symptomsin this community was due to their relative obesity. Knee bendingat prayer was most common amongst the affluent controls andmay indicate that religious observance also contributed to theproblem in the richer population. Squatting was a characteristicof the poor who had less knee pain than the affluent. Knee flexingcould not therefore be confidently implicated. No relationshipcould be demonstrated between knee pain and joint laxity. KEY WORDS: Knee pain, Osteoarthritis, Pakistan, Obesity, Knee flexing, Joint laxity     

FAILURE OF HIGH-DOSE SUSTAINED RELEASE LUTEINIZING HORMONE RELEASING HORMONE AGONIST (BUSERELIN) PLUS ORAL TESTOSTERONE TO SUPPRESS MALE FERTILITY

Previously we have demonstrated that sperm counts of normal young men decreased during constant subcutaneous infusion of the LHRH agonist buserelin (118 or 230 micrograms/d). In order to test whether azoospermia can be achieved with higher doses, seven young men received 450 micrograms buserelin subcutaneously daily for 12 weeks via extracorporeal osmotic minipumps. To avoid symptoms of androgen deficiency, oral supplementation with 80 mg/d testosterone undecanoate (TU) was initiated in week 5 and was increased to 120 mg/d by week 8. Follow-up after treatment lasted for another 12 weeks. In order to evaluate possible psychotropic effects of treatment-related endocrine changes, continuous psychometric testing was performed focusing on personality, emotions and sexuality. After an initial rise, both serum LH and FSH returned to normal. FSH was below normal during the 3rd-5th week following treatment. LHRH stimulation tests performed at the end of treatment showed pituitary desensitization. Serum T (always measured between 0800 and 1300 h at least 12 h after last TU) tended to decrease by week 7 and remained slightly depressed until the end of treatment while libido, potency and emotional well-being remained unchanged. While testicular volumes showed a reduction from week 7 of treatment to week 10 post-treatment, sperm counts decreased only insignificantly from 65 +/- 10 to 44 +/- 14 million per ml in week 12 post-treatment. Severe oligo- or azoospermia was not observed in any of the seven men. It is concluded that full androgen substitution by TU can drastically delay if not abolish the antifertility effect of LHRH-induced pituitary desensitization.     

Effects of retinol on glycoprotein synthesis by Sertoli cells in culture: dolichyl phosphomannose synthase activation

Sertoli cells were isolated from Wistar rats aged 19 days and cultured for 48 h. The addition of retinol (10 μM) to the culture medium significantly stimulated the incorporation of [2–³H]mannose into lipid-linked oligosaccharide and into cellular and secreted glycoproteins. Incorporation of [U- 14C] leucine into proteins and of [5, 6–³ H] uridine into RNA was unaffected by retinol treatment. Incubation of microsomal fractions of retinol-treated cells showed an increase in mannose incorporation into dolichyl phosphomannose, into dolichyl pyrophosphoryl oligosaccharide and into proteins. Chromatographic analysis of the fraction soluble in chloroform/methanol (2: 1 v/v) did not show the presence of retinyl phosphomannose either in control or in retinol-treated cells. When the formation of dolichyl phosphomannose was studied in microsomes isolated from control cells and from cells treated with 10 μM retinol for 48 h in the presence of exogenous dolichyl phosphate, the results showed that the retinol effect was due to stimulation of dolichyl phosphomannose synthase.