CYP3A5*3 Genetic Polymorphism and Tacrolimus Concentration in Myanmar Renal Transplant Patients.

BACKGROUND: Genetic polymorphism is an important factor that influences tacrolimus concentrations and has the potential to predict the optimal dosage of tacrolimus in personalized medicine. Tacrolimus, a drug of narrow therapeutic index, is used in renal transplant recipients as an immunosuppressant agent. It is a substrate of cytochrome P450 3A (CYP3A) and has highly variable pharmacokinetic parameters. OBJECTIVE: The aim of this study was to identify the proportion of CYP3A5 gene polymorphism in Myanmar kidney transplant recipients and to determine the impact of CYP3A5 gene polymorphisms on tacrolimus level in CYP3A5 expressors and nonexpressors. METHODS: This study included 41 adult Myanmar post-renal transplant patients. Tacrolimus trough blood levels were determined and CYP3A5 genotype analysis was conducted by using polymerase chain reaction amplification of target followed by detection by restriction fragment length polymorphism analysis. RESULTS: The CYP3A5 nonexpressors and expressors were detected in 25 (60.97%) and 16 (39.02%) of the 41 renal transplant recipients, respectively. The tacrolimus concentration/dose ratio in the CYP3A5 expressor group was lower than in the CYP3A5 nonexpressor group (1.49 ± 0.69 vs 3.49 ± 3.08 [P = .003] at 1 month; and 1.54 ± 0.9 vs 7.88 ± 8.25 [P = .0001] at 3 months). CONCLUSIONS: The findings of the present study revealed that more than one half of the study population were carrying the mutant allele CYP3A5*3(A6986G). CYP3A5 genetic polymorphism is one of the important factors in determining daily requirements for tacrolimus and in adjusting tacrolimus trough concentrations.

HIV type 1 subtypes in Malaysia, determined with serologic assays: 1992-1996.

We investigated the molecular epidemiology of HIV-1 subtypes in Malaysia among injecting drug users (IDUs) and sexual transmission risk groups, using serologic and genetic techniques. Frozen sera collected at a general hospital, a blood bank, several drug treatment centers, and an STD clinic in Kuala Lumpur, between 1992 and 1996, were investigated retrospectively. V3 peptide serotyping and monomeric gp120 capture serotyping were used to study 89 known HIV-1-infected subjects. The methods differentiate subtypes B, E, and C. V3 peptide and gp120 capture results were comparable. No subtype C-specific reactive sera were found; one specimen was dually reactive for subtypes C and B, using the V3 peptide ELISA; and four were durally reactive for subtypes E and C using this assay. Genotypic analysis of HIV-1 gag RNA in serum was done on a subset of subjects and confirmed serologic findings. HIV-1 subtypes differed significantly by risk category: of 53 IDUs, 29 (55%) were infected with subtype B and 19 (36%) were infected with subtype E, 3 (6%) were dually reactive, and 2 (4%) were not typable. Of 36 persons with heterosexual risks, 29 (81%) were infected with subtype E, 5 (14%) were infected with subtype B, and 2 (5%) were not typable. Persons with IDU risks were significantly more likely to be infected with subtype B than were those with sexual risks (OR 5.89; 95% CI, 1.94-18.54; p < 0.001). Subtypes B and E of HIV-1 appear to predominate in Malaysia; subtype B was more prevalent among IDUs; subtype E was more prevalent among all other groups. These results may have important HIV-1 vaccine implications.

Reinfection of people with Ascaris lumbricoides following single, 6-month and 12-month interval mass chemotherapy in Okpo village, rural Burma.

A longitudinal study on reinfection with Ascaris lumbricoides was continued on a random sample of 50% of the infected population following a horizontal cross-sectional study in Okpo village, near Rangoon. The study sample was again randomly divided into two subsamples, the six-month interval worming group and the 12-month interval worming group. Microscopic examination of stool for Ascaris eggs on the 7th day and 30th day, combined with counting eggs and worming with levamisole, were carried out at two successive six-month intervals on the first subsample. Stools were examined and eggs counted on the 7th day and then monthly up to 12 months, followed by worming, in the second subsample. The worms expelled in the first 72 hours after treatment were counted. The findings over a 12-month follow-up period after one treatment included: mean monthly incidence of 20%, higher and more rapid return to previous prevalence and intensity of infection in children or 'wormy' persons than in adults or 'non-wormy' persons. Six-monthly chemotherapy definitely reduced intensity of infection in the children and adults whereas 12-monthly treatment lowered intensity in adults only. Predisposition to acquisition of high or low intensity of infection was also observed. Other findings and the implications of this study for strategies for control of ascariasis are discussed.

Evaluation of a rapid test for the detection of antibodies to human immunodeficiency virus type 1 and 2.

Hema-Strip HIV-1/2 is a one-step rapid test for the detection of anti-HIV-1/2 antibodies in whole blood. The test requires no expensive equipment and the results are available within 10-15 min. Using 72 known HIV-1 positive samples and 780 high-risk prisoners, the sensitivity and specificity of Hema-Strip HIV-1/2 was found to be comparable to microparticle enzyme immunoassay (MEIA). The data also indicated that Hema-Strip HIV-1/2 is an effective alternate testing system to conventional ELISA where the use of ELISA is not suitable and the result of the HIV testing is needed urgently.

Footballer's hip a report of six cases.

We have reviewed a consecutive series of six professional footballers who presented with intractable hip pain which was attributed at arthroscopy, to an anterior acetabular labral tear with adjacent chondral damage. The location and type of labral tear were identical for each patient. There was no evidence of acetabular dysplasia. A variable area of chondral damage was associated with the labral tear. All unstable tissue was resected. Five returned to professional football at the highest level. Acetabular labral pathology should be included in the differential diagnosis of footballers with hip or groin pain. Arthroscopy of the hip is an appropriate method of diagnosis and treatment and minimises the length of rehabilitation required.

Composite cure and pulp-cell cytotoxicity associated with LED curing lights.

This study compared the cure and pulp-cell cytotoxicity of composites polymerized with light-emitting diode (LED) and halogen-based light curing units. A mini-filled resin composite (Tetric Ceram, Vivadent), two LED (E-light [EL], GC and Freelight [FL], 3M-ESPE), a conventional halogen (Max [MX], Dentsply) and a high-intensity halogen light (Astralis 10 [AS], Vivadent) were evaluated. Cure associated with the different lights was determined by measuring the top and bottom surface hardness (KHN; n = 5) of 2-mm thick specimens using a digital microhardness tester (load = 500 gf; dwell time = 15 seconds). Pulp-cell cytotoxicity was assessed using a direct contact method involving incisor tooth slices dissected from 28-day old Wistar rats maintained in Dulbecco's Modified Eagle's Medium (DMEM) and 1% agarose. The bottom surfaces of the cured composite specimens (7-mm diameter and 2-mm deep) were placed in contact with the openings of each tooth slice. After incubation in 5% CO2 atmosphere at 37 degrees C for 48 hours, the tooth slices were fixed, demineralized and processed for histological examination. Pulp fibroblasts and odontoblasts were counted histomorphometrically at 400x magnification within a 1500 microm2 area using a computerized micro-imaging system. Eighteen readings were obtained for each curing light. Data was subjected to ANOVA/Scheffe's test and Pearson's correlation at significance level 0.05 and 0.01, respectively. At the top surfaces, the cure with AS was significantly greater than the other curing lights, with MX and FL being significantly greater than EL. At the bottom surfaces, MX, AS and FL had significantly better cure than EL. Specimens cured with MX were less cytotoxic than those polymerized with other curing lights. Specimens cured with AS and EL were significantly less cytotoxic than FL. Composite cure and cytotoxicity associated with LED lights is device dependent. Composite cure was not correlated to pulp-cell cytotoxicity. The response of pulpal fibroblasts to unreacted/leached components of composites differs somewhat from odontoblasts.

Concurrence of hepatitis B surface antigen and antibody in acute and chronic hepatitis B cases.

A total of 250 hepatitis B surface antigen positive sera were screened for antibody to hepatitis B surface antigen. It was found that seven (3%) sera showed concurrently circulating surface antigen and surface antibody to hepatitis B virus. The level of antibody to surface antigen was not affected by HBeAg and most of the cases were found in chronic hepatitis B carriers.

Hepatitis B surface antigen subtypes in hepatitis B seropositive subjects in University Hospital, Kuala Lumpur.

Hepatitis B surface antigen can be serologically defined as ayw1, ayw2, ayw3, ayw4, ayr, adw2, adw4 and adrq+ or adrq-. A study of common HBsAg subtypes in 44 HBsAg reactive sera in University Hospital was conducted using a solid-phase sandwich EIA. Eleven samples were found not typable and among the 33 typable HBsAg reactive sera, 3 HBsAg subtypes: adw, adr and ayw were identified. Subtype adw was found in 66.7% (22/33) of the typable HBsAg reactive sera; 24.2% (8/33) was of subtype adr and 6.0% (2/33) of subtype ayw. One sample was found to be reactive to both adw and adr. HBsAg subtype adw was found more commonly in Chinese but among the Malays, HBsAg subtype adr appeared to predominate. However, the small sample size precludes firm conclusions on the predominant subtype among the Malays.

Evaluation of three commercial rapid tests for detecting antibodies to human immunodeficiency virus.

Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest are simple/rapid tests for the detection of antibodies to HIV-1 and HIV-2 in human whole blood, serum and plasma samples. The assay is one step and the result is read visually within 15 minutes. Using 92 known HIV-1 reactive sera and 108 known HIV-1 negative sera, the 3 HIV tests correctly identified all the known HIV-1 reactive and negative samples. The results indicated that Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest HIV are as sensitive and specific (100% concordance) as Microparticle Enzyme Immunoassay. The data indicated that these 3 HIV tests are effective testing systems for diagnosis of HIV infection in a situation when the conventional Enzyme Immunoassay is not suitable.

A seroprevalence study of viral hepatitis E infection in human immunodeficiency virus type 1 infected subjects in Malaysia.

Hepatitis E virus (HEV) is a RNA virus transmitted enterically. A study of anti-HEV antibodies in 145 human immunodeficiency virus type 1 (HIV-1) infected subjects found that 14.4% of them were reactive to anti-HEV antibodies. Anti-HEV IgG and anti-HEV IgM was detected in 10.3% and 4.1% of the subjects respectively. Prevalence of anti-HEV (either IgG or IgM) was similar across all adult ages (p = 0.154), between the three ethnic groups (p = 0.378), and across risk groups (p = 0.120). The results showed that HEV infection in subjects recruited in this study was most likely transmitted via faecal-route.

The prevalence of anti-HCV antibody in risk groups and blood donors.

Anti-HCV antibody was detected in 1.9% of the blood donors in University Hospital. Among the risk groups, 33.3% of the patients with post-transfusion hepatitis were tested positive for anti-HCV antibody. The anti-HCV antibody was detected in 30% of the IDU. Haemodialysis patients, patients with acute and chronic hepatitis and patients with liver cirrhosis appeared to have increased risk of Hepatitis C virus infection. The results indicate that the frequency of HCV infection increases with the exposure to blood or blood products.

Effects of berberine, a plant alkaloid, on the growth of anaerobic protozoa in axenic culture.

Extracts obtained by organic solvents from the root of Coptis teeta in Myanmar (Burma), were tested for growth inhibitory activity against Giardia lamblia, Trichomonas vaginalis and Entamoeba histolytica in axenic culture. All extracts had anti-protozoal activity with the methanol extract, in particular, being effective against all 3 parasites. By chromatographic analysis, the extracts were shown to contain berberine compounds and other alkaloids. Comparing the inhibitory effects of the methanol extract with berberine sulfate, a berberine salt previously shown to possess inhibitory activity, it was observed that the crude extract was more effective than the salt. This suggests that the greater inhibitory activity of the crude extract may be due to the cumulative contributions of its contained berberines and alkaloids. Based on these findings, it appears that the berberine compounds may be useful as chemotherapeutic agents against the 3 parasites tested.

Impact of the Expanded Program of Immunization against hepatitis B infection in school children in Malaysia.

The implementation of the Expanded Program of Immunization (EPI) in 1989 has dramatic impact on hepatitis B virus (HBV) infection in school children in Malaysia. A cross-sectional seroprevalence study of HBV infection in 190,077 school children aged 7-12 years from 1997 to 2003 showed a steady decline of HBV surface antigen (HBsAg) prevalence rate from 2.5% for children born in 1985 to 0.4% among school children born in 1996. The overall prevalence of HBsAg was 0.6%, 0.7% in males and 0.6% in females. Over 92.7% of school children had been vaccinated with HBV vaccine, in which 93.7% were vaccinated under the EPI and 6.3% on voluntary basis. The school children vaccinated under EPI had a 0.4% HBsAg carrier rate, which was significantly lower than school children vaccinated on a voluntary basis (HBsAg carrier rate 1.3%) and non-vaccinated school children (HBsAg carrier rate 2.7%), suggesting that HBV vaccination of infants was the most effective measure in preventing vertical transmission of HBV in the hyperendemic region.

Results of hybrid total hip replacement and resurfacing - is there a difference?

Hip resurfacing has in recent years become increasingly popular as an arthroplasty technique, particularly amongst younger patients with more active lifestyles. The procedure has been heralded as a more conservative option, aimed at those wishing to continue strenuous work and recreational activities. Little evidence has appeared in the literature however, either with the results of resurfacing or comparing resurfacing to conventional total hip arthroplasty. We compare the outcomes of a consecutive series of hybrid hip resurfacings against a similar series of conventional hybrid hip arthroplasties. In our series there was no difference in operation time, blood loss, or modified Harris Hip score at 12 months postoperatively. There was a small decrease in length of stay, which was not clinically significant. We conclude that whilst it has been previously shown that there is an overall preservation of bone stock following resurfacing arthroplasty, there is no evidence to back additional claimed benefits. (Hip International 2004; 14: 169-73).

Systemic Candida infection in University hospital 1997-1999: the distribution of Candida biotypes and antifungal susceptibility patterns.

A total of 102 Candida species were isolated from blood cultures from January 1997 to October 1999. Using assimilation of carbohydrate test, 52 (51.0%) of the Candida sp. were identified as C. parapsilosis, 25.5% (26) were C. tropicalis. C. albicans made up 11.8% (12), 6.9% (7) were C. rugosa, 3.8% (4) C. glabrata and 1% (1) C. guilliermondii. No C. dubliniensis was found in the study. In vitro antifungal susceptibility tests showed that all Candida species were sensitive to nystatin, amphotericin B and ketoconazole. Although all isolates remained sensitive to fluconazole, intermediate susceptibility was found in 3 C. rugosa isolates. Antifungal agents with high frequency of resistance were econazole, clotrimazole, miconazole and 5-fluorocytosine. Candida species found to have resistance to these antifungal agents were non-C. albicans.

Onychomycosis in Malaysia.

The common etiological agents of onychomycosis are dermatophytes, molds and yeasts. A mycological nail investigation of onychomycosis using direct microscopy and culture was conducted by the Mycology Unit, Department of Medical Microbiology, University of Malaya from March 1996 to November 1998. The study involved 878 nail clippings or subungal scrapings from subjects with onychomycosis. On direct microscopy examination, 50% of the specimens were negative for fungal elements. On culture, 373 specimens had no growth; bacteria were isolated from 15 nail specimens. Among the 490 specimens with positive fungal cultures, 177 (36.1%) were dermatophytes, 173 (35.5%) were molds and 130 (26.5%) were Candida. There were 2% (10/490) mixed infections of molds, yeasts and dermatophytes. Trichophyton rubrum (115/177) and Trichophyton mentagrophytes (59/177) were the main dermatophytes isolated. The molds isolated were predominantly Aspergillus niger (61/173), Aspergillus nidulans (30/173), Hendersonula toruloidea (26/173) and Fusarium species (16/173). 96.9% of the Candida species identified were Candida albicans.

Outbreak of Vibrio cholerae 01 in Hong Kong related to contaminated fish tank water.

An outbreak of 12 cholera cases, caused by Vibrio cholerae eltor inaba, occurred in Hong Kong during a three week period in June-July 1994. Only adults of both sexes were affected. Epidemiological investigations showed linkage in all cases with consumption of seafood, including shellfish, mantis shrimps and crabs. Microbiological findings demonstrated that contaminated seawater in fish tanks used for keeping alive these seafoods is the most likely vehicle of transmission. Aggressive control measures, promptly instituted, included prohibition of use of contaminated typhoon shelter water in fish tanks, use of seawater with E. coli counts below 610 organisms/100 ml, and the banning of unlicensed food sampans in typhoon shelters. These measures, coupled with public announcements and an active health education campaign on food safety and personal hygiene, abruptly terminated the outbreak. Places which practise the use of seawater, from probable contaminated sources, to keep alive their seafood for human consumption should be alerted to the possibility of transmission of Vibrio cholerae through this route.

Changes in levels of anti-dengue virus IgG subclasses in patients with disease of varying severity.

Extensive complement activation precedes onset of shock in dengue patients and complement "split products" C3a and C5a could be responsible, directly or indirectly, for the increased vascular permeability and disseminated intravascular coagulation which characterises dengue haemorrhagic fever (DHF) dengue shock syndrome (DSS). As IgG subclasses vary in their capacity to activate the classical complement pathway after combining with antigen, we have used an indirect enzyme linked immunosorbent assay (ELISA) to assess levels of IgG1-4 against each dengue serotype in acute and convalescent sera from patients with disease of varying severity. Acute phase sera from patients with dengue haemorrhagic fever (DHF) or dengue shock syndrome (DSS) contained higher levels of anti-dengue antibodies of the IgG1, complement fixing, subclass than similar sera from dengue fever (DF) patients. Conversely, acute phase sera from DHF and DSS patients contained lower levels of anti-dengue antibodies of the poor complement activating IgG2 subclass than acute phase sera from DF patients. No significant differences were detected between the levels of anti-dengue IgG3 and IgG4 antibody in acute phase sera from DF, DHF, and DSS patients. With the exception of levels of anti-dengue IgG2 antibody from DHF patients which were lower than those from DF and DSS patients, levels of anti-dengue IgG1, IgG2, IgG3, and IgG4 were similar in convalescent sera from all patients. These results provide a possible explanation for the activation of the serum complement system which precedes onset of shock in severe dengue infections.

Candida biotypes isolated from clinical specimens in Malaysia.

The distribution of Candida species was examined using 1114 yeasts isolated from various clinical specimens. The isolates were identified by germ tube test, hyphal/pseudohyphae and chlamydoconidia production and carbohydrate assimilation test using ten carbohydrates (glucose, sucrose, trehalose, cellobiose, arabinose, galactose, mannitol, raffinose, lactose and maltose). Among the 1114 isolates studied, 9 species of Candida were identified and the relative frequency of isolation was C. albicans (44.2%), C. parapsilosis (26.0%), C. tropicalis (17.7%), C. glabrata (9.6%), C. krusei (1.2%), C. rugosa (0.6%), C. guilliermondii (0.2%), C. lusitaniae (0.08%) and C. kefyr (0.08%). Non-C. albicans was the most common Candida species isolated from blood, respiratory system, urine and skin. The isolate from vaginal swabs was predominantly C. albicans. 82.2% of C. glabrata and 64.2% of C. krusei isolated in this study were from vaginal swabs.