RESUMEN
INTRODUCTION: Methylenetetrahydrofolate reductase (MTHFR) C677T is involved in folate and homocysteine metabolism. Disruption in the activity of this enzyme will alter their levels in the body. METHODOLOGY: This study assessed MTHFR C677T polymorphism and its relationship with serum homocysteine and B-vitamins levels in a sample of Chinese and Malays subjects in UPM, Serdang. One hundred subjects were randomly selected from among the university population. Folate, vitamin B12, B6, and homocysteine levels were determined using MBA, ECLIA, and HPLC, respectively. PCR coupled with HinfI digestion was used for detection of MTHFR C677T polymorphism. RESULTS: The frequency of T allele was higher in the Chinese subjects (0.40) compared to the Malay (0.14). Folate, vitamin B12 and B6 levels were highest in the wild genotype in both ethnic groups. Subjects with heterozygous and homozygous genotype showed the highest homocysteine levels. The serum folate and homocysteine were mainly affected by homozygous genotype. CONCLUSION: MTHFR C677T polymorphism plays an important role in influencing the folate and homocysteine metabolism.
Asunto(s)
Enfermedad de la Arteria Coronaria/genética , Homocisteína/sangre , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Polimorfismo Genético , Vitamina B 12/sangre , Adulto , Pueblo Asiatico/genética , Enfermedad de la Arteria Coronaria/sangre , Enfermedad de la Arteria Coronaria/epidemiología , Estudios Transversales , ADN/análisis , Cartilla de ADN , Femenino , Humanos , Malasia/epidemiología , Masculino , Reacción en Cadena de la Polimerasa , Estudiantes , Adulto JovenRESUMEN
Introduction: Methylenetetrahydrofolate reductase (MTHFR) C677T is involved in folate and homocysteine metabolism. Disruption in the activity of this enzyme will alter their levels in the body. Methodology: This study assessed MTHFR C677T polymorphism and its relationship with serum homocysteine and Bvitamins levels in a sample of Chinese and Malays subjects in UPM, Serdang. One hundred subjects were randomly selected from among the university population. Folate, vitamin B12, B6, and homocysteine levels were determined using MBA, ECLIA, and HPLC, respectively. PCR coupled with HinfI digestion was used for detection of MTHFR C677T polymorphism. Results: The frequency of T allele was higher in the Chinese subjects (0.40) compared to the Malay (0.14). Folate, vitamin B12 and B6 levels were highest in the wild genotype in both ethnic groups. Subjects with heterozygous and homozygous genotype showed the highest homocysteine levels. The serum folate and homocysteine were mainly affected by homozygous genotype. Conclusion: MTHFR C677T polymorphism plays an important role in influencing the folate and homocysteine metabolism.
RESUMEN
Persistent high-risk human papillomavirus (HPV) infection is known to play an important role in the genesis of cervical cancer. Since new screening and prevention strategies, namely improved HPV testing and HPV vaccination have been aggressively promoted recently, it is crucial to investigate the HPV distribution in Malaysia in order to maximize their cost-effectiveness. This study was therefore conducted to assess the HPV type distribution in the most populous region, the state of Selangor. A total of 200 cervical swab samples were collected in two health-screening campaigns, and also from women attending obstetrics and gynecology clinics in several hospitals in Selangor. DNA extraction was performed and HPV DNA was detected via nested PCR using MY09/MY11 as outer primers and GP5+/GP6+ as inner primers which target the L1 gene of the viral genome. The purified PCR products were subjected to automated DNA sequencing to determine the HPV genotype. Out of 180 ß-globin positive samples, 84 (46.7%) were positive for HPV DNA. The most common HPV type found was high-risk oncogenic type 16 (40%), followed by HPV type 18 (3.3%), HPV 33 (1.7%), HPV 31 (0.6%), and low-risk HPV 87 (0.6%). Our study confirmed that nested PCR method is highly sensitive in detecting HPV DNA even in low risk patients. Since a relatively high prevalence rate of HPV infection was found in this population, prompt healthcare policy changes to bring about implementation of early HPV vaccination program is desirable to prevent a high incidence of cervical cancer.
Asunto(s)
ADN Viral/genética , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Displasia del Cuello del Útero/epidemiología , Neoplasias del Cuello Uterino/epidemiología , Adulto , Anciano , Secuencia de Bases , Femenino , Humanos , Incidencia , Malasia/epidemiología , Persona de Mediana Edad , Datos de Secuencia Molecular , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/virología , Proyectos Piloto , Reacción en Cadena de la Polimerasa , Prevalencia , Pronóstico , Factores de Riesgo , Homología de Secuencia de Ácido Nucleico , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/virología , Frotis Vaginal , Adulto Joven , Displasia del Cuello del Útero/genética , Displasia del Cuello del Útero/virologíaRESUMEN
The cholera enterotoxin (CT) has been considered a major virulence factor of Vibrio cholerae. The accessory cholera enterotoxin (ace) gene is the third gene of V. cholerae virulence cassette. The gene coding for the Ace toxin was amplified from V. cholerae isolates producing a single band of 314 bp. The presence of ace gene was confirmed by hybridization as well as by sequencing. The gene was successfully expressed in Escherichia coli (LMG194) using expression, pBAD/Thio-TOPO vector. Optimal conditions for expression included choice of host strain, temperature used for culturing, and concentration of antibiotic and arabinose inducer. The Ace protein was obtained from the cell supernatant as a fusion protein with a molecular mass 34 kDa which was detected using an anti V5-HRP epitope tagged antibody.