Sabariah Abdul Rahman

Assessing estrogenic activity of Nigella sativa in ovariectomized rats using vaginal cornification assay

The aim of this study was to determine the estrogenic activity of Nigella sativa (NS) by vaginal cornification assay using an ovariectomized rat model. Forty ovariectomized Sprague Dawley rats, weighting 250 to 350 g were used in the study. N. sativa powders were administered to ovariectomized rats for 21 consecutive days at dosages of 300, 600 and 1200 mg/kg respectively, and were compared with each of daily treatment with 0.2 mg/kg conjugated Equine estrogen as positive control or distilled water as negative control. Vaginal smears were taken to observe the estrogenic effect on vaginal epithelium of rats. The vaginal smear showed an atrophic pattern at baseline. The occurrence of vaginal cornification after N. sativa supplementation indicated estrogenic activity of N. sativa, but this effect was not as much as CEE. The most influence of N. sativa in vaginal cornification was observed in low dose NS that this result was in agreement with serum Estradiol level of this group. The cornified cells percentage significantly differed from control group (P<0.05). These data suggest that N. sativa possesses estrogenic function in the ovariectomized rat model which can be helpful in managing menopausal symptoms as an alternative for Hormone Replacement Therapy.

p16 Gene Expression in Basal Cell Carcinoma

BACKGROUND Basal cell carcinoma (BCC) develops predominantly in sun-exposed skin in fair-skinned individuals prone to sunburn. BCC typically occurs in adults. High exposure to ultraviolet (UV) radiation increases rate of developing BCC, a slowly growing tumor that occurs in hair-growing squamous epithelium and rarely metastasizes. In genetic studies, BCC patients have cell-cycle abnormalities of different parts of the signaling pathway. Retinoblastoma regulatory pathway is important in cell cycle arrest. In this pathway, p16INK4a, an inhibitor of Rb pathway, binds to CDK4 and CDK6 competitively with cyclin D1 to prevent phosphorylation of tumor suppressor pRB gene. Alteration of this pathway contributes to development of human cancers and also is effective in skin cancers. In this study, we analyzed mRNA expression using in situ RT-PCR and the role of immunohistochemical expression of p16INK4a in BCC. METHODS Expression of p16 in ten samples of Iranian paraffin-embedded skin BCC were studied using in situ RT-PCR and immunohistochemistry on p16INK4a gene. RESULTS Nuclear and cytoplasmic staining intensity of samples within tumor cells and normal skin tissue illustrates different mRNA and protein expression of p16 gene. mRNA of p16 gene and the expressed protein induce cell cycle proliferation and involve both tumor tissue as well as normal skin tissue. However, in this study it was found that there is significant protein and mRNA expression in BCC cells when compared to normal skin tissue (p<0.05). CONCLUSIONS p16 gene is involved in the pathogenesis of human skin BCC in view of increased p16 mRNA and expressed protein within tumor cells.

Gene transfer into the lung by nanoparticle dextran-spermine/plasmid DNA complexes.

A novel cationic polymer, dextran-spermine (D-SPM), has been found to mediate gene expression in a wide variety of cell lines and in vivo through systemic delivery. Here, we extended the observations by determining the optimal conditions for gene expression of D-SPM/plasmid DNA (D-SPM/pDNA) in cell lines and in the lungs of BALB/c mice via instillation delivery. In vitro studies showed that D-SPM could partially protect pDNA from degradation by nuclease and exhibited optimal gene transfer efficiency at D-SPM to pDNA weight-mixing ratio of 12. In the lungs of mice, the levels of gene expression generated by D-SPM/pDNA are highly dependent on the weight-mixing ratio of D-SPM to pDNA, amount of pDNA in the complex, and the assay time postdelivery. Readministration of the complex at day 1 following the first dosing showed no significant effect on the retention and duration of gene expression. The study also showed that there was a clear trend of increasing size of the complexes as the amount of pDNA was increased, where the sizes of the D-SPM/pDNA complexes were within the nanometer range.

Multidrug resistance protein 2 genetic polymorphism and colorectal cancer recurrence in patients receiving adjuvant FOLFOX-4 chemotherapy

Multidrug resistance protein 2 (MRP2), encoded by the ATP-binding cassette C2 (ABCC2) gene, is an efflux pump located on the apical membrane of many polarized cells, which transports conjugate compounds by an ATP-dependent mechanism. The correlation of G1249A ABCC2 polymorphism with the development of colorectal cancer (CRC) and poor prognosis was evaluated in patients who were treated with fluorouracil/-leucovorin (FL) plus oxaliplatin (FOLFOX-4). A total of 50 paraffin‑embedded tissue samples collected from CRC patients were analyzed to identify the polymorphism. Patients were in stage II/III and received postoperative FOLFOX-4 chemotherapy. As a control group, an equal number of unrelated healthy subjects were enrolled in the study. The polymorphism was genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, and results were compared with clinicopathological markers, early relapse and survival rates. During the 12 months of follow-up, local and distant recurrences were observed in 15 (30%) patients. No significant difference in the distribution of wild-type and polymorphic genotypes was observed between the patient and control groups and between the patients who experienced recurrence within 1 year and those who did not (all P>0.05). In conclusion, the G1249A polymorphism is not associated with CRC risk and early recurrence. However, significant correlation was observed between G1249A polymorphism and the overall survival and disease-free survival of the patients.

Association Between Telomere Shortening and Ageing During Occupational Exposure

Association Between Telomere Shortening and Ageing During Occupational Exposure Telomere length is considered as a biomarker of ageing, resulting in shortening during the process. The present investigation was an attempt to determine the relative telomere length in mechanical workshop workers. Telomere length shortening in cells during occupational exposure causes accelerated ageing. Genomic DNA was isolated from buccal epithelial cells collected from 240 individuals, comprising two groups of 120 exposed workers and 120 unexposed controls. Telomere length was measured by using real time PCR. Both telomere (T) and single copy gene (S) specific primers were used to compute the relative T/S ratio and expressed as the relative telomere length. Telomere length differed significantly between the workers and controls (p<0.05). The results showed an indirect and significant association (r=-0.356, p=0.001) between age and telomere length in the workers. This study showed that the difference in telomere length shortening was statistically significant (p<0.05) between the workers and controls. It was concluded that occupational exposure acts as a risk factor to enhance telomere length shortening and accelerate ageing. Veza Između Skraćenja Telomera i Starenja Usled Profesionalne IzložEnosti Dužina telomera smatra se biomarkerom starenja i tokom ovog procesa rezultira skraćenjem. Ovo istraživanje predstavlja pokušaj određivanja relativne dužine telomera kod radnika u mehaničarskim radionicama. Skraćenje dužine telomera u ćelijama tokom profesionalne izloženosti izaziva ubrzano starenje. Genomska DNK izolovana je iz epitelnih ćelija unutrašnje strane obraza sakupljenih od 240 osoba, koji su činili dve grupe: 120 profesionalno izloženih radnika i 120 neizloženih kontrolnih subjekata. Dužina telomera izmerena je tehnikom PCR u realnom vremenu. Specifični prajmeri telomera (T) i gena prisutnih u jednoj kopiji (S) upotrebljeni su za izračunavanje relativne razmere T/S i izraženi kao relativna dužina telomera. Dužina telomera značajno se razlikovala između radnika i kontrolnih subjekata (p<0,05). Rezultati su pokazali da između starosti i dužine telomera kod radnika postoji indirektna i značajna povezanost (r=-0,356, p=0,001). Ova studija je pokazala da je razlika u skraćenju dužine telomera između radnika i kontrolnih subjekata bila statistički značajna (p<0,05). Zaključeno je da profesionalna izloženost predstavlja faktor rizika za znatnije skraćenje dužine telomera i ubrzano starenje.

Does GSTP1 polymorphism contribute to genetic damage caused by ageing and occupational exposure

Does GSTP1 Polymorphism Contribute to Genetic Damage Caused by Ageing and Occupational Exposure? The aim of our study was to see the effects of GSTP1 polymorphism on biomarkers of ageing, including micronuclei (MN), comet tail length, and relative telomere length in automobile repair workers, who are exposed to a broad spectrum of potential mutagens. The analysis was performed on buccal cells collected from occupationally exposed and non-exposed (control) subjects. Samples were analysed using cytogenetic and molecular methods, including restriction fragment length polymorphism (RFLP), MN test, comet assay, and real-time PCR. The results confirmed the DNA damaging effects of substances used in the mechanical workshops, but did not confirm the influence of GSTP1 gene polymorphism on DNA damage. However, further studies on both occupationally exposed and control populations are needed to understand the relationship between GSTP1 polymorphism and genome damage. Pridonosi li polimorfizam gena GSTP1 oštećenju genoma uzrokovanom starenjem i profesionalnom izloženosti? Na populaciji radnika zaposlenih u radionicama za popravak automobila koji su svakodnevno izloženi različitim vrstama potencijalnih mutagena istražili smo utjecaj polimorfizma gena GSTP1 na vrijednosti biomarkera starenja, uključujući pojavu mikronukleusa (MN), dužinu repa kometa te relativnu dužinu telomera. Analize su provedene na stanicama bukalne sluznice, skupljenim od izloženih ispitanika i odgovarajuće neizložene (kontrolne) populacije. Uzorci su analizirani primjenom citogenetičkih i molekularnobioloških metoda, uključujući polimorfizam restrikcijskih fragmenata na osnovi njihove duljine (engl. restriction fragment length polymorphism, RFLP), MN-test, komet-test, i lančanu reakciju polimerazom u stvarnom vremenu (engl. real-time PCR). Dobiveni nalazi potvrđuju da izloženost radnika mutagenima oštećuje njihovu DNA, ali nisu pokazali da polimorfizam gena GSTP1 značajno utječe na razinu oštećenja DNA. Zbog malog broja ispitanika uključenog u ovo istraživanje za bolje razumijevanje odnosa između polimorfizma gena GSTP1 i oštećenja DNA potrebna su daljnja istraživanja, i na profesionalno izloženim ispitanicima i na ispitanicima kontrolne populacije.

Assessment of the reliability of a novel self-sampling device for performing cervical sampling in Malaysia

BACKGROUND The participation of women in cervical cancer screening in Malaysia is low. Self-sampling might be able to overcome this problem.The aim of this study was to assess the reliability of self-sampling for cervical smear in our country. MATERIALS AND METHODS This cross-sectional study was conducted on 258 community dwelling women from urban and rural settings who participated in health campaigns. In order to reduce the sampling bias, half of the study population performed the self-sampling prior to the physician sampling while the other half performed the self-sampling after the physician sampling, randomly. Acquired samples were assessed for cytological changes as well as HPV DNA detection. RESULTS The mean age of the subjects was 40.4±11.3 years. The prevalence of abnormal cervical changes was 2.7%. High risk and low risk HPV genotypes were found in 4.0% and 2.7% of the subjects, respectively. A substantial agreement was observed between self-sampling and the physician obtained sampling in cytological diagnosis (k=0.62, 95%CI=0.50, 0.74), micro-organism detection (k=0.77, 95%CI=0.66, 0.88) and detection of hormonal status (k=0.75, 95%CI=0.65, 0.85) as well as detection of high risk (k=0.77, 95%CI=0.4, 0.98) and low risk (K=0.77, 95%CI=0.50, 0.92) HPV. Menopausal state was found to be related with 8.39 times more adequate cell specimens for cytology but 0.13 times less adequate cell specimens for virological assessment. CONCLUSIONS This study revealed that self-sampling has a good agreement with physician sampling in detecting HPV genotypes. Self-sampling can serve as a tool in HPV screening while it may be useful in detecting cytological abnormalities in Malaysia.

Comparative Assessment of a Self-sampling Device and Gynecologist Sampling for Cytology and HPV DNA Detection in a Rural and Low Resource Setting: Malaysian Experience

PURPOSE This study was conducted to assess the agreement and differences between cervical self-sampling with a Kato device (KSSD) and gynecologist sampling for Pap cytology and human papillomavirus DNA (HPV DNA) detection. MATERIALS AND METHODS Women underwent self-sampling followed by gynecologist sampling during screening at two primary health clinics. Pap cytology of cervical specimens was evaluated for specimen adequacy, presence of endocervical cells or transformation zone cells and cytological interpretation for cells abnormalities. Cervical specimens were also extracted and tested for HPV DNA detection. Positive HPV smears underwent gene sequencing and HPV genotyping by referring to the online NCBI gene bank. Results were compared between samplings by Kappa agreement and McNemar test. RESULTS For Pap specimen adequacy, KSSD showed 100% agreement with gynecologist sampling but had only 32.3% agreement for presence of endocervical cells. Both sampling showed 100% agreement with only 1 case detected HSIL favouring CIN2 for cytology result. HPV DNA detection showed 86.2%agreement (K=0.64, 95% CI 0.524-0.756, p=0.001) between samplings. KSSD and gynaecologist sampling identified high risk HPV in 17.3% and 23.9% respectively (p= 0.014). CONCLUSION The self-sampling using Kato device can serve as a tool in Pap cytology and HPV DNA detection in low resource settings in Malaysia. Self-sampling devices such as KSSD can be used as an alternative technique to gynaecologist sampling for cervical cancer screening among rural populations in Malaysia.

Single nucleotide polymorphisms (SNPs) of exons 3, 4, 5 and 9 of the matrix metalloproteinase 2 (MMP-2) gene in breast carcinoma of Malaysian women using high resolution melting analysis

Background Breast cancer metastasis is a factor contributing to high disease mortality. Single nucleotide polymorphism (SNP) of the matrix metalloproteinase 2 (MMP-2) gene, which controls the enzyme for degradation of macromolecules of the connective tissues and extracellular matrix, has been associated with cancer invasion and metastasis. Aim The aim was to detect SNPs of exons 3, 4, 5 and 9 of the MMP-2 gene in breast carcinoma of Malaysian women using high resolution melting analysis (HRM). Methods Genomic DNA was obtained of both tumour mass and normal adjacent tissue from 60 Malaysian breast cancer patients. SNP scanning involving all four exons of MMP-2 coding regions was performed by HRM analysis on Rotor-Gene 6000 real-time PCR. The PCR products presented with aberrant melting curve pattern were purified and sequenced. Sequence trace data obtained were analysed with Staden Package for mutation detection. Results The scanning of the MMP-2 exons detected 4 known SNPs in exon 5 (678G>C, 750C>T), intron 5 (832+12C>T) and exon 9 (1380G>A). No SNP was detected on exon 3 and 4. Five patients carried both copies of variant A SNP 1380G>A while only one patient carried homozygous variant T SNP 832+12C>T. Only one patient with homozygous variant T allele SNP 1380G>A and SNP 678G>C had poor outcome with extensive metastasis and failed chemotherapy. Conclusion A relationship between SNP of MMP-2 with disease progression and poor therapeutic response is a strong possibility. The utilisation of HRM, a sensitive and cost effective method for scanning the MMP-2 gene, is suggested on all exons to elucidate the involvement of SNPs in cancer progression.

Mucinous adenocarcinoma arising from chronic perianal fistula

Perianal mucinous adenocarcinoma is a rare tumour which may be associated with long‐standing chronic perianal sepsis. Early diagnosis is challenging and is based on a high index of clinical suspicion and specific histological features. Definitive treatment is surgical, in the form of an abdomino‐perineal resection. We hereby describe a case of a perianal mucinous adenocarcinoma arising from long‐standing recurrent perianal fistula and complement this with a brief review of the literature pertaining in particular to the management of this condition.