Aberrant Promoter Methylation at CpG Cytosines Induce the Upregulation of the E2F5 Gene in Breast Cancer / 한국유방암학회지

PURPOSE: The promoter methylation status of cell cycle regulatory genes plays a crucial role in the regulation of the eukaryotic cell cycle. CpG cytosines are actively subjected to methylation during tumorigenesis, resulting in gain/loss of function. E2F5 gene has growth repressive activities; various studies suggest its involvement in tumorigenesis. This study aims to investigate the epigenetic regulation of E2F5 in breast cancer to better understand tumor biology. METHODS: The promoter methylation status of 50 breast tumor tissues and adjacent normal control tissues was analyzed. mRNA expression was determined using SYBR® green quantitative polymerase chain reaction (PCR), and methylation-specific PCR was performed for bisulfite-modified genomic DNA using E2F5-specific primers to assess promoter methylation. Data was statistically analyzed. RESULTS: Significant (p<0.001) upregulation was observed in E2F5 expression among tumor tissues, relative to the control group. These samples were hypo-methylated at the E2F5 promoter region in the tumor tissues, compared to the control. Change in the methylation status (Δmeth) was significantly lower (p=0.022) in the tumor samples, indicating possible involvement in tumorigenesis. Patients at the postmenopausal stage showed higher methylation (75%) than those at the premenopausal stage (23.1%). Interestingly, methylation levels gradually increased from the early to the advanced stages of the disease (p<0.001), which suggests a putative role of E2F5 methylation in disease progression that can significantly modulate tumor biology at more advanced stage and at postmenopausal age (Pearson's r=0.99 and 0.86, respectively). Among tissues with different histological status, methylation frequency was higher in invasive lobular carcinoma (80.0%), followed by invasive ductal carcinoma (46.7%) and ductal carcinoma in situ (20.0%). CONCLUSION: Methylation is an important epigenetic factor that might be involved in the upregulation of E2F5 gene in tumor tissues, which can be used as a prognostic marker for breast cancer.

Uprising trend of research among medical students in Pakistan

Medical students these days are increasingly involved in the research projects, as well as article writing. The most common reason for increase in research activity is to improve the Curriculum Vitae for competitive USA and UK residencies. Furthermore few medical students want to do research at an early stage of medical career in order to get exposure to research for the postgraduate training purposes. Despite of all these interesting facts, there are various hurdles that medical students are facing, such as poor guidance of mentors, hardship during data collection, lack of time for research purpose due to study related barriers, rejections from journal, high fee charges from the reputable journals etc. all these factors demotivate medical students to pursue research

Reasons of patients transfer from peripheral teaching and non teaching hospitals to a surgical unit, lady reading hospital Peshawar: a pilot study

Objective: to determine the reasons of referral/transfer of patients from other health facilities to a surgical unit of Lady Reading Hospital Peshawar

Methodology: this is a descriptive cross-sectional study conducted in surgical unit "C" Lady Reading Hospital [LRH] Peshawar from February 28 to September 14, 2014, using consecutive non-purposive sampling technique. All patients admitted in our unit living outside the catchment area of Peshawar were included. Age, gender, address, reason for referral and perception of the patients about Lady Reading Hospital Peshawar was recorded on a proforma. Data was analyzed using SPSS version 20. Mean and standard deviation was calculated for quantitative output responses and frequency and percentages for qualitative output responses. Chi-square test was applied on the variables when compared and p value of <0.05 was considered significant

Results: total patients received in the study period of [six and half months] were 195.Out of these 75.4% were male. The mean age of patients was 37.2+ 17.49 years. The percentage admitted via emergency was 88.7%. Out of 195 patients, 147[75.4%] attended a health facility before coming to LRH. The reasons given for referral was lack of equipment, lack of staff, lack of ICU, lack of specialty, lack of out of hours services and seropositivity given in 88[59.8%], 74[50.3%],71[48.2%], 70[47.6%], 70 [47.6%] and 04 [2.7%] patients respectively.Dissatisfaction towards the local health facilities was shown by 137 [70.3%]

Conclusion: the peripheral health care system is being underutilized resulting in excessive burden on tertiary care health facilities. It is therefore important to address the problems and to implement a structured referral system

Surgical site infection; in-vitro anti-bacterial activity of different brands of ceftriaxone against bacteria commonly implicated in surgical site infection and their cost effectiveness

Ceftriaxone is used in wide range of day to day microbial infections in clinical practice3. Despite the incumbent drug regulating authority in Pakistan, there is scanty literature comparing the anti-microbial efficacy of different available br and s of ceftriaxone. Objectives: To know the in-vitro activity of various br and s of ceftriaxone against bacteria most commonly isolated from surgical site infection [SSI]. A comparison of five days cost of these br and s will also be done. Design: Experimental study. Period: Feb 2013 to Aug 2013 Setting: Surgical "C" unit Lady Reading Hospital [LRH] in collaboration with departments of pharmacology Khyber Girls Medical College [KGMC] and microbiology department of Lady Reading Hospital Peshawar. Material and Methods: Isolates of five bacteria i.e. Staphylococcus aureus, Proteus mirabilis, Escherischia coli, enterobacter Spp, and Klebsiella pneumoniae, found sensitive to ceftriaxone were grown on 50 slops each and the zone of inhibition was checked for each of the ten br and s of ceftriaxone. Results: The zones of inhibitions of different br and s of ceftriaxone against the above mentioned bacteria were not significantly different. The cost of therapy was significantly different for ten br and s. Conclusions: Various br and s of ceftriaxone of variable cost had no influence on their activity against bacteria involved in SSI.

Frequency of common bacteria and their antibiotic sensitivity in patients with symptomatic cholelithiasis

To determine the frequency of common bacteria and their antibiotic sensitivity in patients with symptomatic cholelithiasis This cross sectional descriptive study was conducted at Department of Surgery, Lady reading hospital, Peshawar for 6 months i.e., from December 2012 to May 2013. The total number of patients were 126. They presented with symptomatic cholelithiasis. Bile was aspirated preoperatively before cholecystectomy from gall bladder and was sent for culture and sensitivity test. The data was analyzed using Statistical Package for Social Sciences[SPSS] version 10.0. On culture and sensitivity test, 74 [58.73%] have positive growth while 52 [41.27%] have no growth. The most common bacteria was E. Coli isolated in 28 [22.22%] patients followed by Klebseilla in 22 [17.46%], Salmonellain 16 [12.70%] and Shigella in 8 [6.35%] patients. On culture and sensitivity test, all the 4 isolated bacteria showed sensitivity to Cefuroxime, Ceftriaxone and Ciprofloxacin in more than 50 % cases, while all the four bacteria showed resistance to amoxicillin in more than 50 % cases. The most common bacteria of symptomatic cholelithiasis are E. coli and Klebseilla followed by Salmonella and Shigella. These bacteria showed maximum sensitive to cefuroxime and ceftriaxone

Typhidot M and Diazo test vis-à-vis blood culture and Widal test in the early diagnosis of typhoid fever in children in a resource poor setting

OBJECTIVE: Typhoid fever is a major public health problem. A test which is simple, reliable and can be carried out in small laboratories is the need of the hour. We prospectively evaluated typhidot M and Diazo tests vis-à-vis blood culture and Widal test in children. METHODS: Patients aged 6 months to 12 years, having fever of more than four days duration with clinical suspicion of typhoid fever were enrolled. Patients in whom other diagnosis was made served as control. The tests under scrutiny were validated against blood culture and then all the four tests were evaluated among patients who presented in the first week of illness. RESULTS: Blood culture was positive in only 27.3 percent of the cases. Among these culture positive cases, typhidot M test had the highest sensitivity, specificity, PPV and NPV of 90 percent (95 percent CI = 74.4-96.5), 100 percent (95 percent CI = 90.1-100), 100 percent (95 percent CI = 87.5-100), and 92.1 percent (95 percent CI = 79.2-97.3) respectively. Diazo test ranked next with sensitivity, specificity, PPV and NPV of 86.7 percent (95 percent CI = 70.3-94.7), 85.7 percent (95 percent CI = 70.6-93.7), 83.9 percent (95 percent CI = 67.4-92.9), 88.2 percent (95 percent CI = 73.4-95.3) respectively. Among clinically suspected typhoid cases, the overall sensitivity, of blood culture, Widal, typhidot M, Diazo was 27.3 percent (95 percent CI = 19.8- 36.3), 64.6 percent (95 percent CI = 55.3-72.9), 89.1 percent (95 percent CI = 81.9-93.7), 80.9 percent (95 percent CI = 72.6-87.2) respectively. In the first week of illness, typhidot M showed the best sensitivity [86.2 percent (95 percent CI = 69.4-94.5)] followed by Diazo [79 percent (95 percent CI = 61.6-90.2)], Widal [41.4 percent (95 percent CI = 25.5-59.3)] and blood culture [31 percent (95 percent CI = 17.3-49.2)]. CONCLUSION: Both Typhidot M and Diazo are good screening tests for the diagnosis of typhoid fever. Typhidot M is superior to Diazo but the latter is more suitable to resource poor settings being economic and easy to perform.

Screening children with severe short stature for celiac disease using tissue transglutaminase.

Objective. To determine the prevalence of anti-tissue transglutaminase in children and adolescents with severe short stature (<-3 SD). Methods. All children in age group of 1-18 years having height less than -3 SD for their age and sex, were included. For each child age and sex matched healthy control (height more than -2 SD) was taken. The included subjects (study & control group) were subjected to anti tissue transglutaminase (tTG) (IgA) antibody assay estimation. Results. Of the 112 cases, 23 were tTG positive, giving a prevalence of 20.5% for seropositivity among cases of short stature while all the controls were seronegative for tTG. All the 23 had tTG values above 40 U/ml and 11 had values above 100 U/ ml. On univariate analysis we found that the presence of chronic diarrhea (OR = 2.55, 95%CI - 1.08-5.98), bulky stools (OR = 3.03, 95%CI - 1.52-6.05), hemoglobin < 7 gm/dl (OR = 3.12, 95%CI - 1.55 - 6.29) and more severe short stature (<-4 SD) (OR = 0.41, 95%CI - 0.17- 0.95) had significant association with the tTG positivity. On logistic regression analysis in all cases, hemoglobin < 7gm/dl (OR = 0.090, 95%CI = 0.024-0.342) and bulky stools (OR=0.212, 95%CI = 0.069-0.649) were significantly associated with tTG positivity. Conclusion. More than one fifth of all severe short stature are seropositive for tTG and the chances of seropositivity increases if severe anemia and bulky stool are also associated.