Evaluation of type 2 diabetes risk variants (alleles) in the Pashtun ethnic population of Pakistan / Journal of the ASEAN Federation of Endocrine Societies

Objective@#To evaluate the Type 2 Diabetes (T2D) risk variants in the Pashtun ethnic population of Khyber Pakhtunkhwa using nascent whole-exome sequencing (WES) to better understand the pathogenesis of this complex polygenic disorder.@*Methodology@#A total of 100 confirmed patients with T2D of Pashtun ethnicity were included in the study, DNA was extracted from whole blood samples, and paired-end libraries were prepared using the Illumina Nextera XT DNA library kit carefully following the manufacturer’s instructions. Illumina HiSeq 2000 was used to obtain sequences of the prepared libraries followed by bioinformatics data analysis.@*Results@#A total of n=11 pathogenic/likely pathogenic variants were reported in the CAP10, PAX4, IRS-2, NEUROD1, CDKL1 and WFS1. Among the reported variants CAP10/rs55878652 (c.1990-7T>C; p.Leu446Pro) and CAP10/rs2975766 (c.1996A>G; p.Ile666Val) identified were novel, and have not yet been reported for any disease in the database. The variants CAP10/rs7607759 (c.1510A>G, p.Thr504Ala), PAX4/rs712701 (c.962A>C; p.His321Pro), PAX4/ rs772936097 (c.748-3delT; p.Arg325Trp), IRS-2/rs1805097 (c.3170G>A; p.Gly1057Asp), NEUROD1/rs1801262 (c.133A>G; p.Thr45Ala), CDKL1/rs77152992 (c.1226C>T; p.Pro409Leu), WFS1/rs1801212 (c.997G>A; p.Val333Ile), WFS1/rs1801208 (c.1367G>A; p.Arg456His), and WFS1/rs734312 (c.1832G>A; p.Arg611His) are previously identified in other ethnic populations. Our study reconfirms the associations of these variants with T2D in the Pakistani Pashtun population.@*Conclusion@#In-silico analysis of exome sequencing data suggests a statistically substantial association of all (n=11) identified variants with T2D in the Pashtun ethnic population. This study may serve as a foundation for performing future molecular studies aimed at unraveling T2D associated genes.

Inadequate vitamin D level: association with low energy fractures of distal radius in young patients and its predictors in Karachi, Pakistan

Background:The objective of the study was to determine association of inadequacy of vitamin D level with low energy fractures of distal radius and its predictors in young patients in Karachi Pakistan. Methods:Cross-sectional study was conducted on 220 young patients, visited hospital with low energy fracture of distal radius.A questionnaire was designed and filled after taking consent includes details regarding age, gender, occupation, area of skin and sun exposure duration, dietary habits, type of clothing and residence used. Serum Vitamin D3 levels were determined and compared with serum calcium levels, serum phosphorus and alkaline phosphatase levels.Results:Among 220 patients ranging from 12-45 years, mean 28±23.33SD. Female were 172(78.18%). Mostly have history of fall 127(57.72%)and RTA 63(28.63). Sun exposure duration in majority of participant was 1-2 hour/day 132(60%). Variable coloured clothes used by majority participant 135(61.36%) and variable fabric 102(46.36%). 202(91.8%) patients had deficiency of vitamin D and correlated with duration of sunlight exposure significantly, also with exposure of large skin area, dietary consumption of vitamin D rich food and worn variable clothing colours. Serum phosphorus level and serum alkaline phosphatase level were negatively correlated with vitamin D significantly whereas positively correlated with serum calcium.Conclusions:Prevalence of vitamin D deficiency is very high in low energy fracture of distal radius in young populationand sun exposure duration found to be most common predictor of inadequate D levels. A national food fortification program and campaign of public awareness to increase sunlight exposure and increase intake of vitamin D rich food are urgently needed.

Efficacy and safety of sofosbuvir based antiviral therapy for chronic hepatitis C infection in patients with advanced chronic kidney disease

Background: Screening studies for hepatitis C have proved that it is more prevalent in patients with renal diseases. Chronic hepatitis C infection in patients with kidney disease not only accelerates renal deterioration but also adversely effects morbidity and mortality. Availability of direct acting antiviral drugs has revolutionized treatment of hepatitis C even in difficult patients. In advanced kidney diseases, selection of treatment is difficult. Aim of this study was to evaluate the efficacy and safety of Sofosbuvir based DAAs in patients with advanced CKD.Methods: In this Quasi experimental study, CHC patients with or without cirrhosis having advance CKD (eGFR <30 ml/min per 1.73 m2) and/or on dialysis were enrolled. End points of the study were documentation of SVR 12 or discontinuation of therapy. Different regimens of oral DAAs with or without Ribavirin were used.Results: 86 patients with a median age of 53 years were enrolled. 37 patients were on maintenance dialysis and 49 were not on dialysis with eGFR <30 ml/min per 1.73 m2. Virological response was 92.68% at the end of treatment and SVR was achieved by 90.2% twelve weeks after therapy. Insomnia 14%, headache 11% and anemia 7% were main dverse effects. Mean eGFR and creatinine before and after treatment remained the same. Only 2 patients relapsed, both were on dialysis thrice weekly.Conclusions: All Sofosbuvir based regimens used for the treatment of CHC in patients with end stage renal disease are effective and well tolerated. Close follow up is advised to monitor side effects.

Doppler ultrasound evaluation of cerebral blood flow in anaemia of chronic renal failure

Objective: To determine mean cerebral blood flow in anemia secondary to chronic renal failure by doppler ultrasound and to compare the results with data gathered from healthy control subjects. Study Design: Case control study. Place and Duration of Study: Combined Military Hospital, Lahore, from Jan 2011 to Jun 2011

Patients and Methods: A total of 60 subjects were included who were divided into two groups. Group I had 30 patients with anemia due to chronic renal failure while group II consisted of 30 healthy control subjects. Doppler for both groups was performed with a 7.5-MHz transducer of color doppler machine ALOKA SSD-5500 in a dimly lit room with a comfortable temperature [22°C-24°C] after an adaptation period of at least 15 minutes rest, in supine position. The internal carotid and vertebral arteries on both sides were examined in both groups and analyzed

Results: Patients with anemia due to chronic renal failure were found to have increased cerebral blood flow as compared to normal healthy individuals

Conclusion: Cerebral blood flow increases in patients with anemia due to chronic renal failure

Association of cholelithiasis with junk food and lipid profile in young women

Introduction: Cholelithiasis is a disease which is becoming quite prevalent in the society. Recent years have seen a dramatic change in the population afflicted by this disease along with its presentation. Changes in dietary habits with westernization of diet, sedentary life style and craving for fast food have resulted in a large number of female population belonging to younger age group [20-40 years] affected by gallstones

Objectives: To define the prevalence of gall stone disease in young female population, to highlight the importance of dietary factors and to define the role of lipid abnormalities

Study Design: Cross -sectional study

Place and Duration of Study: Surgical department of Allied Hospital, Faisalabad Medical University [FMU], Faisalabad from 20-6-17 to 20-10-17

Materials and Methods: A total of 50 patients and 30 controls were included in the study. Data was collected by a literature-based questionnaire regarding type of diet. Serum cholesterol and triglycerides were assayed by kit method. Abdominal ultrasonography was performed. The data obtained was subjected to statistical analysis using a SPSS version 17

Results: Results showed mean value for age 35.01 +/- 5.437 standard deviation. Out of 50 patients, 19 females were in the range of 25-35years [38.0 %] and 31 were in 36-45 years of age [62.0 %]. Out of 50 patients 16[32%] gave history of eating fast and fried food whereas 34[68%] patients never had taken any such food. Patients consuming soft drinks were 28[56%] and non-consumer patients were 22[44%].Patients having increased cholesterol were 7[14%].Normal level was observed in 43[86%] patients. Increased triglyceride level was found in 28[56%] while normal level in 22[44%] patients

Conclusion: The results indicated that the incidence of gallstones is increasing in younger females even up to 25 years. There is strong association of the disease with increasing age and junk food. No association was found with soft drink consumption. Also, no correlation was found with increased cholesterol and triglycerides

Manufacturer tolerance in slot height of 0.022-inch maxillary canine brackets

Orthodontic brackets manufacturer does not reveal their actual slot height. Tolerance if present in bracket slot can lead to poor three-dimensional control of teeth. This study was done to determine the manufacturer tolerance in slot height of maxillary canine brackets.Mesial and distal slots of one hundred and forty stainless steel maxillary canine brackets of seven commercial brands in 0.022-inch ["] slot were measured individually with leaf gauges of 0.01millime-ter [mm] thickness. Digital readout in inches of leaf gauges was attained by micrometer. Descriptive statistics were generated for bracket slot height and one sample T test was used to check whether significant tolerance exist in slot height. A p value < 0.05 was considered as significant.Smallest mean slot height of 0.0233+.0006 " was noted in mesial slot of Db orthodontics brackets while largest mean height of 0.0258+.0015 " were present in mesial slot of Mesal brackets. Maximum slot height of 0.028 " was noted in Precise and Sia brackets. Increase tolerance in brackets were noted in the range of 5.9 to 17.2%. A p value < 0.05 was found in all the slot heights under study showing significant difference from acceptable tolerance.All the bracket series have significant oversized slot height on both mesial and distal aspect. Eu-ropean orthodontic brackets were closer to standards than brackets manufactured in USA

Acinetobacter Spp: Resistance and therapeutic decisions at the turn of the novel millennium

This study was planned to evaluate sample wise isolation and antimicrobial resistant trends of Acinetobacter spp in different departments of a tertiary care hospital. This was a transversal descriptive study, carried out in the clinical microbiology laboratory of the Allama Iqbal Medical College/ Jinnah Hospital, Lahore, Pakistan, during the period of January 2015 to December 2016. Every clinical specimen was processed for bacterial culture and antimicrobial susceptibly testing. A total of 3590 [2015=1780, 2016=1810] clinical specimens were processed. Of the total, only 54.7% were gram-negative, among these Acinetobacter spp were isolated from 10.1% and 16.5% samples respectively in 2015-16 with an overall rate of 24.3%. The highest occurrence of Acinetobacter spp isolates was reported from Intensive care units [ICU] [54%] followed by surgical units [25%] and medical units [16%]. It is noteworthy that ICU and internal medicine showed the highest resistance rates, whereas, lower resistance rate was observed for the outdoor patients [OPD]. Although collistin showed 0% resistant while ceftriaxone, ciprofloxacin, gentamicin, and tigecycline showed 90%, 68%, 66%, 66% and 62% resistance against Acinetobacter spp. respectively. An alarming increase in the resistance rate of meropenem, cefoperazone/sulbactam, piperacillin/ tazobactam, ciprofloxacin, and imipenem was observed from the year 2015 to 2016. This startling resistance acquired by Acinetobacter spp. within a period of one year, represent very limited therapeutic options left for the infections caused by Acinetobacter spp. Unavailability of effective drugs and limited therapeutic options enforce the health care practitioners to prescribe expensive and broad range antibiotics, which may cause harm to the patient. Therefore, it is need of an hour to better understand the antimicrobial patterns and optimize antimicrobial prescription policies for the control of multidrug-resistant Acinetobacter spp

Bugged mobile phones of health care providers could transmit more than just a call

Objective: To evaluate contamination of mobile phones from healthcare workers with nosocomial pathogens

Study Design: Cross sectional study

Place and Duration of Study: Microbiology Section, Pathology Department, Allama Iqbal Medical College, Jinnah Hospital Lahore, from Jul 2016 to Feb 2017

Material and Methods: A total of 257 culture swabs were collected from the mobile phones of HCWs of Jinnah Hospital, Lahore, every samples was processed for bacterial culture following standard protocols. Organisms were identified by colonial morphology, gram staining, and with the help of API 20NE and API 20E

Results: Out of 257 mobile phones, 66 percent [n=169] were contaminated with any 01/>01 bacteria. Most prominent pathogen isolated was coagulase negative staphyloccoi [CoNS] followed by Staphylococcus aureus 34.8 percent and 24 percent respectively. Other microorganisms identified, were Acinetobacter species [12.6 percent] n=29, Bacillus species 10.4 percent [n=24], Enterococcus species 4.8 percent [n=11], Escherichia coli 3.5 percent [n=8], Micrococcus species 3.5 percent [n=8], Diphtheroids 2.5 percent [n=5] Klebsiella species 1.7 percent [n=4], Aspergillus species 1.7 percent [n=4] and Pseudomonas aeruginosa 1.0 percent [n=2.0]. Out of 55 Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus [MRSA] were 36.0 percent [n=20] and methicillin-sensitive Staphylococcus aureus [MSSA] were 64.0 percent [n=35]

Conclusion: In the hospital setting mobile phones should be regularly decontaminated. Moreover, utmost emphasis needs to be paid to hand washing practices among HCWs

Pseudomonas aeruginosa: evaluation of pathogen burden and drug-resistance trends in a tertiary care hospital

Objective: To evaluate the pathogen burden and antibiotic-resistance trends of Pseudomonas aeruginosa among hospitalised patients at a tertiary care hospital. Study Design: Retrospective, hospital record-based, cross-sectional study. Place and Duration of Study: Microbiology Laboratory, Allama Iqbal Medical College/Jinnah Hospital, Lahore, from January 2014 to December 2016

Methodology: A total of 5,960 samples were collected from clinically suspected cases of bacterial infections, admitted to the hospital. Microbial identification and antibiotic susceptibility pattern were carried out and analysed

Results: Out of a total of 5,960 samples, Pseudomonas aeruginosa was isolated from 1,268 [21.2%] specimens. Department-wise isolation rate was n=600 [42.9%], n=268 [15.4%], n=201 [12.6%], and n=199 [16.0%] from intensive care unit [ICU], surgical units, medical units, and Gynae wards, respectively [p<0.0001]. Sample-wise isolation rate was, wound swabs n=448 [35%], urine n=356 [28%], sputum n=187 [14 %], tracheal aspirate n=127 [10%], blood n=99 [7%], and broncho-alveolar lavage n=51 [4%] [p<0.0001]. Drug-resistance pattern showed low rates for carbapenems [meropenem n=440 [35%], Imipenem n=436 [34%] and beta-lactam + beta-lactamase inhibitor combination [piperacillin+ tazobactam n=437 [34%] while alarming rates were observed for cephalosporins [ceftazidime n=716 [56%], fluoroquinolones [ciprofloxacin n=690 [54%], cefoperazone+sulbactam n=685 [54%], aminoglycosides [gentamicin, n=669 [53%], amikacin n=608 [48%], and monobactams [aztreonam n=666 [52%]. Decreasing trend was observed only for amikacin 63% to 37%, aztreonam showed similar pattern throughout, while there was an increasing trend of drug resistance in all groups of antibiotics

Conclusion: Emerging drug-resistant strains of Pseudomonas aeruginosa are probably linked to the injudicious use of antibiotics, leading to ineffective empirical therapy. Therefore, we suggest that culture and antimicrobial susceptibility testing should be done for targeted antimicrobial therapy against Pseudomonas aeruginosa

False negativity of ziehl-neelsen smear microscopy: is the scale-up the worth it in developing countries?

Objective: To evaluate the false negative results of Ziehl-Neelsen [ZN] smear microscopy

Study Design: Descriptive study

Place and Duration of Study: Mycobacteriology Laboratory, Allama Iqbal Medical College [AIMC] and Jinnah Hospital, Lahore [JHL], Pakistan, from February 2014 to August 2016

Methodology: A total of 3,951 [pulmonary 2,773 and extra-pulmonary 1,178] samples were collected from strong TB suspected patients attending JHL Lahore. Follow-up cases were excluded. Every specimen was processed for ZN smear microscopy, Lowenstein Jensen [LJ] culture. SPSS 21.0 was used; false negative and positive results of ZN smear were calculated keeping LJ culture as gold standard

Results: Out of total 3,951 samples, sputum was most freqently found pulmonary sample 48.4% [n=1915], extrapulmonary samples, pleural fluid and pus samples were most commonly observed samples 12.0% [n=476] and 8.3% [n=329], respectively. Overall false negativity was 23.1% [pulmonary=19.6%, extra-pulmonary=29.2%] [p<0.001], Maximum false negative results were observed in pericardial, synovial, pleural fluids, and pus samples as 40.0%, 38.0%, 33.0% and 32.0%, respectively

Conclusion: ZN smear microscopy is not a very efficient tool in case of patients with the low mycobacterial load. Therefore, National TB Control programs should consider extending their diagnostic approaches from ZN microscopy to more advanced techniques

Supracondylar humerus fractures- experience at a forward treatment centre in Kashmir

Objective: To assess the results of open reduction and internal fixation with K-wires for displaced Supracondylar humerus fractures in children in an isolated forward treatment centre [FTC] in Neelum valley. Study Design: Cross-sectional descriptive series. Place and Duration of Study: This study was carried out at a FTC which is located in Tehsil Headquarters Hospital Kel valley from Dec 2013 to Feb 2015

Material and Methods: It was a cross sectional descriptive study. Data was collected form 24 patients who came to FTC Kel with displaced supracondylar fracture from Dec 2013 to Feb 2015. They were assessed for age, sex, etiology, time of surgery, pattern of fracture, hospital stay, infection and range of motion

Results: In our study majority of patients were found to be between 5-6 years of age [66%]. The majority of the patients were males [83%]. The commonest cause was fall from mountain tracks [62%]. In our study we had twenty [83%] patients who had grade III displaced fracture. Ninety one percent underwent operation on the same day as the occurrence of injury [91%] patients were discharged on 3[rd] post op day. Twelve percent Patients had pin track infection and [4%] patients had iatrogenic nerve palsy which recovered spontaneously. Four percent patients developed superficial wound infection. There was loss of range of motion in six patients ranging from 0-5 degrees. Functional results based upon Flynn's grading system showed that we had 75% satisfactory result

Conclusion: We concluded that open reduction and internal fixation with k-wires at a remote station without the facility of image intensifier was safe and effective provided that the surgeon is confident in this procedure

Comparison of transversus abdominis plane block versus local infiltration of wound with bupivacine after appendisectomy

Objective: To compare the effect of transverses abdominis plane block verses local infiltration of wound with Bupivacaine to assess frequency of analgesic requirement for pain relieve during 15 hours after appendisectomy. Study Design: Randomized controlled trial. Place and Duration of Study: Combined Military Hospital Peshawar, from Apr 2016 to Oct 2016

Results: Both the groups were comparable with to assess frequency of analgesic for pain relieve after TAP Block Vs local infiltration with bupivacaine. In group A the frequency of analgesic requirement was less as compared to group B [p<0.05]

Material and Methods: Ninety two patients fulfilling the inclusion criteria were included by consecutive sampling technique for this study and divided into two groups of 46 each. Patients of group A were administered transverse abdominis plane [TAP] block via ultrasound guide. Patients in group B received local infiltration with Bupivacaine. Frequency of analgesic requirement for pain relieve were assessed by anaesthetist during 15 hours after appendisectomy

Conclusion: Frequency of analgesic requirement was less in TAP block as compared to local infiltration of wound with bupivacaine during 15 hours after appendisectomy

Comparison of central corneal thickness measurements with the galilei dual scheimpflug analyzer and ultrasound pachymetry

Objective: To determine the correlation between mean central corneal thickness taken with Galilei dual Scheimpflug Analyzer and Applanation Ultrasound Pachymetry

Study Design: Descriptive cross sectional study

Place and Duration of Study: Armed Forces Institute of Ophthalmology Rawalpindi, from Jul 2013 to Jan 2014

Material and Methods: Central corneal thickness was measured in 100 eyes of 50 patients. First three readings were taken with Galilei dual Scheimpflug analyzer, with a gap of 1 minute. Then three readings were taken with ultrasound pachymetry after applying topical 0.5% proparacaine [Alcain]. The mean of the three readings was used for the analysis

Results: For right eye the mean central corneal thickness measured by the Galilei dual Scheimpflug analyzer and Ultrasound pachymetry was 544.06?m +/- 27.36 and 546.88?m +/- 27.71 respectively, and for left eye it was 544.72?m +/- 25.47 and 546.52?m +/- 26.15 respectively. There was a strong and positive correlation between the two instruments [r=0.969, p=0.000 for right eye and r=0.956, p=0.000 for left eye]

Conclusions: The pachymetry readings with GSA showed strong and positive correlation with those of US pachymetry. So GSA may be considered as an alternative to US Pachymetry, thus avoiding operator-dependent errors, patient discomfort and other disadvantages

Left atrial to left ventricular end diastolic diameter ratio in left ventricular diastolic dysfunction

Background: Diastolic dysfunction [DD] is difficult to diagnose and characterize despite many techniques. It is known that left atrial diameter [LAD] increases and left ventricular internal end diastolic diameter [LVIDD] remains the same in moderate to severe diastolic dysfunction [grade II and above] but a ratio between these two [LAD/LVIDD] has never been used to assess diastolic dysfunction. This study aims to analyze the relationship between the ratio, left atrial diameter/ left ventricular internal end diastolic diameter [LAD/LVIDD] and diastolic dysfunction [grade II and above]. This study also aims to give a statistical model based upon this relationship that is able to help in diagnosing diastolic dysfunction

Methods: This case control analytical study was carried out at the Department of Cardiology, Holy Family Hospital, Rawalpindi, from Jan 2014 to Jan 2016. On the basis of echocardiographic findings, 226 patients were identified as normal healthy control group whereas 226 patients were identified as having diastolic dysfunction of grade II and above and were categorised as disease group. Doppler diastolic function parameters were used to assess diastolic function in all of these 452 patients. Appropriate statistical methods were then applied on the data by using IBM statistical package for social sciences [SPSS]

Results: LAD/LVIDD was correlated to diastolic dysfunction at r=0.858 [p<0.01]. A binary logistic regression model using age, sex and LAD/LVIDD gave an accuracy of 96.9% in diagnosing diastolic dysfunction. LAD/LVIDD in the model had an odds ratio of 2.678?1024. A Receiver Operation Characteristic curve between diastolic dysfunction and LAD/LVIDD yielded a specificity of 93.8% and sensitivity of 99.1% at the cut-off value of 0.7268

Conclusion: A new parameter, left atrial diameter [LAD] to left ventricular internal end diastolic diameter [LVIDD] ratio [LAD/LVIDD], is significantly correlated to diastolic dysfunction [grade II and above]. It is also a significant predictor for diastolic dysfunction

Evaluation of hepatotoxic and nephrotoxic effects of piroxicam sulfonated derivatives

Background: Pain management requires new pharmacotherapy with good efficacy and less side effects. Piroxicam is used routinely in clinical practice but it is associated with side effects. To minimize the chances of adverse effects, sulfonated piroxicam derivatives [SPD] have been introduced. We sought to find hepatotoxic and nephrotoxic effects of SPD in Albino rats

Methods: An experimental study on SPD was carried out at the Institute of Basic Medical Sciences, Khyber Medical University, Peshawar. Healthy 24 albino rats were divided into 5 groups. One control group and four experimental groups [compound I and II, each with a dose of 10mg/kg and 20mg/kg] received treatment for 7 days. Liver function tests [LFTs], renal function tests [RFTs] and histology of liver and kidney specimens was performed after culling rats. The difference between median values of samples was assessed using Kruskal-Wallis test with post-hoc [for LFTs and RFTs]. SPSS-21 was used for all statistical analysis and p?0.05 was considered statistically significant

Results: The alanine aminotransferase [ALT] values were significantly high in the 20 mg/Kg group than control for both compounds [p=0.03, p=0.001 respectively]. The aspartate aminotransferase [AST] values were significantly high in the 10 mg/Kg and 20 mg/Kg group than control for compounds II [p=0.01, p=0.0001 respectively]. The alkaline phosphatase [ALP] values were significantly high in the 20 mg/Kg group than control for compounds II [p=0.002]. The blood urea values were significantly high in the 20 mg/Kg group than control for compounds II [p=0.008]. The mean final score of liver injury in all experimental groups [mean range 5-7] was less suggesting that the damage in liver was less pronounced. Renal injury was more pronounced in the 20 mg/Kg dose for both compound I and compound II [mean score 7] compared to 10 mg/Kg dose [mean score 4]

Conclusion: Piroxicam sulfonated derivatives can cause focal changes in liver and kidney which might be reversible. The changes are less pronounced for compound I with a low dose

Rapid screening of pyogenic Staphylococcus aureus for confirmation of genus and species, methicillin resistance and virulence factors by using two novel multiplex PCR

Objectives: Emergence of methicillin resistant Staphylococcus aureus [MRSA] is a major medical problem of current era. These bacteria are resistant to most drugs and rapid diagnosis can provide a clear guideline to clinicians. They possess specific virulence factors and relevant information can be very useful. We designed this study to develop multiplex PCRs to provide rapid information

Methods: We studied 60 Staphylococcus aureus isolates and detected methicillin resistance by cefoxitin sensitivity and targeting of mecA gene. After initial studies with uniplex PCRs we optimized two multiplex PCRs with highly reproducible results. The first multiplex PCR was developed to confirm genus, species and methicillin resistance simultaneously, and the second multiplex PCR was for screening of virulence factors

Results: We found 38.33% isolates as methicillin resistant, a-toxin, the major cytotoxic factor, was detected in 40% whereas 6-hemolysin was found in 25% cases. Panton Valentine leucocidin was detected in 8.33% and toxic shock syndrome toxin in5% cases. The results of uniplex and multiplex PCRs were highly compatible

Conclusions: These two multiplex PCRs when run simultaneously can provide vital information about methicillin resistance and virulence status of the isolate within a few hours as compared to several days needed by routine procedures

Congenital Hypothyroidism in Neonates of a Tertiary Care Hospital

Objective: To determine neonatal congenital hypothyroidism among neonates born in a tertiary care hospital of Lahore Pakistan

Methods: This cross-sectional study was carried out at Pathology Department of Allama Iqbal Medical College, Lahore in collaboration with Pediatrics and Gynecology and Obstetrics Department, Jinnah Hospital, Lahore Pakistan. A total of 770 babies were included in this study, both male and female. About 2 ml venous blood samples were collected aseptically from the neonates in sterile clotted tube. Serum was separated and serum TSH was determined by ELISA method

Results: Out of total 770 neonates, 48.9% were female and 51.0% were males with the ratio of 1:1.04. Neonatal congenital hypothyroidisim [TSH, >30 mlU/L], was observed in 0.4% [Frequency, 1:257] nenates, with the incidence rate of 1:257. Female to male ratio of hypothyroid neonates was 2:1. The mode of delivery vise distribution showed, among n=251 neonates born by normal delivery, only a single case of hypothyroidism was detected, and among n=519 neonates delivered by cesarean section, only two neonates were belong to hypothyroidism

Conclusion: The frequency of Congenital Hypothyroidism is notably higher in pediatric community than reported in most other countries. This result emphasizes the necessity of a nationwide screening program

Rheumatoid arthritis therapies: present and future

Rheumatoid arthritis [RA] is an autoimmune disorder characterized by inflammation of synovium, cartilage damage and co-occurring of various other disorders. Significant improvement has been achieved in RA therapeutics in last two decades. However, newer, more efficient and more cost-effective therapeutic applications are still needed to be developed. Current therapies in RA are mostly acting to restrict inflammation. Non steroidal anti inflammatory drugs [NSAIDs] and disease modifying antirheumatic drugs [DMARDs] are conventionally used in RA therapy. Studies showed higher efficacy of anticyfokine therapy such as anti-IL-6 and anti-TNF. Rituximab, targeting B cells, after establishing its potential was approved to be used in combination with methotrexate in 2006. Abatacept, CLTA-4-IgGl, has been developed to block CD28 and CD80 or CD86 interaction leading to the termination T cell activation. Molecular inhibitors are relatively new in RA therapeutics such as tocilizumab, tofacitinib and baricitinib. PDA has approved tofacitinib to be used as a treatment for moderate and severe RA. Future holds promising therapeutic options based on numerous studies. IL-12, IL-17 and IL-23 are the targets of future anticytokine therapies. Several lymphocyte targeting agents including ofatumumab, ocrelizumab and veltuzumab have been developed and are currently in phase II and phase III clinical trials. There is a vast range of potential targets in RA enabling us to expand the therapeutic options over the next decade

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.

Comparison of intraocular pressure measurement by goldmann applanation tonometry and pneumotonometry in healthy subjects and patients with primary open angle glaucoma

Objective:To evaluate the role of non-contact Pneumo-tonometry [PT] by comparing intraocular pressure [IOP] measurements with Goldmann Applanation Tonometry [GAT] and PT in healthy subjects, and patients with Primary Open Angle Glaucoma [POAG]

Materials and Methods: This prospective comparative study was conducted at PNS Shifa Naval Hospital, Karachi from January 2016 to June 2016. A total of 360 eyes of 180 patients were included, and IOP measurement was done using GAT and PT methods. The difference in readings between the two methods was calculated and analysed using SPSS version 17

Results: A total of 360 eyes were analysed. 90 healthy subjects [180 eyes] and 90 patients with diagnosed POAG [180 eyes] were included. Mean IOP measured in all eyes using GAT and PT was 16.32 +/- 5.31 mm of Hg and 18.16 +/- 6.76 mm of Hg respectively. Mean difference in IOP was 2.31 +/- 1.89 mmHg between two methods, with 278 [77.22%] eyes showing higher IOP reading than those measured using GAT. The difference of readings between two methods was statistically significant. The patients with POAG were divided into three groups. Group 1 had IOP less than 15 mm Hg, Group 2 had IOP between 15-25 mm Hg and Group 3 had IOP greater than 25 mm Hg. Mean difference in IOP between two methods was 1.38 +/- 1.85, 2.29 +/- 2.72 and 3.05 +/- 2.68 mm Hg in Group 1,2 and 3 respectively. The mean difference of IOP measurement using GAT and PT in these groups was statistically significant [P<0.001]

Conclusion: IOP measurement using PT overestimates the recorded IOP as compared to GAT, and the difference is more pronounced in eyes with higher than normal IOP