Investigation of Staphylococcus aureus, prevailing in the environment of Khyber Teaching Hospital, Peshawar, Pakistan

The hospital environment plays an important role in the spread of microorganisms, including multi drug resistant [MDR] strains. Patients can acquire Methicillin-Resistant Staphylococcus aureus [MRSA] which can reside in the clinical setup that are not cleaned and can spread through air droplets, bed clothing, and healthcare workers. The purpose of this study was to investigate the prevalence of S. aureus in the Khyber Teaching Hospital [KTH]. A total of 200 samples were collected from the floor, walls, air and inanimate objects in different wards of the KTH, during May 2012 to September 2012. These samples were screened for the recovery of S. aureus. Recovered organisms were subjected to susceptibility testing and investigated for the detection of various toxin and antibiotic resistance genes by Polymerase Chain Reaction [PCR]. A total of 64 samples yielded S. aureus, out of which 37 [57.81%] were proved as MRSA. No isolate was found resistant to Vancomycin, however 81.25% of the isolates were found susceptible to Linezolid and Amikacin. The susceptibility to Fusidic acid, Chloramphenicol, Rifampicin, Doxycycline and Meropenem was observed as 79.69%, 76.56%, 75.00, 73.44% and 68.75% respectively. The frequency of sea, seb and sec genes were 56.25%, 43.75% and 12.5% in the recovered isolates. Erm C was more prevalent [28.12 %] than the ermA and ermB. The prevalence of pvl in MRSA was 21.62 % which is less than 33.33% in the MSSA isolates. S. aureus and especially MRSA are frequently prevalent in the KTH. Therefore, every immune-compromised patient is prone to infections caused by S. aureus. This will lead to high morbidity/mortality rate, prolong hospital stay and add extra cost to the health system

Whole genome sequencing instead of whole exome sequencing is required to identify the genetic causes of polycystic ovary syndrome in Pakistani families

Background and Objective: Polycystic Ovary Syndrome [PCOS] is the major cause of infertility in females. PCOS is a complex and multifactorial disease, genetic and environmental factors being important predisposing factors. Diagnosis of PCOS is difficult due to the complexity of this disease; hence, better diagnostic tests are required to improve its management. Aim of the study was to elucidate the genetic causes of PCOS in three Pakistani families

Methods: Three Pakistani families segregating PCOS in an apparently autosomal recessive mode were recruited. Whole genome Single Nucleotide Polymorphism [SNP] genotyping and Whole Exome Sequencing [WES] were carried out to identify the candidate genes

Results: SNP genotypes data analyses identified multiple regions of homozygosity on different chromosomes. WES was performed in affected members of the family. Screening for pathogenic mutations in homozygous regions failed to detect any mutation/variant of interest

Conclusion: PCOS is multifactorial and complex disease so variants in the coding as well as in non-coding regions may be the genetic causes of the disease. To elucidate the genetic cause[s] of the PCOS, Whole Genome Sequencing [WGS] is recommended to cover both coding and non?coding regions of the genome

Evaluation of non-structural protein-1[NS1] positive patients of 2013 dengue outbreak in Khyber Pakhtunkhwa, Pakistan

Background and Objective: Dengue infection is an arthropod borne disease caused by Dengue virus in humans. Dengue virus infection has more potential to produce severe form of the disease with more severe symptoms. Proper diagnosis of dengue fever is very important for its safe management.The objective of this study was to evaluate the non structural protein-1 [NS1] positive parameter for identification of dengue fever by using ELISA from 2013 dengue outbreak in Khyber Pakhtunkhwa

Methods: It was a cross sectional study conducted among 384 patients tested for dengue admitted to different hospitals of Khyber Pakhtunkhwa April to December 2013 with symptoms related to classical dengue fever. Written informed consent was taken from 100 NS1 positive diagnosed patients, and 3 to 5 ml blood sample was collected for confirmation through ELISA testing. ELISA test for dengue IgG and IgM was performed two time in order to confirm the dengue cases. Data was entered and analyzed by using SPSS version 16

Result: The study performed on 100 NS1 positive samples of patients, admitted to hospitals with symptoms related to classical dengue fever, indicated that after performing the IgM and IgG capture ELISA test only 76 samples were actually found positive for dengue. The rest of the 24 samples were found negative for both IgM and IgG capture ELISAs. The study also revealed that 90.8 % patients had primary dengue infection and 35.5% patients had secondary dengue infection. Most patients were between the age of 10-20 years [26%], among them19.7% were having primary dengue infection. Among 10-20 years of age 50% female patients were false dengue patients

Conclusion: About 24 % NSI protein positive samples were found negative for both IgM and IgG capture ELISAs showed that NS1protein positivity does not confirm actual dengue infection

Diagnosis and phenotypic assessment of Pakistani Haemophilia B carriers

Objectives: 1: To assess the diagnostic utility of three polymorphisms [DdeI, XmnI and TaqI] and direct sequencing in haemophilia B [HB] carrier detection in Pakistani families. 2: To compare phenotypes of HB carriers with those of healthy females

Methods: The study was conducted from March 2014 till February 2016 at Khyber Medical University Peshawar and National Institute of Blood Diseases, Karachi. Individuals from HB families of Khyber Pakhtunkhwa [KP] and Federally Administered Tribal Areas [FATA] with known F9 mutation in the proband were enrolled into the study. FIX activity [FIX:C] levels were determined in all the participants. Bleeding scores [BS] and complete blood counts were performed in the female participants. Linkage analysis followed by targeted Sanger sequencing was carried out in all the study participants. Heterozygosity rate was determined for each polymorphism. Healthy females and the carrier groups were compared for bleeding phenotypes

Results: A total of 30 males and 48 females from 13 HB families were studied. The polymorphisms had a low heterozygosity rate. Direct sequencing determined the carrier status in all cases. The mean FIX:C was reduced whereas BS was raised in the carriers when compared with healthy females. A significant raise in white blood cells [WBCs] count was observed in the carriers

Conclusion: The three polymorphisms have a low heterozygosity rate in HB families from KP and FATA. Sanger sequencing is conclusive in determining carrier status in all the cases. FIX:C is low and BS is raised in the HB carriers in comparison to that of normal females. The mean WBCs count is significantly higher in the HB carriers than the normal females

Frequency of hepatitis B, hepatitis C and human immunodeficiency viruses in internally displaced persons of south Waziristan, Pakistan

Objectives: To investigate the burden of hepatitis B, hepatitis C and human immunodeficiency virus and to explore route of transmission in internally displaced persons of South Waziristan

Methodology: A cross sectional study was carried out in Gomal Medical Collage, Dera Ismail Khan; Mufti Mehmood Teaching Hospital Bannu and Basic Health Unit Zaferabad, Dera Ismail Khan. Total 300 internally displaced persons [IDPs] of South Waziristan who were undiagnosed and presented with non-specific symptoms were included in the study. The data was analyzed in SPSS version 16. Confidentiality of subjects was ensured

Results: In this study 300 IDPs were screened for HBsAg, anti-HCV and anti HIV antibodies. Out of which male were 190 and female were 110. More males were suffering from hepatitis B [10.3%] than hepatitis C [6.7%] or HIV [0.7%] as compared to females [3.3%], [5%] and [0.3%] respectively. A total of 5[1.7%] cases had history of previous surgery and 17[5.7%] had history of tooth extraction

Conclusion: The frequency of HBV was higher as compared to HCV and HIV infections. The frequency of all the three infections was comparatively more in males as compared to females. Most likely Most likely risk factors for acquistion of infection were unhygienic tooth extractions, unsafe blood transfusions and iatrogenic

Characterization of pathogens involved in ventilator associated pneumonia in surgical and medical intensive care units - A single center experience

In the present study 60 samples were collected from lower respiratory tract of patients suffering from Ventilator Associated Pneumonia [VAP] admitted in surgical and medical Intensive Care Units [ICUs] of Khyber Teaching Hospital, Peshawar, Pakistan. Recovered pathogens were characterized and their susceptibility pattern against commonly used antibacterial agents investigated. Most frequent bacterial pathogen found was methicillin- resistant Staphylococcus aureus [MRSA] [40%] followed by members of Enterobacteriaceae [22%; of which Escherichia coll [50%], Klebsiella pneumonia [30%], Enterobacter cloacae [10%] and Citrobacter freundii [10%], Pseudomonas aeruginosa 20% and Acinetobacter baumannii 18%. Majority of the specimens yielded polymicrobial growth [85.75% polymicrobial growth compared to 14.25% specimens yielding monomicrobial growth]. The susceptibility pattern showed that A. baumannii was the most resistant bacterial pathogen. Based on the results of susceptibility pattern obtained in the present study, combination of linezolid with meropenem and colistin has been found to be the best combination option for empirical therapy for VAP pathogens in this region

Modified acid fast staining: a better diagnostic tool in chronic diarrhoea due to cryptosporidiosis

Background: Cryptosporidium parvum is an emerging pathogen responsible for chronic diarrhoea in children and immuno-compromised individuals, especially AIDS patients. Currently, there is no effective therapeutic strategy for treating cryptosporidiosis, therefore control and supportive treatment of cryptosporidiosis depends upon rapid and accurate diagnosis of this infection. Methodology: A descriptive study was conducted in the Pathology Department of Khyber Medical College and Pathology Laboratory of Khyber Teaching Hospital over a period of one year March 2007–April 2008. A total of 200 stool samples were tested for the presence of C. parvum oocysts from children <5 years age suffering from diarrhoea for >5 days. Total and differential leukocyte count was determined to assess immune status of the patients. Modified Ziehl-Neelsen [Z-N] staining, a rapid, sensitive and easy test, was used successfully for the detection of C. parvum oocysts in stool specimen. Results: Cryptosporidium oocysts were found in 18 [9.0%] samples. Out of 18 positive cases, 13 [72.2%] children had lymphopenia hence their immune status was impaired. Infection was common in children between 1–24 months of age. Mean duration of diarrhoea was 11 months. Most of C. parvum infected children were consumers of well water [77.8%]. Conclusion: Cryptosporidiosis, although a self-limiting disease, rarely investigated routinely, can become chronic and life threatening in immuno-compromised individuals. Majority of affected patients are immunecompromised. Modified Z-N is a sensitive and rapid method which can explore the gravity of this infection even further if used routinely and may control morbidity and mortality associated with this infection. Keywords: Chronic diarrhoea, modified Ziehl-Neelsen staining, immuno-comprised

utility of fasting plasma glucose in predicting glycosylated hemoglobin in type 2 diabetes

Access to glycosylated hemoglobin [HbA1c] assays in clinical practice remains limited. We investigated the relationship of fasting plasma glucose and HbA1c to determine optimal glucose levels for predicting HbA1c. We retrospectively analyzed data on 2888 patients with type 2 diabetes mellitus aged 2:20 years using a linear regression of HbA1c against fasting plasma glucose. A receiver-operating characteristic analysis was used to determine optimal cut-points for fasting glucose in relation to HbA1c, area under the curve, sensitivity and specificity, and 95% confidence intervals [CI] for each cut-point. The mean [standard deviation] for the age of patients was 52 +/- 11.6 years. The average HbA1c was 8.9 +/- 2.46% and mean fasting plasma glucose was 10.1 +/- 3.62 mmol/L. The prevalence of HbA1c >/= 7.0%, and >6.5% was 76% and 82%, respectively. Overall, fasting plasma glucose and HbA1c were linearly correlated [r=0.62, P=0.001]. A fasting plasma glucose of >9.0 mmol/L predicted HbA1c 2:7.0% with an area under the curve = 0.807 [95% CI, 0. 0.794 to 0.821], while fasting plasma glucose >8.2 mmol/L predicted HbA1c >6.5%, with an area under the curve = 0.805 [95% CI, 0.791 to 0.818]. The sensitivity of both cut-points was 64.5% and 70.7% the specificity was 82.7% and 76.4%, the positive likelihood ratio was 3.73 and 2.99, and the positive predictive value was 92.2% and 93.2%, respectively. When HbA1c determination is not available, fasting plasma glucose levels may be used to identify patients with uncontrolled type 2 diabetes and initiate timely intensification of therapy to avoid long-term complications of diabetes

Immunization status against hepatitis B virus and determination of anti-HBS antibody titer in medical and dental students

This study was aimed to determine anti-HBs antibodies titer in medical and dental students to monitor success of hepatitis B vaccination. This study was conducted on students of Khyber Medical College and Khyber College of Dentistry, Peshawar. Serum samples from students who were vaccinated were tested for the determination of anti-HBs antibody titer using microparticle enzyme immunoassay [MEIA]. Out of 520 students surveyed, only 176 [33.84%] had either one injection or complete course of vaccine. None of them had checked their anti-HBs antibodies titer; 119 students [67.61%] had used Engerix-B, 41 [23.30%] students did not remember the brand of the vaccine while only 16 [9.09%] had used Heber-biovac HB. Twelve [6.8%] students tested were non-reactive for antibodies [titer <10.0 mIU/ml] while in 36 [20.5%] students antibodies titer was 10 and 100 mIU/ml respectively and 128 [72.7%] had antibodies titer more than 100 mIU/ml. Time lapsed between vaccination and detection of antibody titer was U - 6 years, 76 [43.1%] students had been tested after one year and 5 [2.8%] after 6 years and 41 [23.5%] students after 2 years. Only one third had their partial or complete course of their vaccination. Antibody titers were reactive in majority of students who had completed the course. Students entering medical and dental schools need to be better educated about hepatitis B and its vaccination

Antibacterial activity of nicotine and its nickel complex

Nicotine and its metal complex; Ni[II]-nicotine was isolated from leaves of Nicotiana tabacum using metal ions by the methods reported elsewhere. Their antibacterial activity was investigated using ten different species of Gram positive and Gram negative bacteria. For comparison, antibacterial activity was also tested for pure sample of nicotine and the metal salts used for complexation; Nickel[11] chloride, using the same set of bacterial strains under similar conditions. Nicotine showed no antimicrobial effect at lower dose level. On the contrary, at higher dose level, nicotine showed antibacterial activity against Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis, with a zone of inhibition of 14mm. Nickel[II] chloride was found to be effective against Aeromonas sobria and Staphylococcus aureus at lower dose level whereas it was effective against all the species of bacteria at higher dose level except Salmonella typhi and Shigella bovdii. Nickel[II] nicotine complex was ineffective against all the bacteria tested at lower dose level but effective against Aeromonas sobria, Vibrio cholerae and Bacillus subtilis

Antifungal activity of nicotine and its copper complex

Nicotine and its metal complex, Cu[11]-nicotine were isolated from leaves of Nicutiana tabacum using various metal ions by a techniques reported by Munir et al., 1994'. Antifungal activity of this complex was studied using 14 different species of fungi by comparing growth inhibition. Range of antifungal activity was compared with pure sample of nicotine and the metal salt used for complexation; copper[11] chloride, using the same species of fungi under similar conditions. Nicotine exhibited antifungal activity against 9 out of 14 selected species of which Copper[11] chloride was found to be effective against 11 out of 14 selected species. On the other hand, Copper[11]-nicotine was also found to be effective against 1 l out of 14 selected species of fungi tested