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Objective: To study the genomic organization of vancomycin resistance in a local isolate of vancomycin resistant Staphylococcus aureus (VRSA)., Study Design: Experimental study., Place and Duration of Study: Department of Microbiology, University of Karachi, January 2008 through December 2010., Methodology: A vancomycin-resistant Staphylococcus aureus (VRSA-CP2) isolate (MIC 16 μg/ml) was isolated from a local hospital of Karachi. Species identification was confirmed by Gram staining, standard biochemical tests and PCR amplification of the nuc gene. The vancomycin MIC was re-confirmed by E-test. For the genetic determination of vancomycin resistance, in-vitro amplification of vanA cassette was performed by using plasmid DNA of CP2, CP2's transformant as template on MWG Thermo-Cycler. Amplified products of vanR, vanS, vanH, vanA, vanY, orf2, orf1D, orf2E, orf-Rev and IS element genes were subjected to Sanger's electrophoresis based sequence determination using specific primers. The Basic Local Alignment Search Tool (BLAST) algorithm was used to identify sequences in GenBank with similarities to the vanA cassette genes., Results: The vancomycin-resistant isolate CP2 was found to be resistant to oxacillin, chloramphenicol, erythromycin, rifampicin, gentamicin, tetracycline and ciprofloxacin, as well. The isolate CP2 revealed four bands: one of large molecular size ~56.4 kb and three of small size ~6.5 kb, ~6.1 kb and ~1.5 kb by agarose gel electrophoresis indicating the presence of 3 plasmids. The plasmid DNA of isolate CP2 was analyzed by PCR for the presence of the van cassettes with each of the vanA , vanB and vanC specific primers. It carried vanA cassette, which comprises of vanR, vanS, vanH, vanA, vanY, and orf2. The vanA cassette of isolate CP2 also carried an insertion element (IS). However, it did not show the PCR product for orf1. Vancomycin resistance was successfully transferred from the donor CP2 to a vancomycin-sensitive recipient S. aureus. The MIC of vancomycin for the transformant was 16 μg/ml, similar to the parent isolate CP2. Nucleotide sequencing of the PCR product showed similarity with van genes of enterococci and other VRSA reported from different parts of the world., Conclusion: Sequence of vanA cassette of CP2 showed partial homology with vancomycin resistant enterococci, VRSA vanA cassette element recorded in gene bank NCBI.

Adsorption is an environmental friendly process for removal and/or recovery of heavy metals from wastewater. In recent years, it has been substantiated as a popular technique to treat industrial waste effluents, with significant advantages. In this work, batchwise removal of chromium (III) ions from water by Polyalthia longifolia leaves was studied as a function of adsorbent dose, pH, contact time, and agitation speed. Surface characteristics of the leaves were evaluated by recording IR spectra. The Langmuir, Freundlich, and Temkin adsorption isotherms were employed to explain the sorption process. It was found that one gram of leaves can remove 1.87 mg of trivalent chromium when working at pH 3.0. It has been concluded that Polyalthia longifolia leaves can be used as cost-effective and benign adsorbents for removal of Cr(III) ions from wastewater.

Biofilms are means of protection to bacteria against antibiotics and antibodies. Catheters and others tube devices used by patients are prone to accumulation of thick layers of biofilms as hiding place for etiologic agents, resulting in substantial morbidity and mortality. Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired infections. Vancomycin remains the only treatment of choice for MRSA infections. In the present study a vancomycin resistant S. aureus (VRSA) (Labeled as CP2) was isolated from the blood of a post-operative cardiac patient. It harbors a plasmid which carry vanA gene and exhibited low-level vancomycin resistance (MIC 16 μg/ml), high level of oxacillin/methicillin resistance (MIC 500 μg/ml) and was sensitive to teicoplanin. CP2 also found to carry icaA gene on its chromosome. This strain exhibited resistance to triton-X100 induced autolysis under sub-inhibitory concentration of vancomycin and produced some extracellular matrix material that surrounding the cells. These characteristic features have warranted us to study the biofilm formation by CP2 on biomedical indwellings in presence of vancomycin and oxacillin. Our findings suggest that sub-lethal dose of vancomycin induced the biofilm formation by CP2 on nylon and silicon indwellings whereas oxacillin facilitated the biofilm formation on glass surfaces exclusively. This implicates that not only the antibiotics but also the indwelling material influences biofilm formation. Therefore, these implants serve as potential surfaces for bacterial adhesion that lead to biofilm formation, thus provide hiding places for pathogens from the actions of antimicrobials., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The in vitro antiproliferative and antioxidant activities of the aqueous, chloroform and methanol extracts of Muntingia calabura leaves were determined in the present study. Assessed using the 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay, the aqueous and methanol extracts of M. calabura inhibited the proliferation of MCF-7, HeLa, HT-29, HL-60 and K-562 cancer cells while the chloroform extract only inhibited the proliferation of MCF-7, HeLa, HL-60 and K-562 cancer cells. Interestingly, all extracts of M. calabura, which failed to inhibit the MDA-MB-231 cells proliferation, did not inhibit the proliferation of 3T3 (normal) cells, indicating its safety. All extracts (20, 100 and 500 μg/ml) were found to possess antioxidant activity when tested using the DPPH radical scavenging and superoxide scavenging assays with the methanol, followed by the aqueous and chloroform, extract exhibiting the highest antioxidant activity in both assays. The total phenolic content for the aqueous, methanol and chloroform extracts were 2970.4 ± 6.6, 1279.9 ± 6.1 and 2978.1 ± 4.3 mg/100 g gallic acid, respectively. In conclusion, the M. calabura leaves possess potential antiproliferative and antioxidant activities that could be attributed to its high content of phenolic compounds, and thus, needs to be further explored.

This study explores the bereavement process and continuing bond in Pakistani Muslims with the focus on how culture and religion influence these processes. Ten participants were interviewed and their transcribed interviews were analyzed using a grounded theory approach. Three main domains were identified from the narratives expressed by the participants: death and the process of grieving, continuing the link with the deceased, and influencing agents. The findings indicated that Pakistani Muslims maintained their link with the deceased through cultural and religious rituals, such as performing prayers, reciting holy verses, talking and dreaming about the deceased, doing charity, visiting graves, and arranging communal gatherings. The prime purpose of many of these practices was the forgiveness of the deceased. Grief reactions seemed to be determined by the nature of death, prior relationships with the deceased, reaction of society and gender of the bereaved. Religion provided a strong basis for coping and adjustment of the bereaved, through rationalizing and accepting the death. This study has important implications for counselors and family therapists who can use religious affliations to reduce the impact of loss and complicated bereavement.

Chondrosarcomas are malignant cartilage tumours. They are poorly responsive to chemotherapy and radiotherapy. Treatment is usually limited to surgical resection; however, survival of patients with high-grade chondrosarcoma is poor, even with wide surgical resection. Induction of apoptosis in chondrosarcoma cells, either directly or by enhancement of the response to chemotherapeutic drugs and radiation, may be a route by which outcome can be improved. In this article, we review potential molecular targets that regulate chondrocyte apoptosis and discuss the experimental evidence for their utility., (© 2010 The Authors. International Journal of Experimental Pathology © 2010 International Journal of Experimental Pathology.)

A Vancomycin intermediate resistant strain of Staphylococcus aureus (S. aureus) labeled as CP2 (MIC 16 microg/ml) was isolated from an in-patient of local Cardiac Hospital of Karachi. CP2 showed typical characters of Vancomycin intermediate resistant S. aureus (VISA) i.e. high level of oxacillin resistance, thickened cell wall with rough surface and reduced autolytic activities associated with murein hydrolase (MH) enzyme. This strain may have acquired vancomycin resistance due to long term exposure to antibiotic during the treatment of the patient. Therefore, it implicates the importance of monitoring the usage and also to control of the abuse of antibiotics for prevention of any further proliferation of this type of notorious bugs.

Squamous cell carcinoma of the oral cavity (OCSCC) is the most frequently observed form of head-and-neck cancer in Southeast Asia and is the sixth most common cancer worldwide. Most cases of this preventable disease are caused by alcohol consumption, smoking and betel nut chewing. The survival rates of patients with advanced OCSCC have not increased significantly in recent years. While treatments for OCSCC are similar worldwide, survival rates differ by geographical area. The various genetic profiles and individual genetic susceptibility for carcinogens may account for this discrepancy. In some respects, molecular alteration or accumulation affects tumor progression and the clinical outcomes among patients with OCSCC. Clarifying the tumor behavior of oral cancer, with regard to pathological features or molecular aspects, could help clinicians to judge, tailor and adopt more effective therapeutic strategies to treat oral cancer.

Objective: To evaluate the prevalence of human papillomavirus (HPV) and the prognostic significance of epidermal growth factor receptor (EGFR), p53, and p16 among patients with oropharyngeal carcinoma., Design: Retrospective study., Setting: Academic Institute of Otolaryngology, Kaohsiung, Taiwan., Patients: Two hundred seventy-four patients who were diagnosed as having oropharyngeal carcinoma underwent testing for the presence of the HPV genome in the nuclei of their tumor cells from January 1, 1992, through March 31, 2008., Interventions: The HPV genome was detected by performing polymerase chain reaction-based assays and in situ hybridization on tumor tissue from paraffin blocks. Immunohistochemistry staining for p16, p53, and EGFR was also performed., Main Outcome Measures: We used the Fisher exact test to evaluate the correlation between the clinicopathological variables and the presence of HPV in tumor cells. Survival analysis was based on the Kaplan-Meier method., Results: We detected HPV in 45 of the 274 patients (16.4%); of these, HPV-16 and -18 were identified in 42 (93.3%) of the HPV-positive tumors. The HPV-positive oropharyngeal cancers were more likely to occur in females, nonsmoking individuals, and those who did not chew betel quid. The HPV-positive tumors significantly expressed p16 and were inversely associated with EGFR and p53 expression (all, P < .001). In addition, patients with tumor tissue that was positive for HPV (P = .008) and had negative expression of EGFR (P = .01), low expression of p53 (P = .01), and high expression of p16 (P = .04) had a better prognosis., Conclusion: Our results suggest that HPV, EGFR, p53, and p16 are useful biomarkers in predicting the clinical outcomes of oropharyngeal cancer.

Objectives: The conventional method for preventing web formation after anterior glottic web surgery is keel insertion. However, this presents risks of airway compromise and granulation tissue formation, which could necessitate tracheotomy in addition to a secondary procedure for keel removal. We introduce a novel, 1-stage endoscopic laser anterior commissurotomy for preventing anterior glottic web re-formation., Methods: Twenty patients with glottic webs involving the anterior commissure were studied. The lesions were removed by transoral carbon dioxide laser microsurgery. In all patients, the anterior glottic web was vaporized along with the inner perichondrium of the thyroid cartilage over the anterior commissure area, creating a raw vertical break "alley" between the anterior vocal folds that measured between 0.3 and 0.5 cm in width and between 0.8 and 2 cm in length. The preoperative and postoperative vocal folds and voice quality were evaluated by videostrobolaryngoscopy and voice recordings., Results: All 20 patients had anterior glottic webs ranging from 11% to 64% of the length from the anterior commissure to the vocal process. None of the patients developed restenosis at the anterior commissure of a severity similar to that of the initial lesion during follow-up (mean, 13 months; range, 3 to 44 months).All patients except 1 reported satisfaction with their voice improvement. Outcome analysis revealed that partial re-formation of the web was noted in 4 patients., Conclusions: One-stage, endoscopic laser anterior commissurotomy was effective and relatively safe for removing glottic webs, for preventing anterior glottic web re-formation, and for improving vocal fold performance among our patients.

Context: Diabetic peripheral neuropathy predisposes patients to foot ulceration that heals poorly and too often leads to amputation. Large-fiber peripheral neuropathy (LFPN), one common form of diabetic neuropathy, when detected early prompts aggressive measures to prevent progression to foot ulceration and its associated morbidity and mortality., Objective: To systematically review the literature to determine the clinical examination findings predictive of asymptomatic LFPN before foot ulceration develops., Data Sources, Study Selection, and Data Extraction: MEDLINE (January 1966-November 2009) and EMBASE (1980-2009 [week 50]) databases were searched for articles on bedside diagnosis of diabetic peripheral neuropathy. Included studies compared elements of history or physical examination with nerve conduction testing as the reference standard., Data Synthesis: Of 1388 articles, 9 on diagnostic accuracy and 3 on precision met inclusion criteria. The prevalence of diabetic LFPN ranged from 23% to 79%. A score greater than 4 on a symptom questionnaire developed by the Italian Society of Diabetology increases the likelihood of LFPN (likelihood ratio [LR], 4.0; 95% confidence interval [CI], 2.9-5.6; negative LR, 0.19; 95% CI, 0.10-0.38). The most useful examination findings were vibration perception with a 128-Hz tuning fork (LR range, 16-35) and pressure sensation with a 5.07 Semmes-Weinstein monofilament (LR range, 11-16). Normal results on vibration testing (LR range, 0.33-0.51) or monofilament (LR range, 0.09-0.54) make LFPN less likely. Combinations of signs did not perform better than these 2 individual findings., Conclusions: Physical examination is most useful in evaluating for LFPN in patients with diabetes. Abnormal results on monofilament testing and vibratory perception (alone or in combination with the appearance of the feet, ulceration, and ankle reflexes) are the most helpful signs.

Background and Objectives: Endoscopic sinus surgery (ESS), markedly improved with the introduction of new preoperative imaging techniques, intraoperative visualization tools, and the use of surgical navigation systems. In this retrospective study we evaluated the usefulness of CT-guided endoscopic sinus surgery and studied its advantages over conventional endoscopic sinus surgery., Methods: We retrospectively reviewed the records of 60 randomly chosen patients with chronic rhinosinusitis (CRS) and moderate-to-severe sinonasal polyposis, undergoing endoscopic sinus surgery with surgical navigation (n=30) and without navigation (n=30). Data on the operative note, time of surgery, complications, and recurrence rate were collected and analyzed., Results: Of the 60 patients, 40 (66.7%) were diagnosed with CRS and 20 (33.3%) had allergic fungal sinusitis. Primary surgery was performed in 37 (61.7%) and revision surgery was performed in 23 (38.3%) cases. The computer-aided surgery (CAS) group included 28 (93.3%) patients with extensive disease and 12 (40%) with bone erosions, with intraorbital or extradural extension, while the non-CAS group included 24 (80%) patients with extensive disease and seven (23.3%) with bone erosions, with intraorbital or extradural extension. The average operative time was approximately 13 minutes greater in the navigation group, with significant improvement in the recurrence rate (n=11, 36.7% in the non-CAS group; n=5, 16.7% in the CAS group), and intraoperative complications were fewer in the CAS group (two exposures of orbital fat in the non-CAS group; no complications in the CAS group)., Conclusion: Computer navigational systems appear to serve as a valuable adjunct in preoperative planning and safe intraoperative dissection.

Objective: To determine the frequency of Non Convulsive Status Epilepticus in patients admitted with impaired consciousness., Methods: All EEG's in patients with impaired level of consciousness over four years from 2002- 2006 were reviewed. All EEG's showing continuous epileptiform discharges were included. Findings of all these EEG's were divided into five groups; generalized spikes and wave, generalized sharp and wave, focal spike and wave, focal sharp and wave and periodic lateralized epileptiform discharges., Results: There were 785 EEG's recorded in patients with impaired level of consciousness. Only 12 (1.5%) patients were identified with NCSE on EEG. The commonest EEG findings in our patients with NCSE were: Continuous focal spike and wave seen in 4(33%), Continuous generalized spike and wave 3 (25%), Continuous generalized sharp and wave 3 (25%), Continuous focal sharp and wave 1 (8.3) and Continuous periodic lateralized epileptiform discharges (PLEDs) in 1 (8.3%) patient., Conclusion: NCSE is an important treatable entity which can be easily recognized by doing an EEG.

Objective: To evaluate the routine use of nasal packing after cosmetic and functional nasal surgery., Methods: Prospective collection from 2005 to 2006 at King Abdulaziz University Hospital in Riyadh for 54 patients post-septorhinoplasty with and without submucous diathermy or lateralization of inferior turbinate were evaluated for efficacy and complications of nasal packing which kept for different duration., Results: All 54 patients had septoplasty, 21 with rhinoplasty, 12 with SMD, and 24 patients had inferior turbinate lateralization. Eight cm slim Merocel packed in 67 nasal cavity sides, 8 cm regular Merocel packed in 24 sides and 10 cm regular Merocel packed in 6 sides. Thirty-seven patients had the pack for 6 hours then removed and 17 patients had it for 24 hours. Forty-one patients with no significant discomfort, 13 patients with mild to moderate discomfort., Conclusion: Septorhinoplasty and SMD or lateralization of inferior turbinate can be managed without packing or any other hemostatic measures.

Objective: To report the allergic fungal sinusitis (AFS) in children in Saudi Arabia and to review the experience of King Abdul-Aziz University Hospital in diagnosis and management of AFS in children., Methods: Hospital charts of 45 children reviewed retrospectively. Clinical presentation, radiological and operative findings, management and outcomes studied. Only 25 patients had =or> 4 diagnostic criteria, treated endoscopically between January 2000 and December 2005 and followed at least 2 years in KAUH, Riyadh, Kingdom of Saudi Arabia., Results: Twenty-five patients had at least 4 criteria for AFS. All patients underwent functional endoscopic sinus surgery (FESS) with high recurrence rate, 44%. Twenty-eight percent needed revision surgery even with medical treatment post operatively. Moreover, no other complications were reported in this study. Aspergillus spp is the most common fungal type in our review., Conclusion: Allergic fungal sinusitis in children is underestimated and understudied associated with poor outcome and high recurrence rate because of difficulty in management. Therefore, the most effective approach of AFS management in children is to have a high index of suspicion, adequate preoperative evaluation, medical preparation preoperatively, meticulous surgery, medical management, postoperative including topical and systemic corticosteroids and close clinical follow-up with endoscopically guided debridement.

Biopolymer (polyhydroxyalkanoate, PHA) was extracted and purified from CMG607w bacterial strain isolated from sediment of Layari River outfall to the Arabian Sea. Synthesis of PHA was substrate depended in CMG607w. In the presence of sodium gluconate mcl-PHA was synthesized at the rate of 42% cell dry mass. Under highly enriched conditions, co-production of polysaccharide and blends of poly(3-hydroxybutyrate) (PHB)/PHA was observed. PCR base strategy was used to amplify PHA polymerase genes from chromosomal DNA. Conserved sequences were observed in polymerase C1 and C2. A portion of polymerase C1 and C2 genes of the pha operon was cloned and sequenced. In CMG607w PHA biosynthsis the operon has phaC1 and C2 (polymerase 1 and polymerase 2) genes. pha polymerase C1/C2 genes of CMG607w have 98% homology to Pseudomonas aeruginosa PAO1 (AE004919). Gene sequences were submitted to GenBank under accession numbers EF028075, AY596788 and AY596790.

The weight of doctors' white coats was examined and correlated to the degree of headache and muscle pain in the shoulders. The coat weight of doctors was 1.5 kg (1.1 kg-1.9 kg) (median with 25%-75% in parenthesis). The highest weight was found in the coats of young physicians 2.0 kg (1.7 kg-2.2 kg), which was higher than those of young surgeons. No correlation was seen in the degree of headache and muscle pain in the shoulders. With the long walking distances in hospitals, it would seem that internal medicine is not only an intellectually, but also a physically, demanding job.

Forty-six female patients with pseudoseizures were compared with 50 female patients referred to a psychiatric outpatient clinic. Patients with convulsive pseudoseizures were more likely to be single and childless. An abnormal EEG was more frequently reported in patients with convulsive pseudoseizures, contributing to their earlier diagnosis of epilepsy. The defence mechanisms of patients with convulsive pseudoseizures using the Defense Mechanisms Inventory (DMI) were particularly characterized by higher scores on the 'reversal' scale and lower scores on the 'turning against self' scale. Mood disorders were more common in general psychiatric patients, while one-third of convulsive pseudoseizure patients did not complain of psychiatric symptoms.

Drawing upon the comprehensive population-based Northern Alberta Breast Cancer Registry containing 704 patients with histologically negative axillary lymph nodes who have been followed for 5-16 years, we have undertaken a retrospective case-control study to evaluate the utility of genomic amplification of specific protooncogenes [c-erbB-2 (nee HER-2/neu), c-erbA, c-myc, int-2, and hst-1] as predictive indicators of clinical outcome in node-negative disease. To this end, 115 women with node-negative breast cancer who had recurred at any time up to 16 years posttreatment (cases) were matched pairwise for appropriate clinicopathological variables (size of primary tumor, menopausal state, estrogen receptor status, anniversary year of treatment, and patient age) with a second group of 115 women (controls) selected from a cohort of 502 node-negative patients who had not relapsed during long-term follow-up. Tumor DNA extracted from archival formalin-fixed, paraffin-embedded tissue blocks were analyzed for protooncogene copy number by slot-blot hybridization. Taking a gene copy number of 3 as the cutoff, 27 of the 230 tumor samples examined contained from 3- to 22-fold elevation in c-erbB-2 genomic equivalents. Twenty-one of the 27 tumors amplified for c-erbB-2 were derived from cases and 6 from controls, signifying that 18% of the node-negative patients who had relapsed harbored excessive copies of the protooncogene in their malignant tissue compared to only 5% for the patients who had remained in remission. Accordingly, the occurrence of amplification of c-erbB-2 proved to be a statistically significant predictor of poor prognosis, especially disease-free interval (P = 0.006). Moreover, this genetic alteration appeared to be independent of and to have greater predictive power than most commonly used prognostic factors. Our findings also indicated that as a clinical test, measurement of c-erbB-2 amplification suffers from low sensitivity; however, when greater than 6 gene copies are present, the test has a positive predictive value for recurrence of 70%. Concurrent analysis of tumor DNA blots with probes for the other four protooncogenes examined revealed that their amplification, which others have reported to arise often, especially in node-positive disease, was seldom found even in our high-risk case group (2-3%). In short, our data strongly suggest that amplification of c-erbB-2 may contribute to the pathogenesis of some forms of node-negative breast cancer and thus may serve as a useful genetic marker to identify a subset of high-risk patients.

Retrospective comparison of peripheral haematological values in normal volunteers and patients on anti-epileptic drugs over a one year period demonstrated a significant reduction in total white blood cells, neutrophil, eosinophil, lymphocyte and monocyte counts in patients. The red blood cell count, mean cell volume and mean cell haemoglobin were significantly reduced in the patients. These findings need to be taken into account when new anti-epileptic drugs are assessed by adding on to existing therapy.

The numbers of T, B, and "null" blood lymphocytes were determined in 216 normal healthy persons, whose ages ranged from 20 to 94 years. No change was found in the numbers of T or B lymphocytes with increasing age of the subjects, whereas both the total lymphocytes and the number of "null" lymphocytes decreased just significantly with age (p less than 0.05). The response of blood lymphocytes to phytohaemagglutinin in vitro was measured in 25 subjects aged 20-30 years and compared with the response in 23 subjects aged over 60 years. There was no significant difference in the response of the two groups.

The photostability of reserpine in several parenteral formulations has been studied and the effects of some commonly used stabilizers have been evaluated. The kinetic data indicate that the photodegradation of reserpine follows zero order kinetics in all the formulations at pH = 2 and 3. The stability of reserpine appears to be higher at pH = 3 than at pH = 2. 3-dehydroreserpine, isoreserpine and lumireserpine were identified as the major photodegradation products by TLC, suggesting that the photodegradation in acidic solutions occurs initially via an oxidative mechanism. The combined effect of EDTA and PABA on the photostability of reserpine was more significant than their individual effect. The formulations in which benzyl alcohol was used as vehicle were found to be more stable than those having propylene glycol. Polyethylene glycol was found to be a better surfactant than Tween 80. The degree of stabilization in the presence of surface active agents has been explained in terms of micellar theory and the degree of hydration of micelles. A stable formula for parenteral solution of reserpine has been suggested.