Chromatographic analysis of Hb S for the diagnosis of various sickle cell disorders in Pakistan

Sickle cell disease remains a relatively obscure theme in research on haemoglobinopathies in Pakistan. Limited data is available regarding its prevalence in the country. The objective of our study was not only to estimate the frequency of different sickle cell diseases but also to provide quantitative estimation of haemoglobin S and other haemoglobin variants using an automated high-performance liquid chromatography (HPLC) system. For this Purpose, we retrospectively evaluated the results of HPLC performed on all Patients with suspected haemoglobinopathies during the years 2005 and 2006. Information derived from various sources was used to identify a particular genotype by analysing each sample containing Hb S with respect to haemoglobin, red cell indices and levels of various associated haemoglobin variants. Analysis of 15,699 samples identified 302 Patients with Hb S (1.92%). The genotypes identified included S beta(0)(46.7%), SS (19.2%), SA (11.6%), S beta(+) (8.6%) and SD (2.3%). Thirty-five cases could not be categorised and were labelled \u27unclassified\u27. Majority of the Patients (62.3%) were below the age of 18 years. Balochistan, which is the largest province based on the area, yielded the highest number of Patients (n=140). In the S beta(0) group, the mean haemoglobin and Hb S were lower in children compared to adults (p value of 0.001 and 0.016, respectively). We conclude that sickle cell disorders are prevalent in Pakistan to a significant extent, being concentrated in certain areas of the country. We present the first report of various haemoglobin S genotypes from our population. It is hoped that it will act as a database to characterise the same for our population

Identification of hemoglobin Q india (alpha 1-64 Asp-His) through ARMS-PCR. First report from Pakistan

Various hemoglobinopathies have been reported from Pakistan excepting the rare ones like hemoglobin Q India. Our Purpose of study was to identify the mutation (alpha 1 64 aspartate to histidine) through amplification restriction mutation system-polymerase chain reaction (ARMS-PCR) in Patients where hemoglobin Q has been detected via high performance liquid chromatography (HPLC) and also to evaluate the cost effectiveness of the two technologies. All Patients irrespective of age and gender who underwent HPLC for identification of their hemoglobin variant during January 1, 2006 to January 30, 2007 were studied. The blood samples with unknown peak at a retention time of 4.7 min were evaluated at the molecular level. Analysis of HPLC tracings of 11,008 subjects over a thirteen-month period identified ten individuals with hemoglobin Q. Male to female ratio was 1:1.5 and their age was variable ranging from 1 to 49 (mean 22.8) years. The mean hemoglobin level was 11.3 g/dl while MCV (fl) and MCH (pg) were 73.0 and 20.8 respectively. HPLC showed an unknown peak of 17.7% which was detected as Hb Q. ARMS based PCR showed Hb Q specific product of 370 bp and also an amplified product of 766 bp as the control fragment in these samples. This is the first ever report that documents the presence of Hb Q India (alpha 64 Asp to His) in Pakistani population. We recommend that HPLC be used as a useful screening tool especially in developing countries where PCR facilities may not be accessible

Antibiotic eluting chitosan glycerophosphate implant in the setting of acute bacterial sinusitis: a rabbit model.

BACKGROUND: Topical therapy offers the potential for treatment of sinonasal disease with minimal systemic side effects. Chitosan glycerophosphate (CGP) is a mucoadhesive polymer that can be used as an antibiotic eluting sinonasal implant in the treatment of sinusitis. The purpose of this study was to assess the potential for CGP as an antibiotic impregnated implant in a rabbit model of acute bacterial sinusitis. METHODS: The Institutional Animal Care and Use Committee approved study of acute bacterial sinusitis in 12 New Zealand white rabbits using either Pseudomonas aeruginosa (n = 6) or Staphylococcus aureus (n = 6). CGP impregnated with 50 mg of either gentamicin or vancomycin was bilaterally implanted in two rabbits in each arm, respectively. The sinuses were irrigated with saline for 4 days and the lavage was collected for colony-forming unit (CFU) determination. Within each group, the CFU log reduction in the lavage was compared with that of rabbits receiving saline alone (n = 2) or a daily 80-microgram/mL gentamicin or vancomycin irrigation (n = 2) and analyzed using a Student\u27s t-test. RESULTS: Within the S. aureus group, the CFU log reduction using CGP + vancomycin (-2.57 +/- 0.21) was greater than vancomycin irrigation (-1.66 +/- 0.5; p = NS) and significantly greater than saline alone (2.46 +/- 0.97; p = 0.018). Within the P. aeruginosa group, the CFU log reduction using the CGP + gentamicin (-4.62 +/- 0.74) was greater than gentamicin irrigation (-4.09 +/- 0.70) and saline alone (-1.90 +/- 0.90); however, the results were not significant. In all rabbits receiving the CGP + antibiotic implant, no viable bacteria were present in the lavage by day 4. CONCLUSION: Placement of a single antibiotic impregnated CGP implant in the setting of an acute Gram-positive or Gram-negative bacterial sinusitis resulted in a greater log reduction of CFU than daily antibiotic irrigation and led to complete sterilization of the lavage within 4 days

Renal Tumors in Young Adults A Single-Center Experience From a Developing Country

Purpose: To determine the pattern and outcome of renal tumors in young adults in a large surgical series in Pakistan. Materials and Methods: We retrospectively analyzed 133 young adults (age: ≥ 16 to ≤ 40 years) with 136 renal tumors, who underwent surgical treatment for suspected renal cancer from 1994 till 2010. The clinical and pathological parameters were determined and their impact on final outcome was analyzed. Results: The mean age of the patients was 33.3 ± 6.2 years. Of 136, 121 (88.9%) renal tumors were malignant and 15 (11%) were benign. Among malignancies, 76 (62.7%) patients had stage I or II tumors, 22 (18.1%) stage III, and 23 (19%) stage IV at surgery. The overall cancer-specific survival for malignant tumors at 1, 5, and 10 years was 97%, 83%, and 83%, whereas the cancer-free survival (CFS) was 80%, 63%, and 37%, respectively. Patients with age ≤ 35 years had 1 and 5-year CFS of 83% and 71%, respectively, as compared with 76% and 49% for patients > 35 years (P = .02; odds ratio = 2.3; P = .03). Regarding tumor size, 1 and 5-year CFS for tumors ≤ 10 cm was 93% and 75%, while tumors > 10 cm showed CFS of 56% and 41%, respectively (P = .0001; odds ratio = 4.2; P = .0001). For stage I tumors, CFS at 1 and 5 years was 98% and 84%; for stage II, 82% and 63%; and for stage III, 62% and 50%, respectively. One-year survival for stage IV was 48% only (P = .0001). Conclusion: A wide heterogeneity of renal tumors is seen in young adults with delayed presentation