Genetic diversity of Plasmodium vivax clinical isolates from southern Pakistan using pvcsp and pvmsp1 genetic markers

Background: Plasmodium vivax is the prevalent malarial species accounting for 70% of malaria burden in Pakistan; however, there is no baseline data on the circulating genotypes. Studies have shown that polymorphic loci of gene encoding antigens pvcsp and pvmsp1 can be used reliably for conducting molecular epidemiological studies. Therefore, this study aimed to bridge the existing knowledge gap on population structure on P. vivax from Pakistan using these two polymorphic genes. Methods: During the period January 2008 to May 2009, a total of 250 blood samples were collected from patients tested slide positive for P. vivax, at the Aga Khan University Hospital, Karachi, or its collection units located in Baluchistan and Sindh Province. Nested PCR/RFLP was performed, using pvcsp and pvmsp1 markers to detect the extent of genetic diversity in clinical isolates of P. vivax from southern Pakistan. Results: A total of 227/250 (91%) isolates were included in the analysis while the remainder were excluded due to negative PCR outcome for P.vivax. Pvcsp analysis showed that both VK 210 (85.5%, 194/227) and VK 247 type (14.5%, 33/227) were found to be circulating in P. vivax isolates from southern Pakistan. A total of sixteen and eighty-seven genotypes of pvcsp and pvmsp-1 were detected respectively. Conculsion: This is the first report from southern Pakistan on characterization of P. vivax isolates confirming that extensively diverse pvcsp and pvmsp1 variants are present within this region. Results from this study provide valuable data on genetic diversity of P. vivax that will be helpful for further epidemiological studies

Comparative study between proximal femoral nail and proximal femoral nail antirotation in management of unstable trochanteric fractures

Background: In the management of peri-trochanteric fractures, intramedullary (IM) devices have proven advantage over extramedullary devices. IM devices allow for stable anatomical fixation of more comminuted fractures without shortening the abductor lever arm or changing the proximal femoral anatomy. Between IM devices like proximal femoral nail (PFN) and proximal femoral nail antirotation (PFNA), the helical blade of latter is believed to provide stability, compression and rotational control of the fracture with higher cut out strength. The following study was undertaken in an attempt to compare these two types of Intra-medullary devices.Methods: Between January 2012 and June 2013, 50 patients with unstable intertrochanteric fractures fulfilling inclusion and exclusion criteria, were randomized into 2 groups to undergo CRIF with either standard PFN (n=25) or PFNA (n=25). They were compared in terms of demography, per-operative variables and postoperative parameters including functional evaluation till 1year postoperatively.Results: Background demographic variables, fracture type and pre-injury ambulatory status were comparable between the groups. Operative duration of surgery, amount of blood loss and number of fluoroscopic images were significantly lower in PFNA group as compared to PFN group. Post op complications like infection, non-union, cut out/z-effect, loss of reduction, re-operation and mortality rates didn’t differ significantly between the groups. Post op functional recovery as evaluated by pain, use of walking aids and Harris hip scores were similar in both groups. Conclusions: PFNA significantly reduces the operative time, amount of blood loss and fluoroscopic imaging as compared to PFN. However PFNA offers no significant benefits over PFN in terms of post-operative functional recovery or complications

Prevalence of resistance associated polymorphisms in Plasmodium falciparum field isolates from southern Pakistan

<p>Abstract</p> <p>Background</p> <p>Scarce data are available on <it>Plasmodium falciparum </it>anti-malarial drug resistance in Pakistan. The aim of this study was, therefore, to determine the prevalence of <it>P. falciparum </it>resistance associated polymorphisms in field isolates from southern Pakistan.</p> <p>Methods</p> <p>Blood samples from 244 patients with blood-slide confirmed <it>P. falciparum </it>mono-infections were collected between 2005-2007. Single nucleotide polymorphisms in the <it>P. falciparum </it>chloroquine resistance transporter (<it>pfcrt </it>K76T), multi drug resistance (<it>pfmdr1 </it>N86Y), dihydrofolate reductase (<it>pfdhfr </it>A16V, N51I, C59R, S108N, I164L) and dihydropteroate synthetase (<it>pfdhps </it>A436S, G437A and E540K) genes and <it>pfmdr1 </it>gene copy numbers were determined using PCR based methods.</p> <p>Results</p> <p>The prevalence of <it>pfcrt </it>76T and <it>pfmdr1 </it>86Y was 93% and 57%, respectively. The prevalence of <it>pfdhfr </it>double mutations 59R + 108N/51R + 108N was 92%. The <it>pfdhfr </it>triple mutation (51I, 59R, 108N) occurred in 3% of samples. The <it>pfdhfr </it>(51I, 59R, 108N) and <it>pfdhps </it>(437G, 540E) quintuple mutation was found in one isolate. <it>Pfdhps </it>437G was observed in 51% and 540E in 1% of the isolates. One isolate had two <it>pfmdr1 </it>copies and carried the <it>pfmdr1 </it>86Y and <it>pfcrt </it>76T alleles.</p> <p>Conclusions</p> <p>The results indicate high prevalence of <it>in vivo </it>resistance to chloroquine, whereas high grade resistance to sulphadoxine-pyrimethamine does not appear to be widespread among <it>P. falciparum </it>in southern Pakistan.</p

Genetic diversity among Plasmodium falciparum field isolates in Pakistan measured with PCR genotyping of the merozoite surface protein 1 and 2

Background:The genetic diversity of Plasmodium falciparum has been extensively studied in various parts of the world. However, limited data are available from Pakistan. This study aimed to establish molecular characterization of P. falciparum field isolates in Pakistan measured with two highly polymorphic genetic markers, i.e. the merozoite surface protein 1 (msp-1) and 2 (msp-2).Methods:Between October 2005 and October 2007, 244 blood samples from Patients with symptomatic blood-slide confirmed P. falciparum mono-infections attending the Aga Khan University Hospital, Karachi, or its collection units located in Sindh and Baluchistan provinces, Pakistan were collected. The genetic diversity of P. falciparum was analysed by length polymorphism following gel electrophoresis of DNA products from nested polymerase chain reactions (PCR) targeting block 2 of msp-1 and block 3 of msp-2, including their respective allelic families KI, MAD 20, RO33, and FC27, 3D7/IC.Results:A total of 238/244 (98%) Patients had a positive PCR outcome in at least one genetic marker, the remaining six were excluded from analysis. A majority of Patients had monoclonal infections. Only 56/231 (24%) and 51/236 (22%) carried multiple P. falciparum genotypes in msp-1 and msp-2, respectively. The estimated total number of genotypes was 25 msp-1 (12 KI, 8 MAD20, 5 RO33) and 33 msp-2 (14 FC27, 19 3D7/IC).Conclusion:This is the first report on molecular characterization of P. falciparum field isolates in Pakistan with regards to multiplicity of infection. The genetic diversity and allelic distribution found in this study is similar to previous reports from India and Southeast Asian countries with low malaria endemicity

The Effect Of Ni And Cu Catalysts On The Growth Of Graphene Under Different Ethanol Flow Rates Using Atmospheric Pressure Chemical Vapor Deposition

Graphene was grown on both nickel (Ni) and copper (Cu) catalysts by atmospheric pressure chemical vapor deposition (APCVD) technique at various ethanol flow rates. Raman spectroscopy and field emission scanning electron microscopy (FESEM) were used to study morphological and structural properties of APCVD grown graphene. The crystallite size, defect intensity, distance between defects and the graphene thickness were estimated based on Raman spectra analysis. For the same growth conditions, Ni catalyst promote the formation of more graphene layers as compare to Cu. This because of the higher carbon solubility in Ni as compared to Cu which leads to different growth mechanisms

Predictors of gastrointestinal lesions on endoscopy in iron deficiency anemia without gastrointestinal symptoms

<p>Abstract</p> <p>Background</p> <p>Iron deficiency anaemia (IDA) due to occult gastrointestinal (GI) blood loss usually remains unnoticed until patient become symptomatic. There is sparse data in IDA patients without gastrointestinal symptoms. This study was designed to find out the frequency and predictors of endoscopic lesions in IDA without gastrointestinal symptoms. Cross-sectional study performed on a convenience sample of consecutive subjects.</p> <p>Methods</p> <p>Ninety five consecutive patients with laboratory based diagnosis of IDA having no gastrointestinal symptoms were interviewed and their clinical and biochemical variables were recorded. All the study patients underwent esophago-gastroduodenoscopy (EGD) and colonoscopy. Endoscopic findings were documented as presence/absence of bleeding related lesion and presence/absence of cause of IDA. Multiple logistic regressions were performed to identify variables significantly related to outcome variables.</p> <p>Results</p> <p>Possible cause of anaemia was found in 71% and bleeding related lesions were found in 53% of patients. Upper gastrointestinal tract lesions were found in 41% of patients with bleeding related lesions. On multivariable logistic regression; advancing age, low mean corpuscular volume (MCV ≤ 60 fl), and positive fecal occult blood test were predictive factors for bleeding related GI lesions and cause of IDA</p> <p>Conclusion</p> <p>Clinical and Biochemical markers can predict gastrointestinal lesions on endoscopy in IDA patients without gastrointestinal symptoms. High proportion of upper gastrointestinal involvement warrants EGD as initial endoscopic procedure however, this needs validation by further studies.</p

Diffuse duodenal nodular lymphoid hyperplasia: a large cohort of patients etiologically related to Helicobacter pylori infection

Abstract Background Nodular lymphoid hyperplasia of gastrointestinal tract is a rare disorder, often associated with immunodeficiency syndromes. There are no published reports of its association with Helicobacter pylori infection. Methods From March 2005 till February 2010, we prospectively followed all patients with diffuse duodenal nodular lymphoid hyperplasia (DDNLH). Patients underwent esophagogastroduodenoscopy with targeted biopsies, colonoscopy, and small bowel video capsule endoscopy. Duodenal nodular lesions were graded from 0 to 4 based on their size and density. Patients were screened for celiac sprue (IgA endomysial antibody), immunoglobulin abnormalities (immunoglobulin levels & serum protein electrophoresis), small intestine bacterial overgrowth (lactulose hydrogen breath test), and Helicobacter pylori infection (rapid urease test, and histological examination of gastric biopsies). Patients infected with Helicobacter pylori received sequential antibiotic therapy and eradication of infection was evaluated by 14C urea breath test. Follow up duodenoscopies with biopsies were performed to ascertain resolution of nodular lesions. Results Forty patients (Males 23, females 17; mean age ± 1SD 35.6 ± 14.6 years) with DDNLH were studied. Patients presented with epigastric pain, vomiting, and weight loss. Esophagogastroduodenoscopy showed diffuse nodular lesions (size varying from 2 to 5 mm or more) of varying grades (mean score ± 1SD 2.70 ± 0.84) involving postbulbar duodenum. Video capsule endoscopies revealed nodular disease exclusively limited to duodenum. None of the patients had immunoglobulin deficiency or small intestine bacterial overgrowth or positive IgA endomysial antibodies. All patients were infected with Helicobacter pylori infection. Sequential antibiotic therapy eradicated Helicobacter pylori infection in 26 patients. Follow up duodenoscopies in these patients showed significant reduction of duodenal nodular lesions score (2.69 ± 0.79 to 1.50 ± 1.10; p Helicobacter pylori infection showed no significant reduction of nodular lesions score (2.71 ± 0.96 to 2.64 ± 1.15; p = 0.58). Nodules partially regressed in score in 2 patients, showed no interval change in 10 patients and progressed in 2 patients. Conclusions We report on a large cohort of patients with DDNLH, etiologically related to Helicobacter pylori infection.</p

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]