Molecular Detection of a Pathogenic Entamoeba among Symptomatic Children in Eastern Kurdistan of Iraq.

Entamoeba histolytica infects the large intestine of humans, causing a spectrum of clinical appearances ranging from asymptomatic colonization to severe intestinal and extra-intestinal disease. The parasite is identical microscopically to commensal nonpathogenic amoeba. To detect the pathogenic Entamoeba and estimate the precise prevalence of the parasite among the symptomatic pediatric population using molecular techniques. 323 fecal samples were collected from symptomatic children admitted to Sulaimani Pediatric Teaching Hospital, Sulaimaniyah Province, Iraq, from June to October 2021. A structured, validated questionnaire was prepared and used to report participants' gender, residency, and drinking water source. Then, stool samples were microscopically examined, and the positive samples were submitted to molecular analysis by amplifying the 18s rRNA gene using nested PCR to differentiate E. histolytica from other nonpathogenic Entamoeba. Finally, gene sequences were done to confirm the species. Microscopically, 58 positive samples represented Entamoeba species infection rate of 18% among symptomatic patients. However, only 18 samples were positive for E. histolytica based on molecular methods, which accounts for 31% of the positive by microscopy and 5.6% among the 323 symptomatic populations. NCBI, available in their database, gives the gene sequence and accession number. Patients' sociodemographic data and water sources were directly related to the infection rate. Classical microscopic examination provides a misleading profile about the prevalence of E. histolytica in an endemic region that might lead to unnecessary treatments and a lack of appropriate management for patients.

Safe motherhood: A hidden reality in Pakistan.

OBJECTIVE: To explore the perceptions and experiences of healthcare professionals, including female health visitors, female health workers, community midwives, and heads of departments of healthcare facilities, regarding the importance of the Safe Motherhood Initiative (SMI), its pillars, and its foundational principles. DESIGN: Qualitative descriptive exploratory study. SETTING: This study was conducted in healthcare centres in 10 districts in Pakistan: six in Gilgit Baltistan, two in Chitral, and two in Sindh. PARTICIPANTS: Healthcare professionals were recruited using a purposive sampling technique. In total, 14 in-depth interviews were conducted. FINDINGS: The following themes emerged through thematic analysis: (1) health and well-being of mothers and newborns; (2) strengthening the SMI pillars; (3) equity of health services; and (4) effective strategies for behaviour modification. Each theme had two categories. KEY CONCLUSIONS: The health and well-being of mothers and newborns are key indicators; as such, interventions should be made to promote their quality of life. Capacity building and refresher training on antenatal care, childbirth, postnatal care, postabortion care, and family planning can be considered effective to enhance the competencies of healthcare professionals. IMPLICATIONS FOR PRACTICE: The findings of this study suggest that healthcare professionals should receive training to enhance their competencies and provide safe care.

Midwives' perspectives on perinatal mental health: A qualitative exploratory study in a maternity setting in Karachi, Pakistan.

Women experience perinatal mental health problems (PMHP), such as anxiety and depression antenatally and/or postnatally. Among Asian countries, Pakistan has the highest prevalence of postnatal depression ranging from 28% to 63%. This study aimed to explore midwives' perceptions, understanding, thoughts, and feelings regarding perinatal mental health (PMH), as well as experiences and roles in identifying and managing perinatal women cases suffering from PMHP. A qualitative, descriptive, exploratory design was used. The study was conducted at Koohi Goth Hospital. Purposive sampling was used to recruit practicing midwives holding diplomas in midwifery and public health, and with at least two years of experience. Ten participants were recruited. Data were collected by conducting in-depth interviews using a semi-structured interview guide, translated into Urdu. Interviews were audio recorded. Ethics approval was attained by the institutional Ethical Review Committee. Data analysis was done manually by using the content analysis steps described by Creswell. From the analysis, three themes emerged: 1) Dearth of PMH competencies; 2) Importance of PMH; and 3) Myths related to PMH. PMHP undermine mothers' and infants outcomes, thus, midwives should be competent in assessing and managing symptoms in perinatal women to reduce suffering.

Perpetuation of gender discrimination in Pakistani society: results from a scoping review and qualitative study conducted in three provinces of Pakistan.

BACKGROUND: Gender discrimination is any unequal treatment of a person based on their sex. Women and girls are most likely to experience the negative impact of gender discrimination. The aim of this study is to assess the factors that influence gender discrimination in Pakistan, and its impact on women's life. METHODS: A mixed method approach was used in the study in which a systematic review was done in phase one to explore the themes on gender discrimination, and qualitative interviews were conducted in phase two to explore the perception of people regarding gender discrimination. The qualitative interviews (in-depth interviews and focus group discussions) were conducted from married men and women, adolescent boys and girls, Healthcare Professionals (HCPs), Lady Health Visitors (LHVs) and Community Midwives (CMWs). The qualitative interviews were analyzed both manually and electronically through QSR NVivo 10. The triangulation of data from the systematic review and qualitative interviews were done to explore the gender discrimination related issues in Pakistan. RESULTS: The six major themes have emerged from the systematic review and qualitative interviews. It includes (1) Status of a woman in the society (2) Gender inequality in health (3) Gender inequality in education (4) Gender inequality in employment (5) Gender biased social norms and cultural practices and (6) Micro and macro level recommendations. In addition, a woman is often viewed as a sexual object and dependent being who lacks self identity unless being married. Furthermore, women are restricted to household and child rearing responsibilities and are often neglected and forced to suppress self-expression. Likewise, men are viewed as dominant figures in lives of women who usually makes all family decisions. They are considered as financial providers and source of protection. Moreover, women face gender discrimination in many aspects of life including education and access to health care. CONCLUSION: Gender discrimination is deeply rooted in the Pakistani society. To prevent gender discrimination, the entire society, especially women should be educated and gendered sensitized to improve the status of women in Pakistan.

Highly sensitive nested polymerase chain reaction to improve the detection of Leishmania species in clinical specimens.

Cutaneous leishmaniasis (CL) is one of the most neglected tropical diseases and an important health problem in many countries. It is an endemic disease in most regions of Iraq, while being non-endemic in the Kurdistan Region. The techniques frequently used for detection of CL are not very sensitive. Therefore, this study aimed to identify a sensitive method for diagnosis of CL in clinical samples. The present study was performed in December 2019 to December 2020 in Kalar General Hospital. Clinical samples were collected from 85 suspected CL cases. Sixty-four (75.29%), 71 (83.53%) and 84 (98.82%) cases were detected as positive for CL by microscopy, PCR, and nested PCR, respectively. Of the 84 nested PCR-confirmed CL patients, 46 (54.8%) were female and 38 (45.2%) were male. The most predominate rate of infection was in the 30-39-year age group (29.76%) and the lowest was in the ≥ 60-year group (3.57%). Forty (47.62%) patients had a single lesion. The statistical analysis showed significant differences between age groups and between the number of lesions. The sensitivities of microscopy, conventional PCR, and nested PCR were 80.77%, 86.6% and 100%, respectively, while all three methods showed 100% specificity. Furthermore, PCR-ITS1 followed by a simple restriction fragment length polymorphism (RFLP) analysis using HaeIII endonuclease indicated that Leishmania major was responsible for all CL infections in the study area.

Psychological and situational factors associated with COVID-19 vaccine intention among postpartum women in Pakistan: a cross-sectional study.

OBJECTIVES: Contributing factors to COVID-19 vaccination intention in low-income and middle-income countries have received little attention. This study examined COVID-19-related anxiety and obsessive thoughts and situational factors associated with Pakistani postpartum women's intention to get COVID-19 vaccination. DESIGN: Cross-sectional study administering a survey by a telephone interview format between 15 July and 10 September 2020. SETTING: Four centres of Aga Khan Hospital for Women and Children-Garden, Kharadar, Karimabad and Hyderabad-in Sindh Province, Pakistan. PARTICIPANTS: Women who were enrolled in our longitudinal Pakistani cohort study were approached (n=1395), and 990 women (71%) participated in the survey, of which 941 women who were in their postpartum period were included in the final analysis. PRIMARY OUTCOME MEASURE AND FACTORS: COVID-19 vaccine intention, sociodemographic and COVID-19-related factors, Coronavirus anxiety, obsession with COVID-19 and work and social adjustment were assessed. Multiple multinomial logistic regression analysis was used to identify factors associated with women's intentions. RESULTS: Most women would accept a COVID-19 vaccine for themselves (66.7%). Only 24.4% of women were undecided about vaccination against COVID-19, and a small number of women rejected the COVID-19 vaccine (8.8%). Women with primary education were less likely to take a COVID-19 vaccine willingly than those with higher education. COVID-19 vaccine uncertainty and refusal were predicted by having no experience of COVID-19 infection, childbirth during the pandemic, having no symptoms of Coronavirus anxiety and obsession with COVID-19. Predictors for women's intention to vaccinate themselves and their children against COVID-19 were similar. CONCLUSION: Understanding the factors shaping women's intention to vaccinate themselves or their children would enable evidence-based strategies by healthcare providers to enhance the uptake of the COVID-19 vaccine and achieve herd immunity against Coronavirus.

Molecular-based assay for genotyping Leishmania spp. from clinically suspected cutaneous leishmaniasis lesions in the Garmian area, Kurdistan Region of Iraq.

Cutaneous leishmaniasis (CL) is highly prevalent in southern Iraq and neighboring countries, but is non-endemic to the Kurdistan Region of Iraq, particularly in the Garmian area. This study aimed to investigate the causative agent of CL at the molecular level by amplifying the small subunit (18S) rRNA and internal transcribed spacer 1 (ITS1) region. The present study was conducted from December 2019 to December 2020 at Kalar General Hospital, Kalar, Kurdistan Region, Iraq. Eighty-five clinical specimens were collected selectively from patients with suspected CL lesions via fine needle aspiration. After parasitic genomic DNA was extracted from the removed fluid, PCR and DNA sequencing targeting the 18S rRNA and ITS1 region were performed for molecular detection and species identification. Additionally, for 14 samples, the target bands of amplified DNA fragments for both 18S rRNA and ITS1 were extracted and sequenced via Sanger method using both the directional primers employed in the PCR. Seventy-one (83.53%) of the 85 suspected patients had CL, based on amplification of 18S rRNA and ITS1 via PCR. The sequence analysis revealed that all samples were Leishmania major. Phylogenetic analysis based on ITS1 was also performed. Our study revealed that our molecular method was an efficient technique for detecting CL and a valuable method for identifying Leishmania species in clinical samples. Sequence analysis indicated that the causative agent of CL in the Garmian area was L. major and the disease was rural in origin.

Vitamin D deficiency as a risk factor for urinary tract infection in women at reproductive age.

Vitamin D deficiency is a pandemic problem and an ever-increasing issue in human nutrition and health. Vitamin D (serum 25-hydroxyvitamin D) deficiency causes many health problems such as autoimmune diseases, Crohn's disease, diabetes, inflammation, asthma, hypertension, and cancer. Vitamin D3 (cholecalciferol) deficiency has been documented as a persistent problem among adults, children, and elderly persons in most of the countries. Our main objective of this study was to determine the hypothesis that the vitamin D deficiency among women can lead to them developing frequent urinary tract infections. Vitamin D has a potential role in immune regulation and it prevents infections especially urinary tract infections (UTI). Therefore it has positive regulatory role in both acute and recurrent infections especially in women of reproductive ages. As women at this age group have specific differences in their urinary tract and the reproductive organ anatomy, make them more prone for micro-organisms' invasion, The present study was carried out to ascertain certain relation between serum 25-hydroxyvitamin D levels and UTI in women while contemplating the significance of knowing the risk factors associated with UTI and also finding ways to avoid serious complications. 75 women with (case group) UTI were differentiated with 35 healthy with no UTI (control group) and 40 women with UTI and their serum 25-hydroxyvitamin D levels were checked in a case control study. The women were between at 17-52 years of age. Using ELISA, Serum 25-hydroxyvitamin D levels were measured. Analysis and comparison of the results were done among the two groups. Vitamin D mean levels in the case group was considerably lower when in comparison with the control group (11.09 ± 7.571 ng/mL vs. 24.08 ± 11.95 ng/mL, P < 0.001).

Residual Refractive Astigmatism following Toric Intraocular Lens Implantation without Consideration of Posterior Corneal Astigmatism during Cataract Surgery with Low Anterior Keratometric Astigmatism upto 2.5 Dioptres.

Purpose: To determine the refractive astigmatism following toric intraocular lens (tIOL) implantation without consideration of posterior keratometric astigmatism with a conventional tIOL calculator for eyes with low keratometric astigmatism (0.75D to 2.5D) and to theoretically compare the outcomes with predicted refractive astigmatism using a calculator with Barrett's formula.Methods: 34 eyes (34 patients) were assessed with Scheimpflug imaging and underwent tIOL implantation employing conventional tIOL calculator. Eyes were grouped on preoperative keratometric astigmatism as against-the-rule (ATR), with-the-rule (WTR), and oblique (OB). The refractive astigmatism was assessed at 1, 3, 6 and 12 months postoperatively and was classified as ATR, WTR, and OB. Theoretical refractive astigmatism calculations were performed for the same eyes using Barrett's formula.Results: Preoperatively keratometric astigamtism was ATR, WTR, and OB in 32%, 53% and 15% of eyes. At 12 months, in ATR, WTR and OB groups, 45.5%, 16.7% and 60% had ATR refractive astigmatism; 16.7%, 0%, and 20% had WTR refractive astigmatism; 55.6%, 54.5% and 20% were emmetropic (no sphere and cylinder) respectively. There was a significant difference between the theoretical predicted postoperative refractive astigmatism using conventional tIOL calculator and Barrett's formula (P < .05). Postoperative refractive astigmatism was not significantly different from the theoretical predicted refractive astigmatism with Barrett's formula but it was significantly higher than that with a conventional tIOL calculator.Conclusions: At 12 months, with a conventional tIOL calculator, postoperative emmetropia is achieved in half, two third and one-fifth of eyes with preoperatively ATR, WTR, and OB keratometric astigamtism respectively. Around 1/4th WTR keratometric astigamtism eyes preoperatively were overcorrected to ATR refractive astigmatism whereas ½ATR remained undercorrected at 12 months Outcomes achieved were dissimilar to predicted outcomes with a conventional tIOL calculator but similar to those with Barrett's formula.

Influence of peripheral corneal relaxing incisions during cataract surgery for corneal astigmatism up to 2.5 dioptres on corneal densitometry.

PURPOSE: To assess the effect of peripheral corneal relaxing incisions (PCRI) for astigmatism between 0.75 and 2.5 dioptres during cataract surgery on corneal densitometry (CD). METHODS: In this prospective, randomised study, 80 eyes (80 patients), received either tIOL or PCRI. Assessment at pre-operative and 1, 3, 6, 12 months post-operative visit included uncorrected (UCDVA) and best-corrected distance visual acuity (BCDVA), spherical equivalent (SEQ) (only post-operatively), mean anterior and posterior keratometric astigmatism (KA) and CD with Schiempflug system. CD was analysed in four concentric radial zones from centre to periphery (Zone 1 to 4) and in 3 layers (anterior, mid-stromal and posterior). RESULTS: Comparing tIOLs vs. PCRIs, there was no significant difference in the UCDVA, BCDVA and SEQ. In PCRI group, anterior KA decreased at 1 month and remained stable thereafter. For zones 1, 2 and 3, CD significantly reduced after 3 months with tIOLs whereas with PCRIs, it reduced 1 month onwards. For zone 4, CD reduced only at 12 months with tIOLs compared to 3 months onwards with PCRIs. In both groups CD was higher in the zone 4 and anterior layer. Significant reduction in CD was found in all three layers of cornea after 3 months in tIOL and after first month in PCRI groups, respectively. CONCLUSIONS: Cataract surgery alone reduces the CD. Reducing keratometric astigmatism with PCRIs shows significant differences in CD from early post-operative period for central and anterior corneal layer.

Medication Adherence Apps: Review and Content Analysis.

BACKGROUND: Medication adherence is an expensive and damaging problem for patients and health care providers. Patients adhere to only 50% of drugs prescribed for chronic diseases in developed nations. Digital health has paved the way for innovative smartphone solutions to tackle this challenge. However, despite numerous apps available claiming to improve adherence, a thorough review of adherence apps has not been carried out to date. OBJECTIVE: The aims of this study were to (1) review medication adherence apps available in app repositories in terms of their evidence base, medical professional involvement in development, and strategies used to facilitate behavior change and improve adherence and (2) provide a system of classification for these apps. METHODS: In April 2015, relevant medication adherence apps were identified by searching the Apple App Store and the Google Play Store using a combination of relevant search terms. Data extracted included app store source, app price, documentation of health care professional (HCP) involvement during app development, and evidence base for each respective app. Free apps were downloaded to explore the strategies used to promote medication adherence. Testing involved a standardized medication regimen of three reminders over a 4-hour period. Nonadherence features designed to enhance user experience were also documented. RESULTS: The app repository search identified a total of 5881 apps. Of these, 805 fulfilled the inclusion criteria initially and were tested. Furthermore, 681 apps were further analyzed for data extraction. Of these, 420 apps were free for testing, 58 were inaccessible and 203 required payment. Of the 420 free apps, 57 apps were developed with HCP involvement and an evidence base was identified in only 4 apps. Of the paid apps, 9 apps had HCP involvement, 1 app had a documented evidence base, and 1 app had both. In addition, 18 inaccessible apps were produced with HCP involvement, whereas 2 apps had a documented evidence base. The 420 free apps were further analyzed to identify strategies used to improve medication adherence. This identified three broad categories of adherence strategies, reminder, behavioral, and educational. A total of 250 apps utilized a single method, 149 apps used two methods, and only 22 apps utilized all three methods. CONCLUSIONS: To our knowledge, this is the first study to systematically review all available medication adherence apps on the two largest app repositories. The results demonstrate a concerning lack of HCP involvement in app development and evidence base of effectiveness. More collaboration is required between relevant stakeholders to ensure development of high quality and relevant adherence apps with well-powered and robust clinical trials investigating the effectiveness of these interventions. A sound evidence base will encourage the adoption of effective adherence apps, and thus improve patient welfare in the process.

Toric Intraocular Lenses Versus Peripheral Corneal Relaxing Incisions for Astigmatism Between 0.75 and 2.5 Diopters During Cataract Surgery.

PURPOSE: To compare the outcomes after toric intraocular lens (tIOL) or peripheral corneal relaxing incisions (PCRI) for keratometric astigmatism (KA) between 0.75 and 2.5 diopters (D) during cataract surgery. DESIGN: Prospective randomized clinical trial. METHODS: Eighty eyes (80 participants) received either tIOL or PCRI and were assessed preoperatively, 1, 3, 6, and 12 months postoperatively. PRIMARY OUTCOME MEASURE: Uncorrected (UCDVA) and best-corrected distance logMAR visual acuity (BCDVA) at 12 months. SECONDARY OUTCOME MEASURES: Uncorrected near visual acuity (UCNVA), manifest refraction, KA and mean keratometry (KM), corneal aberrometry, tIOL rotation, and quality-of-life questionnaire. RESULTS: Comparing tIOLs vs PCRIs, there was no significant difference in the UCDVA, BCDVA, and UCNVA. At 12 months, 61% vs 53% had UDCVA of 20/25 or better, 100% vs 76% gained ≥1 lines, and 59% vs 43% were within ±0.13 D spherical equivalent. In the PCRI group, anterior KA decreased at 1 month and remained stable thereafter; there was a nonsignificant trend toward a flatter posterior KA and steeper posterior KM and the total corneal Z2-2 was low at 1 and 12 months. Over 12 months, there were changes in posterior corneal tilt, coma, and hexafoil in the PCRI group. The mean rotation of the tIOLs at 12 months was 1.8 ± 1.4 degrees. tIOL patients were happier and were glad to use the nonprescription sunglasses. CONCLUSION: There was no difference in visual acuity, although more tIOL patients gained ≥1 line and were within ±0.13 D. After PCRIs, the anterior KA decreased in the early postoperative period and remained stable thereafter and posterior corneal aberrations changed constantly over 12 months.

A novel mutation in GP1BA gene leads to mono-allelic Bernard Soulier syndrome form of macrothrombocytopenia.

Inherited macrothrombocytopenia is one of the subgroup of inherited thrombocytopenias with variable bleeding tendencies presenting with low platelet count and giant platelets and different gene mutations are involved in its molecular pathophysiology and affect various cell functions. Herein, we describe a family with an isolated giant platelet disorder with variable bleeding diathesis with autosomal mode of inheritance.

Bengal macrothrombocytopenia is not totally an innocuous condition.

Inherited macrothrombocytopenia is a subgroup of thrombocytopenias, and is characterised by the presence of giant platelets and decreased platelet count with variable bleeding manifestations. Bengal macrothrombocytopenia is a newly described entity, previously called asymptomatic constitutional macrothrombocytopenia (ACMT), presented with variable bleeding tendencies; with mild to severe thrombocytopenia and macro-platelets in their peripheral blood smear and it is not totally an innocuous condition as described previously.

Differential expression of genes involved in Bengal macrothrombocytopenia (BMTCP).

Bengal macrothrombocytopenia (BMTCP) is a giant platelet disorder with mild to moderate thrombocytopenia, clinically characterized by mild bleeding symptoms to totally asymptomatic condition. The pathophysiological mechanism of this condition is not fully understood yet. In the present study, 5 subjects (P1-P5) with BMTCP whose platelet counts ranged between 36140X10(9)/l and mean platelet volume (MPV)13.5-16.1fl were analyzed for differential gene expression of platelets by suppressive subtractive hybridization (SSH) technique. Four genes i.e. myotubularin related protein 9 (MTMR9), iron responsive element binding protein 2 (IREB2), alpha tubulin(TUBA) and tyrosine kinase ligand (TKL) were found to be differentially expressed in patient platelets as compared to that of normal healthy controls which was further confirmed by quantitative RT PCR analysis. The study highlights a multi-factorial etiology for BMTCP which is widely prevalent in the northeastern region of the Indian subcontinent.

The Epidemiology of FVIII Inhibitors in Indian Haemophilia A Patients.

A serious complication of replacement therapy in patients with bleeding disorders is the development of 'inhibitors', particularly FVIII inhibitors in haemophilia A patients. This leads to an increase in the management cost, morbidity and mortality, especially post-operatively. The mechanism of FVIII inhibitor development is quite complex and it is difficult to predict inhibitor development, but a prompt and accurate diagnosis is critical as early therapy can save lives. The aim of this study was to screen patients with bleeding disorders in India for inhibitors, and to analyse and compare the prevalence of inhibitors in different regions in India. Patient details were recorded and blood samples were collected in sodium citrate vacutainers from 1,505 patients with bleeding disorders, in different cities in India. Coagulation and inhibitor screening assays were performed, followed by the Bethesda assay in inhibitor positive samples to quantify the FVIII inhibitor titre. Out of the 1,505 samples analysed, 1,285 were Haemophilia A patients, out of which 78 (6.07 %) were positive for 'FVIII Inhibitors'. The highest incidence of FVIII Inhibitors was seen in South India (13.04 %). The highest incidence of 20.99 % was observed in Chennai, followed by Hyderabad (13.33 %), Jammu (9.90 %) and Guwahati (8.51 %), respectively, with respect to the samples analysed. The other regions showed an inhibitor incidence <8 %. The incidence of inhibitors in haemophilia A patients is different in different regions of India; this may be due to the intensity of treatment, type of product or the genetic characteristics of these patients.

Single cell analysis exposes intratumor heterogeneity and suggests that FLT3-ITD is a late event in leukemogenesis.

FMS-like tyrosine kinase 3 receptor-internal tandem duplication (FLT3-ITD) commonly occurs in acute myeloid leukemia and is considered rare in acute lymphocytic leukemia. Acute leukemia has poor prognosis, mainly due to relapse. Standard FLT3-ITD diagnostic techniques are based on genomic polymerase chain reaction and have recently incorporated GeneScan (Applied Biosystems, Foster City, CA) to identify variations of the FLT3 gene. As this is an average-based assay utilized in a heterogeneous leukemic cell population, we hypothesized that cells of acute leukemia, considered FLT3-ITD-negative by standard methods, could possess a fraction of FLT3-ITD-positive cells. The present study employed single cell mutation analysis to evaluate the FLT3-ITD status in newly diagnosed acute myeloid leukemia (n = 5) and acute lymphocytic leukemia (n = 3) patients. A total of 541 single leukemic cells and 36 mononuclear cells from healthy volunteers were analyzed. Seven patients, considered FLT3-ITD-negative according to bulk DNA analysis, appeared to possess a small fraction of FLT3-ITD-positive cells based on single cell analysis. Moreover, this approach revealed the heterogeneity of the tumor as evident by different FLT3-ITD mutations present in the same patient. The presence of a minor clone carrying FLT3-ITD in almost all patients tested provides evidence that this lesion is a common late event in leukemogenesis. Additionally, 3 relapsed patients demonstrated loss of heterozygosity of the normal allele, affecting 25%-100% of the cells found to be FLT3-ITD-positive. Though further clinical testing is warranted, these findings may have implications on the prognostic significance of FLT3-ITD and the use of targeted therapy.

Novel genetic abnormalities in Bernard-Soulier syndrome in India.

Bernard-Soulier syndrome (BSS) is a severe inherited bleeding disorder due to defects in GPIb/IX/V, a platelet receptor that normally functions as a platelet membrane receptor for von Willebrand factor, thrombin and factor XI. BSS results from mutations in GP1BA, GP1BB or GP9 genes. In 15 patients with Bernard-Soulier syndrome from Western India, we amplified the entire coding sequences of GP1BA, GP1BB and GP9 genes and directly sequenced them. Twelve homozygous changes have been identified, out of which ten were novel mutations. These included eight frameshift mutations, i.e. p.Asp79GlufsX2, p.Phe314PhefsX37, p.Pro93ProfsX59, p.Asp89GlufsX63, p.Glu489AsnfsX64, p.Phe355PhefsX4, p.Leu479PhefsX19 and p.Leu531ArgfsX22, one missense mutation (p.Val262Gly) in GPIBA and one nonsense mutation (p.Tyr95X) in GP9. The two known changes include one missense mutation (p.Cys24Arg) in GP9 and one nonsense change (p.Trp46X) in GPIBB. A wide heterogeneity in the nature of mutations has been observed in Indian BSS patients in the present study. Identification of mutations in this rare platelet function disorder would pave way for genetic diagnosis in affected families in India, where consanguineous marriages are very common.

Molecular pathology of Bernard-Soulier syndrome in Indian patients.

Bernard-Soulier syndrome (BSS) is an extremely rare form of platelet adhesion disorder caused by genetic changes occurring in the glycoprotein genes GPIbα, GPIbß and GPIX with autosomal recessive inheritance pattern. Eight patients from seven unrelated families from Western India were included in this study. Diagnosis of BSS was based on low platelet count, presence of giant platelets in peripheral smear, normal screening coagulation tests, reduced or absence of platelet response to ristocetin in aggregometry studies and reduced or lack of expression of GPIb/IX/V complex on platelet surface in flow cytometry studies. Genomic DNA was screened for mutations in GPIbα, GPIbß and GPIX using direct sequencing method. Six disease causing mutations were detected, out of which four were novel changes including one nonsense (p.Cys32X) in two patients and two missense (p.Tyr95Asp and p.Cys135Tyr) changes in GPIX gene, one insertion mutation (p.Met338fsX13) in GPIbα gene resulting in a frameshift change and two reported mutations, i.e. one insertion mutation in GPIbα gene (p.Val485fsX13) resulting in a new termination codon and one missense change (p.Cys24Arg) in two patients in GPIX gene. The molecular basis of BSS patients presented here shows the heterogeneity of this disorder in Indian patients besides providing a basis for genetic diagnosis of affected families.