scholarly journals Risk Stratification of Patients with Gastrointestinal Stromal Tumours by Histopathological and Immunuhistochemical Analysis

Author(s):  
Prathima Shivaji Rao ◽  
Manikya Manjunath

Introduction: Gastrointestinal Stromal Tumours (GISTs) are the most common mesenchymal tumours of the alimentary canal. Interstitial cells of Cajal are believed to be the cell of origin of GIST which regulates gastrointestinal peristalsis. GISTs encompass a clinicopathologically distinctive, but heterogenous group of neoplasms with reference to their origin, cellular differentiation and prognosis. Aim: The aim of this study was to analyse the histopathological and immunohistochemical characteristics of GIST. Materials and Methods: A hospital-based cross-sectional study of 108 cases of GIST were studied over a period of three years from January, 2017 to December, 2019 in Department of Pathology, Vydehi Institute of Medical Sciences and Research Institute (VIMS&RC), Bangalore. Patient’s demographic data like age and gender were collected. Tumour characteristics like site, size and number of lesions were analysed. Histopathological and immunohistochemistry for Desmin, CD117 and DOG-1 were analysed for significant association with risk groups. Risk stratification according to the Fletcher's risk classification, the cases were divided was divided into very low, low, intermediate and high risk groups. The master chart of collected data was prepared in Excel sheet. Chi-square test with Yates correction was used to calculate p-value to ascertain statistical significance. The master chart of collected data was prepared in Excel sheet. Descriptive statistical analysis was presented in the form of tables, figures and diagrams wherever necessary using Epi info 07 statistical software package. Results: The mean age of the patients was 55.6±14.4 years. Males (55.6%/60 cases) were commonly affected than females (44.4%/48 cases). Small intestine was the commonest location (44.4%/48 cases). Tumour size ranged from 1.5 cm to 25 cm with a mean of 9.1 cm. Microscopic findings revealed that spindle cell type was the most common (86.1%/93 cases). Majority of the cases belonged to high risk category. Multifocality, necrosis, tumour size and >5 mitosis/50 High Power Field (HPF) showed significant association with high risk category tumours (p-value 0.033, 0.016, 0.004 and <0.001, respectively). Immunohistochemical staining showed positivity for CD 117 (83.3%/90 cases), DOG-1 (88.9%/96 cases) and 108 cases/100% negativity for desmin. Conclusion: This study found significant association of high risk groups with tumour size, multifocality, necrosis and mitosis >5/50 HPF. Risk categorisation of GIST remains important. A combination of CD117 and DOG-1 will be more useful in diagnosing GIST rather than using them alone.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 6098-6098
Author(s):  
Winston Wong ◽  
Joseph Cooper ◽  
Steve Richardson ◽  
Bruce A. Feinberg

6098 Background: CareFirst BlueCross BlueShield (CFBCBS) insurance network partnered with Cardinal Health Specialty Solutions (CHSS) to develop a cancer care pathway for network physicians in 2008. The program included a recommendation for molecular diagnostic testing with the Oncotype DX assay for pts with early-stage estrogen receptor-positive breast cancer. Based on NCCN guidelines, the pathway suggested adjuvant chemotherapy for all pts with Oncotype DX Recurrence Scores (RS) in the high-risk category. We aimed to determine the RS risk distribution among pts who received Oncotype DX testing and assess the patterns of care that followed. Methods: Using data from CFBCBS, CHSS proprietary claims software, and Genomic Health, we retrospectively identified a cohort of women with breast cancer who were treated on the CFBCBS clinical care pathways program from 8/2008 to 6/2011 and received Oncotype DX testing. We determined the number of pts with a RS value in the low- (RS <18), intermediate- (RS 18-30), and high-risk (RS ≥31) groups along with the number of pts who subsequently received chemotherapy in each category. Results: Of 1174 women who received Oncotype DX testing, 53% of pts were in the low-, 35% in intermediate-, and 12% in the high-risk groups. Five percent of low-, 41% of intermediate-, and 74% percent of pts in the high-risk category were treated with chemotherapy. Twenty-six percent of pts in the high-risk group did not receive chemotherapy. Conclusions: The proportionate use of chemotherapy in the low and intermediate risk groups was as expected based on adjuvant chemotherapy guidelines; however, the underuse of chemotherapy in 26% of high-risk pts was an unexpected finding. Further study is needed to determine: (1) why physicians avoided chemotherapy in 26% of high-risk pts; (2) the overall number of appropriate pts who underwent Oncotype DX testing; and, (3) the tumor characteristics that may have driven the underutilization of chemotherapy in the high-risk population.


2021 ◽  
Vol 108 (Supplement_6) ◽  
Author(s):  
P Holden ◽  
G Wilson ◽  
M Daniel ◽  
R Srivastava

Abstract Aim Tonsillectomy represents 17% of the elective workload in ENT and post-tonsillectomy haemorrhage is the most significant complication of this procedure. Accordingly, the GIRFT (Getting It Right First Time) report for ENT surgery focusses on the prevention of post-tonsillectomy bleeding. However, there is little guidance on the management of post-tonsillectomy haemorrhage. A local guideline for the management of post-tonsillectomy haemorrhage was introduced in 2020 based on expert consensus. This audit examines the management of patients readmitted with post-tonsillectomy haemorrhage in 2019 and compares this to the management suggested in the new guideline. Method Patients readmitted with post-tonsillectomy haemorrhage within 30 days of a tonsillectomy performed in 2019 were identified. These were retrospectively stratified into risk categories according to both patient and clinical factors. Management was audited against the new guideline including both the initial patient assessment and the treatment suggested for their respective risk category. Results Fifteen patients were identified and stratified into low, medium and high-risk categories. All patients in the “low risk” category were successfully treated conservatively. One patient from the “medium risk” category had a further bleed as an inpatient during the proposed period of observation in the new guideline and was thereafter treated as “high risk”. Within the “high risk” category two patients required return to theatre for arrest of post-tonsillectomy haemorrhage. Conclusions These results show that the risk stratification proposed in these guidelines may be useful in the management of post-tonsillectomy haemorrhage. Amendments to the guideline and a re-audit are in progress.


2021 ◽  
Vol 50 (Supplement_1) ◽  
Author(s):  
Naveen Prashar ◽  
Rahuldeep Singh ◽  
Ritin Mohindra ◽  
Vikas Suri ◽  
Ashish Bhalla ◽  
...  

Abstract Background WHO has declared the COVID-19 as Pandemic on 11th March, 2020. It is important to break the chain of transmission by quarantining the persons with high-risk exposure. Understanding the reasons for quarantine will help in reducing the exposures and thus reducing the chances of quarantine. Methods A validated risk assessment tool based on National Centre for Disease Control guidelines was used for the risk assessment of HCWs. The forms of HCWs who underwent risk assessment between April-November, 2020 were analyzed for reasons of quarantine. The positivity rates among high-risk and low-risk groups were compared. Results Out of 1414 HCWs who were assessed, 345 were categorized as high-risk exposure and were quarantined. The most common reasons for quarantine were performance of aerosol generating procedure without recommended personal protection equipment (PPE) (34%), exposure to COVID-19 positive patient without mask for more than 20 minutes at the distance less than 1 m (30%) and having food/tea together (27%). The positivity rate was 8.4% among high-risk and 1.9% among low-risk exposure group (p-value: &lt;0.001). The positivity among low risk category was more in the second half (19/466; 4.1%) as compared to first half (1/603; 0.2%) of the study period. This might be due to exposure from non-hospital sources as second half coincides with first wave of the pandemic. Conclusion Not using recommended PPE and having tea/food breaks together were the most common reasons for quarantine. Key messages Strict enforcement of recommended PPE and scattered tea and food breaks can reduce high-risk exposures.


Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 104-104
Author(s):  
Paola Guglielmelli ◽  
Terra L. Lasho ◽  
Flavia Biamonte ◽  
Johannah Score ◽  
Carmela Mannarelli ◽  
...  

Abstract Background The median survival (OS) of patients with primary myelofibrosis (PMF; 6.5y) is significantly shortened compared to reference population (Cervantes et al, JCO 2012; 30:2981). OS is predicted by the four risk categories of IPSS, Dynamic-IPSS and DIPSS-plus score, nevertheless pts heterogeneity persists within these categories, necessitating improved risk stratification. We recently reported that pts harboring mutations in any one of the prognostically relevant ASXL1, EZH2, IDH1/2 and SRSF2 genes constitute a IPSS-and DIPPS-plus independent Molecular High Risk category (MHR+) characterized by significant reduction of OS and leukemia free survival (LFS) (Vannucchi et al, Leukemia 2013). The aim of this work was to analyze the impact of the number of prognostically relevant mutated genes on OS and LFS in PMF. Patients and methods Two independent cohorts are included, a “test cohort” from Europe, analyzed at diagnosis, and a “validation cohort” from Mayo Clinic, analyzed at any time after diagnosis. Mutation analysis was performed in DNA from whole blood or granulocytes using RTQ-PCR, HRM and direct sequencing; all mutations were confirmed at least twice. The prognostic value of the molecular variables with regard to OS and LFS was analyzed by Cox regression. Test cohort It included 490 pts (median age 61y; males = 301) risk stratified by IPSS into high (n=74, 15%), intermediate-2 (n=93, 19% ), intermediate-1 (n=147, 30%) and low (n=176, 36%). The median follow-up was 3.63y (95% CI, 0.06-28.33); 161 pts died (33.0%), of whom 76 (15.6%) had progressed to acute leukemia after a median of 3.4y (0.04-28.3) from diagnosis. One hundred forty-six pts (29.8%) presented at least one of the four aforementioned mutated genes and were classified as MHR+. The OS of MHR+ pts was significantly reduced compared to patients with no mutations (n=344): 80.7 vs 148.9 mo (HR 2.2, CI95% 1.6-3.03). One hundred twelve (22.8%) pts had 1 mutation and 34 (6.9%) had 2 or more mutations. In univariate analysis presence of 2 or more mutated genes was significantly more detrimental for OS (29.5mo; HR 4.12, IC95% 2.6-6.4) than having 1 mutated gene (84.2mo; HR 1.8, IC95% 1.2-2.5) compared with having no mutations (148.9mo). Multivariate analysis results adjusted for IPSS indicated that having two or more mutations was an independent prognostic factor for OS (HR 2.9, 95% CI 1.8-4.5). Notably, the prognostic relevance of harboring two or more mutations involved both lower (low and intermediate-1; HR 1.87, 95% CI 1.3-2.6) and higher (intermediate-2 and high; HR 1.6, 95% CI 1.2-2.1) categories of IPSS. Also LFS resulted significantly shorter (129mo; HR 3.1, 95%CI 1.9-4.8) in MHR+ pts compared with pts with no adverse mutations (323mo). Having 2 or more mutations correlated with greater reduction of LFS (79.6mo; HR 7.02, CI95% 3.9-12.6) than having one mutation only (133.9mo; HR 2.2, IC95% 1.3-3.7) as compared with no mutations (323.2mo). Multivariate analysis showed that IPSS high risk category (HR 4.5; 95% CI 2.3-8.8) and two or more mutated genes (HR 5.3; 95% CI 2.9-9.8) predicted independently for a significant reduction in LFS. The negative impact on LFS of having 2 or more mutated genes compared to one or no mutations was maintained in the lower (HR 6.4, 95% CI 2.6-15.2) and higher (HR 5.5, 95% CI 2.3-12.8) risk categories. Validation cohort Validation cohort included 262 patients (median age 64 years; males = 167) risk stratified by DIPSS-plus into high (n=89, 34%), intermediate-2 (n=92, 35%), intermediate-1 (n=46, 18%) and low (n=34, 13%). One hundred forty-six pts (56%) displayed none of the four aforementioned mutations, 93 (36%) harbored one mutation whereas 23 (9%) harbored two or more mutations. In univariate analysis, having two or more mutations (HR 3.7; 95% CI 2.2-6.1) was significantly more detrimental for OS than having no mutations, which is more favorable than having one mutation (HR 1.9; 95% CI 1.4-2.7). When adjusted for DIPSS-plus, the presence of two or more mutations retained its significance (HR 2.1; 95% CI 1.3-3.6) and outperformed ASXL1 mutation alone in its prognostic relevance Conclusions Overall, these results show that the number of prognostically relevant mutated genes correlate with OS and LFS in pts with PMF, suggesting that screening for these mutations might help to improve risk stratification. Disclosures: No relevant conflicts of interest to declare.


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Heba Alshaker ◽  
Robert Mills ◽  
Ewan Hunter ◽  
Matthew Salter ◽  
Aroul Ramadass ◽  
...  

Abstract Background Current diagnostic blood tests for prostate cancer (PCa) are unreliable for the early stage disease, resulting in numerous unnecessary prostate biopsies in men with benign disease and false reassurance of negative biopsies in men with PCa. Predicting the risk of PCa is pivotal for making an informed decision on treatment options as the 5-year survival rate in the low-risk group is more than 95% and most men would benefit from surveillance rather than active treatment. Three-dimensional genome architecture and chromosome structures undergo early changes during tumourigenesis both in tumour and in circulating cells and can serve as a disease biomarker. Methods In this prospective study we screened whole blood of newly diagnosed, treatment naïve PCa patients (n = 140) and cancer-free controls (n = 96) for the presence of 14,241 chromosomal loops in the loci of 425 genes. Results We have detected specific chromosome conformation changes in the loci of ETS1, MAP3K14, SLC22A3 and CASP2 genes in peripheral blood from PCa patients yielding PCa detection with 80% sensitivity and 80% specificity. Further analysis between PCa risk groups yielded prognostic validation sets consisting of HSD3B2, VEGFC, APAF1, BMP6, ERG, MSR1, MUC1, ACAT1 and DAPK1 genes that achieved 80% sensitivity and 93% specificity stratifying high-risk category 3 vs low risk category 1 and 84% sensitivity and 89% specificity stratifying high risk category 3 vs intermediate risk category 2 disease. Conclusions Our results demonstrate specific chromosome conformations in the blood of PCa patients that allow PCa diagnosis and risk stratification with high sensitivity and specificity.


2021 ◽  
Vol 39 (6_suppl) ◽  
pp. 249-249
Author(s):  
Wei Loong Sherman Yee ◽  
Wai Yee Woo ◽  
Adelene Sim ◽  
Kar Perng Low ◽  
Alice Meng ◽  
...  

249 Background: A 22-gene GC has been proposed to refine risk stratification of localized PCa by conventional NCCN criteria, and this may potentially influence treatment recommendations. Nonetheless, majority of studies looking at the utility of GC were conducted in White and non-White men from Western cohorts. We therefore investigated the association of GC with NCCN risk groups (RG) in an Asian PCa cohort. Additionally, we examined for inter-racial differences in molecular subtyping between Asian and White/non-White PCa. Methods: GC (Decipher Biosciences Inc., CA) was performed on diagnostic biopsies of men who were treated with radiotherapy +/- hormonal therapy at a single institution (N = 75). ISUP Gleason’s grade (GG) and tumor cellularity were reviewed by an expert GU pathologist. RNA was extracted from 2 x 2.0-mm tumor cores using Qiagen AllPrep DNA/RNA FFPE Kit (Qiagen, Germany) and gene expression was performed on Affymetrix Human Exon 1.0 ST Array (ThermoFischer, CA). PAM50 molecular subtyping was derived using the DecipherGRID database. Results: We profiled 80 tumors from 75 patients, comprising of 18 (24.0%), 9 (12.0%), 21 (28.0%), and 19 (25.3%) NCCN low-/favorable intermediate-, unfavorable intermediate-, high- and very high-RG, respectively; of note, 8 (10.7%) patients had regional/metastatic disease at diagnosis. Using the GC, 27 (33.8%), 14 (17.5%) and 39 (48.8%) were classified as low- (<0.45), intermediate- (0.45-0.6) and high-RG, respectively (>0.6). When stratified using a three-tier clinico-genomic (CG) classification system (Spratt et al. 2017), 6 of 21 (28.6%) NCCN-defined high-risk and 4 of 19 (21.1%) very high-risk patients were downgraded to CG-defined intermediate-/low-risk, while 2 of 27 (7.4%) NCCN low-/intermediate-risk patients were in fact upgraded to CG high-risk. Next, we interrogated the PAM50 basal-luminal signature in our cohort. Interestingly, when matched to White (N = 5762) and non-White (N = 155) for NCCN RG, ISUP GG and age, we observed a high proportion of basal subtype (62.7%) in Asians, which contrasted the prevalence observed in White (16.7%) and non-White (15.9%) North American patients (Chi-sq P <0.001). Conclusions: Here, we demonstrated the utility of the 22-gene GC for refining the NCCN risk stratification in a largest Asian PCa dataset to-date. An unexpectedly high proportion of PAM50 basal-subtype was observed, suggesting race-specific differences of the tumor transcriptome.


2021 ◽  
Author(s):  
Eun Jung Kwon ◽  
Hye Ran Lee ◽  
Ju Ho Lee ◽  
Mihyang Ha ◽  
Yun Hak Kim ◽  
...  

Abstract Background: Human papillomavirus (HPV) is the major cause of cervical cancer (CC) etiology; its contribution to head and neck cancer (HNC) incidence is steadily increasing. As individual patients’ response to the treatment of HPV-associated cancer is variable, there is a pressing need for the identification of biomarkers for risk stratification that can help determine the intensity of treatment. Methods: We have previously reported a novel prognostic and predictive indicator (HPPI) scoring system in HPV-associated cancers regardless of the anatomical locations by analyzing the TCGA and GEO databases. In this study, we comprehensively investigated the association of group-specific expression patterns of common differentially expressed genes (DEGs) between high-risk and low-risk groups in HPV-associated CC and HNC, identifying a molecular biomarkers and pathways for the risk stratification. Results: Among the identified 174 DEGs, expression of the genes associated with extracellular matrix (ECM)-receptor interaction pathway (ITGA5, ITGB1, LAMB1, LAMC1) were increased in high-risk groups in both HPV-associated CC and HNC while expression of the genes associated with the T-cell immunity (CD3D, CD3E, CD8B, LCK, and ZAP70) were decreased vise versa. The individual genes showed statistically significant prognostic impact on HPV-associated cancers but not on HPV-negative cancers. The expression levels of identified genes were similar between HPV-negative and HPV-associated high-risk groups with distinct expression patterns only in HPV-associated low-risk groups. Each group of genes showed negative correlations, and distinct patterns of immune cell infiltration in tumor microenvironments. Conclusion: These results identify molecular biomarkers and pathways for risk stratification in HPV-associated cancers regardless of anatomical locations. The identified targets are selectively working in only HPV-associated cancers, but not in HPV-negative cancers indicating possibility of the selective targets governing HPV-infective tumor microenvironments.


BMJ Open ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. e030922 ◽  
Author(s):  
Narani Sivayoham ◽  
Lesley A Blake ◽  
Shafi E Tharimoopantavida ◽  
Saad Chughtai ◽  
Adil N Hussain ◽  
...  

ObjectiveTo derive and validate a new clinical prediction rule to risk-stratify emergency department (ED) patients admitted with suspected sepsis.DesignRetrospective prognostic study of prospectively collected data.SettingED.ParticipantsPatients aged ≥18 years who met two Systemic Inflammatory Response Syndrome criteria or one Red Flag sepsis criteria on arrival, received intravenous antibiotics for a suspected infection and admitted.Primary outcome measureIn-hospital all-cause mortality.MethodThe data were divided into derivation and validation cohorts. The simplified-Mortality in Severe Sepsis in the ED score and quick-SOFA scores, refractory hypotension and lactate were collectively termed ‘component scores’ and cumulatively termed the ‘Risk-stratification of ED suspected Sepsis (REDS) score’. Each patient in the derivation cohort received a score (0–3) for each component score. The REDS score ranged from 0 to 12. The component scores were subject to univariate and multivariate logistic regression analyses. The receiver operator characteristic (ROC) curves for the REDS and the components scores were constructed and their cut-off points identified. Scores above the cut-off points were deemed high-risk. The area under the ROC (AUROC) curves and sensitivity for mortality of the high-risk category of the REDS score and component scores were compared. The REDS score was internally validated.Results2115 patients of whom 282 (13.3%) died in hospital. Derivation cohort: 1078 patients with 140 deaths (13%). The AUROC curve with 95% CI, cut-off point and sensitivity for mortality (95% CI) of the high-risk category of the REDS score were: derivation: 0.78 (0.75 to 0.80); ≥3; 85.0 (78 to 90.5). Validation: 0.74 (0.71 to 0.76); ≥3; 84.5 (77.5 to 90.0). The AUROC curve and the sensitivity for mortality of the REDS score was better than that of the component scores. Specificity and mortality rates for REDS scores of ≥3, ≥5 and ≥7 were 54.8%, 88.8% and 96.9% and 21.8%, 36.0% and 49.1%, respectively.ConclusionThe REDS score is a simple and objective score to risk-stratify ED patients with suspected sepsis.


Author(s):  
Pinky Karam ◽  
B. Shanthi ◽  
Kalai Selvi

Background: Metabolic syndrome is a group of metabolic abnormalities in which the chance of developing cardiovascular disease, diabetes mellitus, chronic kidney disease are high. Aim: It aims at studying the lipid abnormalities in metabolic syndrome patients. Methods: Total of 100 metabolic syndrome patients were selected for study over a period of 1year. These patients were selected based on the criteria for metabolic syndrome as established by National Cholesterol Education Program (NCEP) adult Treatment Panel III (ATP III). Demographic data were taken and biochemical parameters were estimated by standard guideline. Results: Total cholesterol is significantly higher in very high risk (272.1 ± 8.591) compared to high risk (241.2 ± 3.901) and moderate risk (231.5 ± 4.498). TGL is significantly higher in very high risk (263.9 ± 13.70) compared to high risk (202.1 ± 6.531) and moderate risk (183.7 ± 7.650). HDL is almost same in very high risk (43.09 ± 1.533), high risk (40.44 ± 0.996) and moderate risk (42.53 ± 1.088). LDL is significantly higher in very high risk (177.9 ± 4.255) and high risk (169.4 ± 3.190) compared to moderate risk (155.7 ± 3.098). VLDL is significantly higher in very high risk (52.78 ± 2.739) compared to high risk (40.43 ± 1.306) and moderate risk (36.73 ± 1.530). CHO: HDL is significantly higher in very high risk (6.648 ± 0.366) compared to moderate risk (5.560 ± 0.207). High risk (6.060 ± 0.156) is not significantly different from very high risk and moderate risk. Thus, TC, TGL, LDL, VLDL, and CHO: HDL is significant as p value < 0.05 while HDL did not have any significance as p value > 0.05. Conclusion: In this study, high prevalence of dyslipidaemia is seen. So, timely diagnosis and treatment will help in detecting dyslipidaemia patients in future.


2014 ◽  
Vol 6 (3) ◽  
pp. 49-57 ◽  
Author(s):  
Fiekumo Igbida Buseri ◽  
Charity Ngozi Okonkwo

Background: This study aims at investigating the seroprevalence of HIV infection among status naive pregnant women and probable vertical transmission in Sokoto, Nigeria.Materials and Methods: This cross-sectional study examined 13,026 apparently healthy pregnant women aged between 14 and 45 years and 312 mother-baby pairs in 4 different hospital settings in Sokoto State, North West, Nigeria between March, 2011 and February, 2013. The babies were aged between 8 and 16 months. HIV screening was performed using qualitative rapid tests and ELISA and HIV-DNA polymerase chain reaction (PCR) techniques. Measurement of CD4+ T-lymphocytes was carried out by the BD FACScount System. All seropositive pregnant women were immediately placed on triple antiretroviral therapy (ART) throughout the duration of the pregnancy and beyond.Results: An overall 2.4% prevalence of HIV-1 infection among the pregnant women and 20.5% incident of mother-to-child transmission were found. Of the seropositive pregnant women, 75.0% were full-time house wives, 13.8% and 11.2% were traders and civil servants respectively; of which, 70.2% were within the ages of 14 and 27 years (youthful predominance). Pearson’s χ2analysis did not show any statistically significant difference in the Mean values in the 4 health facilities (χ2 =2.084, df=3, P-value=0.555). Similarly, no significant difference in HIV seropositivity in the demographic data of the pregnant women were observed (P>0.05). Infection was recorded in all age groups but there was no statistical significance between age groups and infection (P = 0.833). Of the 64 seropositive babies, 62 (92.5%) contracted HIV from antiretroviral therapy non-adherence mothers (χ2 =271.457, df=1, P<0.01), OR=1506.6 (95%CI=285.5-7950.4). Conclusion: This study found high prevalence of vertical transmission due to ART non-adherence. Intervention initiatives should, therefore, focus seriously on ART non-adherence. DOI: http://dx.doi.org/10.3126/ajms.v6i3.11530Asian Journal of Medical Sciences Vol.6(3) 2015 49-57  


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