Epidemiology of EGFR Mutation in Adenocarcinoma NSCLC Patients in India: A Systematic Review and Meta-Analysis

Studies reporting the Indian prevalence of Epidermal Growth Factor Receptor (EGFR) mutation are mostly single centers with small sample sizes. This systematic review and meta-analysis summarized the available evidence of EGFR mutation epidemiology in Indian patients with adenocarcinoma (ADC) Non-Small Cell Lung Cancer (NSCLC). We conducted a structured literature search in PubMed, and EMBASE databases from January 2004 through October 2019. The primary outcome of interest was prevalence of EGFR mutation by gender, smoking status, and mutation subtype. The review included 34 studies. EGFR mutation prevalence was 39.5% in patients with ADC, and significantly higher in females, non-smokers, and patients with exon 19 deletions. The EGFR mutation frequency in Indian patients with ADC was higher than reported in Caucasians but at a lower range of that reported in East Asians. These findings support the use of EGFR mutation testing to guide choice of treatment.

Endoscopic ultrasound guided biliary drainage in surgically altered anatomy: A comprehensive review of various approaches

Endoscopic retrograde cholangiopancreatography(ERCP)is the preferred modality for drainage of the obstructed biliary tree.In patients with surgically altered anatomy,ERCP using standard techniques may not be feasible.Enteroscope assisted ERCP is usually employed with variable success rate.With advent of endoscopic ultrasound(EUS),biliary drainage procedures in patients with biliary obstruction and surgically altered anatomy is safe and effective.In this narrative review,we discuss role of EUS guided biliary drainage in patients with altered anatomy and the various approaches used in patients with benign and malignant biliary obstruction.

Pancreatic fistula after pancreatectomy:Evolving definitions,preventive strategies and modern management

Pancreatic resection is the treatment of choice for pancreatic malignancy and certain benign pancreatic disorders. However, pancreatic resection is technically a demanding procedure and whereas mortality after a pancreaticoduodenectomy is currently < 3%-5% in experienced high-volume centers, post-operative morbidity is considerable, about 30%-50%. At present, the single most significant cause of morbidity and mortality after pancreatectomy is the development of pancreatic leakage and fistula (PF). The occurrence of a PF increases the length of hospital stay and the cost of treatment, requires additional investigations and procedures, and can result in life-threatening complications. There is no universally accepted definition of PF that would allow standardized reporting and proper comparison of outcomes between different centers. However, early recognition of a PF and prompt institution of appropriate treatment is critical to the prevention of potentially devastating consequences. The present article, reviews the evolution of post resection pancreatic fistula as a concept, and discusses evolving definitions, the current preventive strategies and the management of this problem.

Enhanced myocardial fluorodeoxyglucose uptake following Adriamycin-based therapy: Evidence of early chemotherapeutic cardiotoxicity?

AIM: To analyze changes in myocardial glucose metabolism using fluorodeoxyglucose (FDG)-positron emission tomography (PET) in patients treated with adriamycin and to investigate the clinical significance of these changes.METHODS: Considering that FDG-PET scanning has the ability to show changes in glucose metabolism in the myocardium, we retrospectively analyzed the FDGPET studies of 18 lymphoma patients treated with adriamycin-based chemotherapy in both the preand posttherapy setting. Cardiac contractile parameters such as left ventricular ejection fraction were not available for correlation in all patients due to the short duration and the level of cumulative dose administered in these patients during the time of the follow-up FDG-PET study. The change in myocardial glucose utilization was estimated by change in standard uptake values (SUV) in the myocardium.RESULTS: We observed a significant change in SUVmean values in the myocardium (defined as more than change in cardiac SUVmean between pre-and post-chemotherapy PET) in 1 patients, whereas 6 patients did not show any significant cardiac FDG uptake in both preand post-therapy PET scans. Patients were divided into three groups based on the changes observed in myocardial tracer uptake on the followup 18 F-FDG-PET study. Group A (n = 8): showed an increase in cardiac 18 F-FDG uptake in the post-therapy scan compared to the baseline scan carried out prior to starting adriamycin-based chemotherapy. Group B (n = 6): showed no significant cardiac 18 F-FDG uptake in post-therapy and baseline PET scans, and group C (n = 4): showed a fall in cardiac 18 F-FDG uptake in the posttherapy scan compared to the baseline scan. Mean cumulative adriamycin dose (in mg/m 2 ) received during the time of the follow-up FDG-PET study was 256. 25, 250 and 137.5, respectively.CONCLUSION: Our study shows three different trends in the change in myocardial glucose metabolism in patients undergoing adriamycin-based chemotherapy. A further prospective study with prolonged follow-up of ventricular function is warranted to explore the significance of enhanced FDG uptake as a marker of early identification of adriamycin-induced cardiotoxicity.

Psoas muscle metastasis from cervical carcinoma:Correlation and comparison of diagnostic features on FDG-PET/CT and diffusion-weighted MRI

Psoas muscle metastasis, though rare, is the commonest site of skeletal muscle involvement in cervical carcinoma. The appropriate clinical management of this condition, particularly of the pain related to malignant psoas syndrome, is still evolving and the diagnostic features on conventional morphological imaging modalities are often non specific, with the differential diagnosis lying between sarcoma, hematoma, and abscess. In this report, a comparison of various morphofunctional imaging modalities was made. Fluorodeoxyglucosepositron emission tomography(FDG-PET)/computed tomography(CT) was the first to suspect disease involvement of the psoas muscle, demonstrating intense FDG uptake(compared with the contralateral muscle), while ultrasound showed heterogeneous echotexture, and magnetic resonance imaging(MRI) showed subtle altered signal intensity in the right psoas muscle. Both anatomical imaging modalities and non contrast CT of the PET-CT examination demonstrated a bulky psoas muscle, without any focal abnormality. On diffusionweighted imaging of MRI(DWI-MRI), restricted diffusion of the involved muscle was an important observation. The psoas muscle metastatic involvement was proven histopathologically. Thus, enhanced glucose metabolism and restricted diffusion in the newer noninvasive molecular imaging modalities(e.g., PET/CT and DWI-MRI) could serve as valuable adjunctive parameters in diagnosing this entity in the absence of a focal abnormality in the anatomical modalities. In the treatment response monitoring scenario, FDG-PET/CT demonstrated near complete resolution following administration of 3 cycles of systemic chemotherapy and local external radiotherapy.

Is laparoscopic colorectal cancer surgery equal to open surgery? An evidence based perspective

Laparoscopic colorectal surgery(LCS) is an evolving subject.Recent studies show that LCS can not only offer safe surgery but evidence is growing that this new technique can be superior to classical open procedures.Fewer perioperative complications and faster postoperative recovery are regularly mentioned when studies of LCS are presented.Even though the learning curve of LCS is frequently debated when limitations of laparoscopic surgeries are reviewed,studies show that in experienced hands LCS can be a safe procedure for colorectal cancer treatment.The learning curve however,is associated with high conversion rates and economical aspects such as higher costs and prolonged hospital stay.Nevertheless,laparoscopic colorectal cancer surgery(LCCR) offers several advantages such as less co-morbidity and less postoperative pain in comparison with open procedures.Furthermore,the good exposure of the pelvic cavity by laparoscopy and the magnification of anatomical structures seem to facilitate pelvic dissection laparoscopically.Moreover,recent studies describe no difference in safety and oncological radicalness in LCCR compared to the open total mesorectal excision(TME).The oncological adequacy of LCCR still remains unproven today,because long-term results do not yet exist.To date,only a few studies have described the results of laparoscopic TME combined with preoperative adjuvant treatment for colorectal cancer.The aim of this review is to examine the various areas of development and controversy of LCCR in comparison to the conventional open approach.

Diffusion magnetic resonance imaging: A molecular imaging tool caught between hope, hype and the real world of “personalized oncology”

"Personalized oncology" is a multi-disciplinary science, which requires inputs from various streams for optimal patient management. Humongous progress in the treatment modalities available and the increasing need to provide functional information in addition to the morphological data; has led to leaping progress in the field of imaging. Magnetic resonance imaging has undergone tremendous progress with various newer MR techniques providing vital functional information and is becoming the cornerstone of "radiomics/radiogenomics". Diffusionweighted imaging is one such technique which capitalizes on the tendency of water protons to diffuse randomly in a given system. This technique has revolutionized oncological imaging, by giving vital qualitative and quantitative information regarding tumor biology which helps in detection, characterization and post treatment surveillance of the lesions and challenging the notion that "one size fits all". It has been applied at various sites with different clinical experience. We hereby present a brief review of this novel functional imaging tool, with its application in "personalized oncology".

Artificial intelligence in perioperative management of major gastrointestinal surgeries

Artificial intelligence(AI)demonstrated by machines is based on reinforcement learning and revolves around the usage of algorithms.The purpose of this review was to summarize concepts,the scope,applications,and limitations in major gastrointestinal surgery.This is a narrative review of the available literature on the key capabilities of AI to help anesthesiologists,surgeons,and other physicians to understand and critically evaluate ongoing and new AI applications in perioperative management.AI uses available databases called“big data”to formulate an algorithm.Analysis of other data based on these algorithms can help in early diagnosis,accurate risk assessment,intraoperative management,automated drug delivery,predicting anesthesia and surgical complications and postoperative outcomes and can thus lead to effective perioperative management as well as to reduce the cost of treatment.Perioperative physicians,anesthesiologists,and surgeons are well-positioned to help integrate AI into modern surgical practice.We all need to partner and collaborate with data scientists to collect and analyze data across all phases of perioperative care to provide clinical scenarios and context.Careful implementation and use of AI along with real-time human interpretation will revolutionize perioperative care,and is the way forward in future perioperative management of major surgery.

Metastatic clear cell variant of hepatocellular carcinoma with an occult hepatic primary

Metastatic clear cell carcinomas are commonly seen in the kidney and lung. Clear cell variant of hepatocellular carci- noma is an uncommon tumour. Diagnosis is usually made by correlation of histopathology. tumour markers and im- munohistochemistry, especially HepPar 1. In this unusual case of metastatic clear cell carcinoma presenting as Sister Mary Joseph's nodule, no primary evidence was observed radiologically in the liver, but the level of alfa fetoprotein was markedly elevated. Metastatic clear cell carcinoma of the liver with an occult hepatic primary was diagnosed by immunohistochemical profile of the tumour.

Surgery for gallbladder cancer:The need to generate greater evidence

The outcomes for gallbladder cancer remain largely dismal to this day.Overall,the low incidence of gallbladder cancer around the world coupled with an even lower number of patients amenable to surgery at the time of presentation,has precluded the generation of evidence-based guidelines for the management of this cancer.However,while the incidence of the cancer may be decreasing in some parts of the world,in other countries such as India,Japan and Chile,gallbladder cancer continues to affect a sizeable population of patients.As such,there is a growing need to define what constitutes an adequate surgery for each stage of this cancer,based on sound evidence.This editorial provides a broad overview of the existing problems in the management of gallbladder cancer and appeals for multi-institutional studies aimed at answering some of the pertinent questions on the surgical management of gallbladder cancer.

Extended pancreatic resections and lymphadenectomy:An appraisal of the current evidence

Surgery remains the mainstay of treatment for pancreatic ductal adenocarcinoma and complete removal of the cancer confers a definite survival advantage,especially in early disease.However,the majority of patients do not present with early disease,thus precluding the chance of a cure by standard pancreatoduodenectomy(PD),distal pancreatectomy or total pancreatectomy.For this reason,pancreatic surgeons have attempted to push the limits of resection over the last three decades.The aim of these resections has been to determine whether obtaining a complete resection by extending the limits of conventional resection in patients with advanced disease will yield the results seen with PD alone in early disease.This article revisits the data from such studies in an attempt to determine if the available literature supports the performance of extended resections for pancreatic cancer in terms of improvement of survival.

Cell-type specificity of β-actin expression and its clinicopathological correlation in gastric adenocarcinoma

AIM:To investigate cell type specific distribution ofβ-actin expression in gastric adenocarcinoma and its correlation with clinicopathological parameters.METHODS:β-actin is a housekeeping gene,frequently used as loading control,but,differentially expresses in cancer.In gastric cancer,an overall increased expression ofβ-actin has been reported using tissue disruptive techniques.At present,no histological data is available to indicate its cell type-specific expression and distribution pattern.In the present study,we analyzedβ-actin expression and distribution in paired normal and tumor tissue samples of gastric adenocarcinoma patients using immunohistochemistry(IHC),a tissue non-disruptive technique as well as tissue disruptive techniques like reverse transcriptase-polymerase chain reaction(RT-PCR)and western blotting.Correlation ofβ-actin level with clinicopathological parameters was done using univariate analysis.RESULTS:The results of this study showed significant overexpression,at both mRNA and protein level in tumor tissues as confirmed by RT-PCR(1.47±0.13 vs2.36±0.16;P<0.001)and western blotting(1.92±0.26 vs 2.88±0.32;P<0.01).IHC revealed thatβ-actin expression is majorly distributed between epithelial and inflammatory cells of the tissues.Inflammatory cells showed a significantly higher expression compared to epithelial cells in normal(2.46±0.13 vs 5.92±0.23,P<0.001),as well as,in tumor tissues(2.79±0.24 vs6.71±0.14,P<0.001).Further,comparison of immunostaining between normal and tumor tissues revealed that both epithelial and inflammatory cells overexpressβ-actin in tumor tissues,however,significant difference was observed only in inflammatory cells(5.92±0.23vs 6.71±0.14,P<0.01).Moreover,combined expression in epithelial and inflammatory cells also showed significant increase(4.19±0.15 vs 4.75±0.14,P<0.05)in tumor tissues.In addition,univariate analysis showed a positive correlation ofβ-actin level of inflammatory cells with tumor grade(P<0.05)while epithelial cells exhibited negative correlation(P>0.05).CONCLUSION:In gastric cancer,β-actin showed an overall higher expression predominantly contributed by inflammatory or tumor infiltrating immune cells of the tissue microenvironment and correlates with tumor grade.

Cytomegalovirus infection in the bone marrow transplant patient

Cytomegalovirus(CMV) infection is an important contributor to the morbidity and mortality associated with bone marrow transplantation(BMT). Infection may lead to CMV disease involving multiple organs such as pneumonia, gastroenteritis, retinitis, central nervus system involvement and others. CMV seropositivity is an important risk factor and approximately half of BMT recipients will develop clinically significant infection most commonly in the first 100 d post-transplant. The commonly used tests to diagnose CMV infection in these patients include the pp65 antigenemia test and the CMV DNA polymerase chain reaction(PCR) assay. Because of its greater sensitivity and lesser turnaround time, the CMV PCR is nowadays the preferred test and serves as a main guide for pre-emptive therapy. Methods of CMV prevention include use of blood products from seronegative donors or leukodepleted products. Prophylaxis or pre-emptive therapy strategies for CMV prevention may be used post-transplant with the latter becoming more common. The commonly used antivirals for pre-emptive therapy and CMV disease management include intravenous gancyclovir and foscarnet. The role of intravenous immunoglobulin, although used commonly in CMV pneumonia is not clear.

Natural orifice transluminal endoscopic surgery in pancreatic diseases

Natural orifice transluminal endoscopic surgery (NOTES) is a surgical technique that has received considerable interest in recent years. Although minimal access surgery has increasingly replaced traditional open abdominal surgical approaches for a wide spectrum of indications, in pancreatic diseases its widespread use is limited to few indications because of the challenging and demanding nature of major pancreatic operations. Nonetheless, there have been attempts in animal models as well as in the clinical setting to perform diag- nostic and resectional NOTES for pancreatic diseases. Here, we review and comment upon the available data regarding currently analyzed and performed pancreatic NOTES procedures. Potential indications for NOTES include peritoneoscopy, cyst drainage, and necrosectomy, palliative procedures such as gastroenterostomy, as well as resections such as distal pancreatectomy or enucleation. These procedures have already been shown to be technically feasible in several studies in animal models and a few clinical trials. In conclusion, NOTES is a rapidly developing concept/technique that could potentially become an integral part of the armamentarium dealing with surgical approaches to pancreatic diseases.

Histone deacetylase inhibitor pre-treatment enhances the efficacy of DNA-interacting chemotherapeutic drugs in gastric cancer

BACKGROUND The prognosis of gastric cancer continues to remain poor,and epigenetic drugs like histone deacetylase inhibitors(HDACi)have been envisaged as potential therapeutic agents.Nevertheless,clinical trials are facing issues with toxicity and efficacy against solid tumors,which may be partly due to the lack of patient stratification for effective treatments.To study the need of patient stratification before HDACi treatment,and the efficacy of pre-treatment of HDACi as a chemotherapeutic drug sensitizer.METHODS The expression activity of class 1 HDACs and histone acetylation was examined in human gastric cancer cells and tissues.The potential combinatorial regime of HDACi and chemotherapy drugs was defined on the basis of observed drug binding assays,chromatin remodeling and cell death.RESULTS In the present study,the data suggest that the differential increase in HDAC activity and the expression of class 1 HDACs are associated with hypoacetylation of histone proteins in tumors compared to normal adjacent mucosa tissue samples of gastric cancer.The data highlights for the first time that pretreatment of HDACi results in an increased amount of DNA-bound drugs associated with enhanced histone acetylation,chromatin relaxation and cell cycle arrest.Fraction-affected plots and combination index-based analysis show that pre-HDACi chemo drug combinatorial regimes,including valproic acid with cisplatin or oxaliplatin and trichostatin A with epirubicin,exhibit synergism with maximum cytotoxic potential due to higher cell death at low combined doses in gastric cancer cell lines.CONCLUSION Expression or activity of class 1 HDACs among gastric cancer patients present an effective approach for patient stratification.Furthermore,HDACi therapy in pretreatment regimes is more effective with chemotherapy drugs,and may aid in predicting individual patient prognosis.

Second-line treatment of advanced hepatocellular carcinoma:Time for more individualized treatment options?

Hepatocellular carcinoma(HCC)is the most frequently diagnosed primary tumor of the liver and is usually detected as advanced disease.It is an aggressive disease that often progresses rapidly when it fails to respond to treatment.As such,patients have limited opportunities to try different subsequent-line treatment regimens.In the last 5 years,the number of agents and/or regimens available for the treatment of advanced HCC has significantly increased,which has made treatment choices for this patient population increasingly complex.In the secondline setting,several phase III trials of regorafenib(RESORCE),ramucirumab(REACH/REACH-2),and cabozantinib(CELESTIAL)have demonstrated clinically meaningful survival benefits in patients with the disease.However,the median overall survival of patients with advanced HCC remains unchanged at approximately 12 mo from the start of systemic second-line therapy,with a limited duration of response.Evidence from the REACH/REACH-2 trials demonstrated for the first time that baseline alpha-fetoprotein(AFP)levels can be used as an identification factor to select those who are likely to benefit the most from ramucirumab treatment.Ramucirumab is both well tolerated and efficacious and has a clinically acceptable safety profile.Therefore,it should be considered an option for patients with AFP levels≥400 ng/mL.

Comprehensive Genetic Analysis by Integration of Conventional Karyotyping and Interphase FISH Helps Refinement of Biological Subclasses with Clinical Impact in Chronic Lymphocytic Leukemia

Background: Various genetic technologies have been employed in the identification of genomic complexity and refinement of prognostic classification of clinically heterogeneous disease of chronic lymphocytic leukemia (CLL). Objective: The present study of interphase cytogenetics and conventional karyotyping was undertaken to perform comprehensive analysis of CLL genetics with an approach to refine early prognostication of disease. Material & Methods: Retrospective analysis by fluorescence in situ hybridization (FISH) was carried out on total 671 patients of CLL at diagnosis between 2008 and 2015. Conventional cytogenetics studies were performed in 50 of 671 patients using CPG Oligonucleotide + IL-2 and TPA (12-O-Tetradecanyl Phorbol 13-acetate) for stimulation of lymphocytes cultures. Results: Interphase cytogenetics could detect recurrent abnormalities such as del(13q14), +12, del(17p13), del(11q22), del(6q23) in 71% of cases. The incidence of del(13q) was higher in Rai stage 0, I, II (p = 0.0005);whereas patients with ≥2 aberrations were more common in advance stage III, IV (p = 0.001). Frequency of IgH translocation was 7%. Morphology and immunophenotypic analysis revealed atypical CLL with higher frequency of t(14;19) than t(14;18). Conventional karyotype could detect abnormal karyotype in 97% of cases which displayed targeted FISH abnormalities along with additional non-targeted chromosomal abnormalities. Patients with negative FISH markers showed clonal non-recurrent numerical and structural changes. The complex karyotype was identified in 24% cases which included targeted FISH aberrations as well as non-targeted numerical and structural abnormalities like deletions, and unbalanced translocations. A significant association was observed between complex karyotype and coexistence of ≥2 FISH markers (p = 0.009) and del(11q22) &/or del(17p) (p = 0.03). Conclusion: Our data of interphase FISH with integration of conventional karyotyping revealed genomic complexity that helped identification of biological subclasses with clinical impact at diagnosis. Further, these cytogenetic subclasses along with molecular markers are likely to evolve more refined prognostic groups, which will help design risk-adapted therapies in B-CLL.

Characterization of Genomic Events Other than Ph and Evaluation of Prognostic Influence on Imatinib in Chronic Myeloid Leukemia (CML): A Study on 1449 Patients from India

Background: Analysis of Philadelphia (Ph) chromosome, a hallmark of chronic myeloid leukemia (CML) plays an important role in disease monitoring of the targeted drug Imatinib. Apart from Ph, genomic imbalances such as additional chromosomal abnormalities (ACAs) of major route occur during transformation of the disease and show negative impact on prognosis. Objective: The present study was carried out to investigate frequencies of ACAs, genomic deletions, complex Ph variants and their prognostic influences in a large cohort of newly diagnosed CML-CP (chronic phase) and CML-AP/BP (accelerated/blast phase). Material & Methods: Retrospective, single institutional study on 1367 cases of CML-CP and 82 cases of CML-AP/BP between 2009 and 2015, using conventional cytogenetics along with fluorescence in situ hybridization. Results: Of the 1367 patients in CML-CP, 1041 patients who completed 12 - 18 months of Imatinib therapy showed complete cytogenetic remission (CCyR) rates of 76% and 82% at 12 and 18 months respectively. Imatinib induced 81% and 33% CCyR in CML-AP and CML-BP respectively. Frequencies of ACAs in CML-CP, AP and BP were 2%, 27% and 67% respectively. Patients in chronic and AP/BP phase with ACAs showed resistance to Imatinib (p < 0.0005). The incidence of genomic deletions and complex Ph variants was 21% and 6.3% respectively with no comparable difference of cytogenetic response to Imatinib (p p < 0.210 respectively). In a cohort of 112 patients in CCyR, development of new clonal abnormalities, more frequently trisomy 8 was detected in Ph negative clone. Conclusion: Our data demonstrated that Imatinib as a frontline therapy had significantly improved management of CML. However, ACAs play an important role in resistance to Imatinib, both in chronic and acute phase, which may limit sole ABL targeted therapy.

Case of victims of modern imaging technology:Increased information noise concealing the diagnosis

We present a case of tubercular arthritis who underwent numerous unnecessary investigations what is known as "victims of modern imaging technology" or VOMIT. Today there is an exponential rise in the volume of the medical imaging, part of which is contributed by unnecessary and unjustified indications. We discuss about the untoward effects of the uninhibited and careless use of modern imaging modalities and possible ways to avoid. Skeletal manifestation of the tuberculosis is still common in the endemic countries like India. Although the final diagnosis of the skeletal tuberculosis like tubercular arthritis is made by bacteriological and histological studies, few demographic, clinical and radiological features might help making early diagnosis.

In vivo subsite classification and diagnosis of oral cancers using Raman spectroscopy

Oral cancers suffer from poor disease free survival rates due to delayed diagnosis.Noninvasive,rapid,objective approaches as adjuncts to visual inspection can help in better management of oral cancers.Raman spectrosoopy(RS)has shown potential in identification of oral premalignant and malignant conditions and also in the detection of early cancer changes like cancer field ffects(CFE)at buccal mucosa subsite.Anatomic differences between different oral subsites have also been reported using RS.In this study,anatomical differences between subsites and their possible infuence on healthy vs pathological classification were evaluated on 85 oral cancer and 72 healthy subjects.Spectra were acquired from buccal mucosa,lip and tongue in healthy,contralateral(internal healthy control),premalignant and cancer conditions using fiber optic Raman spec-trometer.Mean spectra indicate predominance of lipids in healthy buccal mucosa,contribution of both lipids and proteins in lip while major dominance of protein in tongue spectra.From healthy to tumor,changes in protein secondary structure,DNA and heme related features were observed.Principal component linear discriminant analysis(PC-LDA)followed by leave one out CrOoss-validation(LOOCV)was used for data analysis.Findin gs indicate buccal mucosa and tongue are distinct entities,while lip misclassifies with both these subsites.Additionally,the diagnostic algorithm for individual subsites gave improved classification efficiencies with respect to the pooled subsites model.However,as the pooled subsites model yielded 98%specifcity and 100%sensitivity,this model may be more useful for preliminary screening applications.Large-scale validation studies are a pre requisite before envisaging future clinical applications.