Development of a rapid and sensitive real-time diagnostic assay to detect and quantify Aphanomyces invadans, the causative agent of epizootic ulcerative syndrome.

The oomycete Aphanomyces invadans causes epizootic ulcerative syndrome (EUS), a World Organization for Animal Health (WOAH)-listed disease that has seriously impacted a wide range of fish worldwide. Currently, only three conventional polymerase chain reaction (PCR) assays are recommended for the detection of A. invadans. The robust quantitative PCR (qPCR) assay has recently become more important due to its highly accurate nature and the applicability of qPCR-based environmental DNA (eDNA) detection in the monitoring of pathogens in aquatic environments. Therefore, in this study, we developed a novel TaqMan probe-based qPCR method to sensitively and quantitatively detect A. invadans. The assay limit of detection was determined using 10-fold serial dilutions of linearized A. invadans plasmid. Assay sensitivity was assessed in the presence of interfering substances and compared to three WOAH-listed primers using the mycelia and zoospores of A. invadans with and without fish muscle tissue. The assay specificity was also theoretically and experimentally assessed against other oomycetes, fish muscle tissue, and water samples. The assay's repeatability and reproducibility were determined. In this study, the limit of detection of the developed assay was 7.24 copies of A. invadans genomic DNA per reaction (95% confidence interval (CI): 2.75 to 19.05 copies/reaction). The assay showed the same sensitivity in the presence of other substances. Compared to the WOAH-recommended PCR assays, this assay had 10-times higher sensitivity for all tested samples. There were no cross-reactions with other closely related oomycetes, fish muscle, or water samples, indicating that the assay was highly specific for A. invadans. The repeatability and reproducibility tests showed little variation, ranging from 0.1-0.9% and 0.04-1.1%, respectively, indicating the high consistency, repeatability, and reliability of the developed assay. This highly rapid, sensitive, specific, and consistent EUS qPCR assay would be of importance in transboundary disease management and the monitoring of pathogens in aquatic environments.

Decreased mortality in coronavirus disease 2019 associated mucormycosis with aspirin use: a retrospective cohort study.

OBJECTIVE: Rhino-orbito-cerebral mucormycosis is a rapidly progressive disease with high mortality rates of about 60 per cent. The increasing incidence of rhino-orbito-cerebral mucormycosis in coronavirus disease 2019 patients in India and worldwide has become a matter of concern owing to the case fatality rate. This study explored the use of low dose aspirin in decreasing the mortality rate of coronavirus disease 2019 associated mucormycosis. METHOD: This was a retrospective observational study. Patients suffering from post-coronavirus disease 2019 mucormycosis were included in the study. Each patient was treated with surgical debridement and systemic amphotericin B. Low dose aspirin was added, and mortality rates were compared with the patients who did not receive aspirin. RESULTS: The demographic data and rhino-orbito-cerebral mucormycosis staging between the two groups were not significantly different. There was a statistically significant difference in mortality outcomes between the two groups (p = 0.029) and a 1.77 times higher risk of dying for patients not receiving aspirin. Kaplan-Meier survival indicated that patients receiving aspirin had better survival rates (p = 0.04). CONCLUSION: Low dose aspirin improves survival rates in coronavirus disease 2019 associated mucormycosis.

Mechanisms associated with the rapid decline in sea ice cover around a stranded ship in the Lazarev Sea, Antarctica.

In the satellite data era starting from 1979, the extent of Antarctic sea ice increased moderately for the first 37 years. However, the extent decreased to record low levels from 2016 to 2020, with the drop being greatest in the Weddell and Lazarev Seas of the Southern Ocean. An important question for the scientific fraternity and policymakers is to understand what ocean-atmospheric processes triggered such a rapid decline in sea ice. We employ in-situ, satellite, and atmospheric reanalysis data to examine the causative mechanism of anomalous sea ice variability in the Lazarev Sea at a time of ice growth in the annual cycle (March-April 2019), when a cargo ship was stuck in extensive ice cover and freed following the unusual decline in sea ice. High-resolution Sentinel-1 synthetic aperture radar captured a distinct view of the ship location and track within extensive ice cover of fast sea ice, dense pack ice, and icebergs in the Lazarev Sea on 27 March 2019. Subsequently, the sea ice cover declined and reached the fourth lowest extent in the entire satellite record during April 2019 which was 25.6% lower than the long-term mean value of 2.65 × 106 km2. We show that the anomalous sea ice variability was due to the occurrence of eastward-moving polar cyclones, including a quasi-stationary explosive development that impacted sea ice through extreme changes in ocean-atmospheric conditions. The cyclone-induced dynamic (poleward propagation of ocean waves and ice motion) and thermodynamic (heat and moisture plumes from midlatitudes, ocean mixed layer warming) processes coupled with high tides provided a conducive environment for an exceptional decline in sea ice over the region of ship movement.

Research Capacity for Prevention and Control of Non-communicable Diseases and their Risk Factors in Nepal: Findings of a Needs Assessment Study.

Background Despite a high burden, there are limited trainings in non-communicable disease research in Nepal. Objective We conducted a needs assessment to assess existing research training capacity in academic health institutions of Nepal for the prevention and control of noncommunicable diseases, identify gaps in research, and explore the feasibility of developing research training program in Nepal targeting non-communicable diseases. Method We did mixed-method research and reviewed academic institution curricula and scientific articles authored by Nepali researchers. We conducted 14 Focus Group Discussions with bachelor and masters level students of public health and community medicine; 25 In-depth Interviews with department heads and faculties, and government stakeholders. We surveyed medical and public health students on their research knowledge and skills development. Result Research methodology component was addressed differently across academic programs. One-third (33.7%) of students expressed lack of skills for analysis and interpretation of data. They felt that there is a wide scope and career-interest in non-communicable diseases research in Nepal. However, specific objectives in the curriculum and practical aspects regarding non-communicable diseases were lacking. Most of the non-communicable diseases research in Nepal are prevalence studies. Lack of funding, conflicting priorities with curative services, and inadequate training for advanced research tools were reported as major barriers. Conclusion Nepal must strengthen the whole spectrum of research capacity: epidemiological skills, research management, and fund development. Generation of a critical mass of non-communicable disease researchers must go together with improved funding from the government, non-governmental, and external funding organizations.

Perception of Medical Students Regarding TU-IOM MBBS Curriculum and Teaching Learning Methods in Nepal.

Background The present Bachelor of Medicine and Bachelor of Surgery (MBBS) curriculum under Tribhuvan University - Institute of Medicine (TU-IOM) was last revised twelve-years back. Though the curriculum was built upon internationally approved recommendations on curriculum design, it is ineffectively practiced in most medical schools of Nepal with major focus on didactic teaching-learning. The curriculum, hence, needs effective implementation and revision. Objective To identify the strengths, weaknesses, and areas of improvement in the medical curriculum through student-based feedback and outline the possibility of incorporating newer evidence-based teaching-learning methodologies in Nepal. Method This is a descriptive and cross-sectional study. With appropriate ethical approval, a questionnaire was developed and disseminated virtually to all medical students of Nepal under TU from MBBS fourth year onwards. The questionnaire comprised of Likert and close-ended questions. The data analysis was followed after receiving the filled questionnaire through Google forms. Result A total of 337 respondents participated in the study. The most effectively implemented components out of the SPICES model were Integrated learning (I) and Communitybased learning (C), with 73.89% and 68.84% responses. There were 94.7% (319) students who favored the incorporation of research in the core curriculum. Only 34.2% (115) students found PowerPoint lectures, the most utilized form of teachinglearning in Nepal, as engaging. The respondents (84.6%) showed a high degree of readiness to incorporate newer evidence-based teaching-learning tools such as flipped learning, blended learning, and peer-to-peer learning. Conclusion This study shows that effective interventions must be rethought on various aspects of the curriculum, taking students' feedback on the table while considering curricular revision.

Reassortment and evolutionary dynamics of tilapia lake virus genomic segments.

The tilapia lake virus (TiLV), a highly infectious negative-sense single-stranded segmented RNA virus, has caused several outbreaks worldwide since its first report from Israel in 2014, and continues to pose a major threat to the global tilapia industry. Despite its economic importance, little is known about the underlying mechanisms in the genomic evolution of this highly infectious viral pathogen. Using phylogenomic approaches to the genome sequences of TiLV isolates from various geographic regions, we report on the pervasive role of reassortment, selection, and mutation in TiLV evolution. Our findings provided the evidence of genome-wide reassortment in this newly discovered RNA virus. The rate of non-synonymous (dN) to synonymous (dS) substitutions was less than one (dN/dS = 0.076 to 0.692), indicating that each genomic segment has been subjected to purifying selection. Concurrently, the rate of nucleotide substitution for each genomic segment was in the order of 1-3 × 10-3 nucleotide substitutions per site per year, which is comparable to the rate of other RNA viruses. Collectively, in line with the results of the previous studies, our results demonstrated that reassortment is the dominant force in the evolution and emergence of this highly infectious segmented RNA virus.

Prevalence of methicillin-resistant Staphylococcus aureus in a tertiary hospital in Nepal.

SETTING: Patan Hospital, Lalitpur, Nepal. OBJECTIVES: To describe 1) the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and its antibiotic sensitivity pattern; 2) the demographic and clinical characteristics associated with MRSA infections; and 3) the treatment outcomes of in-patients with MRSA infection among patients with S. aureus infection between January 2018 and December 2020. DESIGN: This was a cross-sectional study using electronic and paper-based hospital records of patients with S. aureus infection. RESULTS: Of the 1,804 patients with S. aureus infection, 1,027 patients (57%, 95% CI 55-59) had MRSA. The MRSA were susceptible to vancomycin (100%), linezolid (96%), doxycycline (96%), chloramphenicol (86%) and cotrimoxazole (70%), and resistant to erythromycin (68%), clindamycin (56%), gentamycin (58%), ciprofloxacin (92%) and ofloxacin (91%). The prevalence of MRSA was higher in 2019, among out-patients, and in respiratory samples, and lower in blood samples. Of the 142 in-patients with MRSA, 93% had a successful clinical outcome (cured/improved). CONCLUSION: More than 50% of patients with S. aureus infection had MRSA that were resistant to commonly available antibiotics. This calls for strengthening surveil-lance and good infection control practices in this hospital.

LIEU: Hôpital de Patan, Lalitpur, Népal. OBJECTIFS: Décrire 1) la prévalence de Staphylococcus aureus résistant à la méticilline (MRSA) et son profil de sensibilité aux antibiotiques ; 2) les caractéristiques démographiques et cliniques associées aux infections à MRSA ; et 3) les résultats thérapeutiques des patients hospitalisés atteints d'infection à MRSA parmi ceux atteints d'infection à S. aureus de janvier 2018 à décembre 2020. MÉTHODE: Il s'agissait d'une étude transversale réalisée en utilisant les dossiers hospitaliers électroniques et papiers des patients atteints d'infection à S. aureus. RÉSULTATS: Sur les 1 804 patients atteints d'infection à S. aureus, 1 027 patients (57%, IC 95% 55-59) avaient un MRSA. Les MRSA étaient susceptibles à la vancomycine (100%), au linézolide (96%), à la doxycycline (96%), au chloramphénicol (86%) et au co-trimoxazole (70%), et résistants à l'érythromycine (68%), la clindamycine (56%), la gentamycine (58%), la ciprofloxacine (92%) et l'ofloxacine (91%). La prévalence des MRSA était plus élevée en 2019, parmi les patients ambulatoires, ainsi que dans les échantillons respiratoires. Elle était plus faible dans les échantillons sanguins. Sur les 142 patients hospitalisés avec MRSA, 93% ont connu un résultat clinique favorable (guérison/amélioration de l'état). CONCLUSION: Plus de 50% des patients atteints d'infection à S. aureus avaient un MRSA résistant aux antibiotiques habituellement disponibles. La surveillance et les pratiques de contrôle des infections doivent donc être renforcées dans cet hôpital.

TRAF6-IRF5 kinetics, TRIF, and biophysical factors drive synergistic innate responses to particle-mediated MPLA-CpG co-presentation.

Innate immune responses to pathogens are driven by co-presentation of multiple pathogen-associated molecular patterns (PAMPs). Combinations of PAMPs can trigger synergistic immune responses, but the underlying molecular mechanisms of synergy are poorly understood. Here, we used synthetic particulate carriers co-loaded with monophosphoryl lipid A (MPLA) and CpG as pathogen-like particles (PLPs) to dissect the signaling pathways responsible for dual adjuvant immune responses. PLP-based co-delivery of MPLA and CpG to GM-CSF-driven mouse bone marrow-derived antigen-presenting cells (BM-APCs) elicited synergistic interferon-ß (IFN-ß) and interleukin-12p70 (IL-12p70) responses, which were strongly influenced by the biophysical properties of PLPs. Mechanistically, we found that MyD88 and interferon regulatory factor 5 (IRF5) were necessary for IFN-ß and IL-12p70 production, while TRIF signaling was required for the synergistic response. Both the kinetics and magnitude of downstream TRAF6 and IRF5 signaling drove the synergy. These results identify the key mechanisms of synergistic Toll-like receptor 4 (TLR4)-TLR9 co-signaling in mouse BM-APCs and underscore the critical role of signaling kinetics and biophysical properties on the integrated response to combination adjuvants.

Needs Assessment of Leadership and Governance in Cardiovascular Health in Nepal.

Background Good governance and leadership are essential to improve healthy life expectancy particularly in low and middle-income countries (LMICs). This study aimed to epitomize the challenges and opportunities for leadership and good governance for the health system to address non-communicable diseases particularly cardiovascular diseases (CVD) in Nepal. Objective The objective of this study was to understand and document CVD programs and policy formulation processes and to identify the government capacity to engage stakeholders for planning and implementation purposes. Method A national-level task force was formed to coordinate and steer the overall need assessment process. A qualitative study design was adopted using "The Health System Assessment Approach". Eighteen indicators under six topical areas in leadership and governance in cardiovascular health were assessed using desk review and key informant interviews. Result Voice and accountability exist in planning for health from the local level. The government has shown a strong willingness and has a strategy to work together with the private and non-government sectors in health however, the coordination has not been effective. There are strong rules in place for regulatory quality, control of corruption, and maintaining financial transparency. The government frequently relies on evidence generated from large-scale surveys for health policy formulation and planning but research in cardiovascular health has been minimum. There is a scarcity of cardiovascular disease-specific protocols. Conclusion Despite plenty of opportunities, much homework is needed to improve leadership and governance in cardiovascular health in Nepal. The government needs to designate a workforce for specific programs to help monitor the enforcement of health sector regulations, allocate enough funding to encourage CVD research, and work towards developing CVD-specific guidelines, protocols, and capacity building. KEY WORDS Cardiovascular diseases, Governance, Leadership, Needs assessment, Nepal.

Transcriptome analysis reveals immune pathways underlying resistance in the common carp Cyprinus carpio against the oomycete Aphanomyces invadans.

Infection with Aphanomyces invadans is a serious fish disease with major global impacts. Despite affecting over 160 fish species, some of the species like the common carp Cyprinus carpio are resistant to A. invadans infection. In the present study, we investigated the transcriptomes of head kidney of common carp experimentally infected with A. invadans. In time course analysis, 5288 genes were found to be differentially expressed (DEGs), of which 731 were involved in 21 immune pathways. The analysis of immune-related DEGs suggested that efficient processing and presentation of A. invadans antigens, enhanced phagocytosis, recognition of pathogen-associated molecular patterns, and increased recruitment of leukocytes to the sites of infection contribute to resistance of common carp against A. invadans. Herein, we provide a systematic understanding of the disease resistance mechanisms in common carp at molecular level as a valuable resource for developing disease management strategies for this devastating fish-pathogenic oomycete.

Perception of Students on Online Assessment: A Descriptive Study among Medical Undergraduates at Basic Sciences.

Background Assessment drives learning. Student assessment cannot be neglected in the long ongoing online learning during COVID-19 pandemic to motivate and assess students' learning. Hence Patan Academy of Health Sciences (PAHS) conducted online assessment to medical undergraduates amidst limitations. Objective To measure the perception of online assessment of students which will provide applicable insights for the further improvement. Method This is a single center, cross-sectional and descriptive study. A Google form containing a semi-structured questionnaire was sent to MBBS students of Basic Science at PAHS who attended online classes and online examination. The responses from close ended questions expressed in percentage and Chi-square test was used to find the association. Open ended questions were analyzed using Braun and Clarke's thematic analysis. Result Of 118 students that responded, 75% passed the online examination. A majority of students (73%) stated that online exam motivated them in learning process and it could be a good alternative during pandemic time. However the most of students (56%) were unsatisfied with the modality of online assessment and reasons that they mentioned were technical problems (89%), inadequate online proctoring (77%), insufficient examination time (58%), lack of orientation to exam, lack of computer skills. Technical problem was worse in rural areas. Conclusion Even though online assessment motivated students' learning during online classes, a large number of students were unsatisfied with assessment modality.

Lutetium therapy-induced carcinoid crisis: A case report and review of literature.

Peptide receptor radionuclide therapy with 177lutetium (Lu)-labeled somatostatin analogs is a promising new tool in the management of patients with inoperable or metastatic neuroendocrine tumors. Some of these patients may present with carcinoid syndrome and it is known that rarely carcinoid crisis can be precipitated by surgical or other interventions in these patients. However, there are anecdotal reports of carcinoid crisis after Lu-labeled peptide therapy. We are reporting our experience of successful management of one such case of carcinoid crisis which was precipitated by Lu therapy.

Molecular insights into the mechanisms of susceptibility of Labeo rohita against oomycete Aphanomyces invadans.

Aphanomyces invadans, the causative agent of epizootic ulcerative syndrome, is one of the most destructive pathogens of freshwater fishes. To date, the disease has been reported from over 160 fish species in 20 countries and notably, this is the first non-salmonid disease that has resulted in major impacts globally. In particular, Indian major carps (IMCs) are highly susceptible to this disease. To increase our knowledge particularly with regards to host immune response against A. invadans infection in a susceptible host, the gene expression profile in head kidney of A. invadans-infected and control rohu, Labeo rohita was investigated using RNA sequencing. Time course analysis of RNA-Seq data revealed 5608 differentially expressed genes, involved among others in Antigen processing and presentation, Leukocyte transendothelial migration, IL-17 signaling, Chemokine signaling, C-type lectin receptor signaling and Toll-like receptor signaling pathways. In the affected pathways, a number of immune genes were found to be downregulated, suggesting an immune evasion strategy of A. invadans in establishing the infection. The information generated in this study offers first systematic mechanistic understanding of the host-pathogen interaction that might underpin the development of new management strategies for this economically devastating fish-pathogenic oomycete A. invadans.

Comparison of gustatory function between mucosal and squamous disease: a randomised controlled study.

OBJECTIVE: The aim of this study was to evaluate and compare the gustatory function between patients with chronic mucosal and squamous diseases before and after the surgery. METHOD: A total of 33 patients with mucosal diseases and 34 patients with squamous diseases were evaluated for gustatory function both in the pre-operative and post-operative periods. The taste scores were compared between the two groups. RESULTS: The gustatory scores in the mucosal disease group were significantly better than the scores in the squamous disease group (p < 0.05). There was no significant correlation detected between the age of the patients or duration of the disease with the taste scores in any of the study groups. CONCLUSION: The improvement in gustatory score was better in the chronic mucosal disease group than the squamous disease group. There was no significant correlation found between the age of the patients or duration of disease and the taste score in any of the study groups.

Hypomimia in Parkinson's disease: an axial sign responsive to levodopa.

BACKGROUND AND PURPOSE: Hypomimia is a prominent clinical feature in people with Parkinson's disease (PD), but it remains under-investigated. We aimed to examine the clinical correlates of hypomimia in PD and to determine whether this is a levodopa-responsive sign. METHODS: We included 89 people with PD. Hypomimia was assessed from digital video recordings by movement disorder specialists. Clinical evaluation included use of the Unified Parkinson's Disease Rating Scale part III (UPDRS-III), and assessment of motor and non-motor symptoms using standardized clinical scales. The relationships between hypomimia and other clinical data were analysed using Mann-Whitney U-tests and regression analysis. RESULTS: Hypomimia occurred in up to 70% of patients with PD. Patients with hypomimia had worse UPDRS-III 'off-medication' scores, mainly driven by bradykinesia and rigidity subscores. Patients with hypomimia also had worse apathy than patients without hypomimia. Finally, we found that hypomimia was levodopa-responsive and its improvement mirrored the change by levodopa in axial motor symptoms. CONCLUSION: Our study provides novel information regarding the clinical correlates of hypomimia in people with PD. A better understanding of hypomimia may be relevant for improving treatment and quality of life in PD.

Basal cell carcinoma arising over lesion of discoid lupus erythematosus: a rare occurrence.

Although the association of discoid lupus erythematosus (DLE) with squamous cell carcinoma has been described in the literature, coexistence with a basal cell carcinoma is very rare. The indolent and non-pruritic nature of the lesion cause it often to be diagnosed at the late stage of the disease. Long-standing photosensitivity and chronic inflammation later lead to scarring and hypopigmentation, which are considered as the risk factors for the malignant changes over DLE. Incisional biopsy is often performed from the erythematosus lesion which does not respond to medical treatment. Here we have reported a rare case of basal cell carcinoma of the skin developing in the background of long-standing DLE, which was successfully managed with cold knife local excision.

Ontogeny and tissue-specific expression of immune-relevant genes in Catla catla (Hamilton).

India is the second largest fish producing country in the world with production of 12.6 million tonnes (mt) in 2017-18, and Indian major carps (IMCs) contribute bulk of this fish production. Catla, Catla catla is the fastest growing species among IMCs. However, the survival rate of catla during larval rearing is normally lower than the other IMCs i.e rohu Labeo rohita, and mrigal Cirrhinus mrigala. Continuous efforts are devoted for the identification of nutritional and environmental requirements of fish larvae in order to reduce hatchery mortalities. However, very little information is available regarding physiology of the immune system, especially during the late larval and juvenile stages. Hence, understanding the ontogenetic development of immune-relevant genes in the larval stages of catla will serve as the markers for the development of immune competence and thereby, will be beneficial in developing effective immune intervention strategies. In the present study, expression profiles of some of the important innate (IL-1ß, IL-10, iNOS and C3) and adaptive immune (RAG-1, Ikaros, IgM and IgZ) genes during ontogenetic developmental stages and in different tissues of catla were investigated using quantitative real-time PCR. The results revealed that immune genes IL-1ß, C3, IgM and IgZ were detected in the unfertilized eggs indicating their maternal inheritance. Immune genes, IL-1ß, IL-10 and iNOS were expressed significantly during initial larval developmental stages whereas C3, RAG-1, Ikaros, IgM and IgZ showed significant expression during advanced stages of larval development in catla i.e. from 23 days post hatch (dph). Study of tissue distribution pattern of the genes indicated that innate immune genes were ubiquitously expressed in different tissues with varying degree of expression, whereas adaptive immune genes were predominantly expressed in lymphoid organs of catla. The information thus generated will improve knowledge on the maturation of the immune system in catla and will aid in deciding the appropriate age for vaccination in this teleost species.

Expression of immune genes in Indian major carp, Catla catla challenged with Flavobacterium columnare.

Columnaris disease, caused by Flavobacterium columnare, is one of the important bacterial diseases responsible for large-scale mortalities in numerous freshwater fishes globally. This disease can cause up to 100% mortality within 24 h of infection and is considered to be a cause of concern for aquaculture industry. Despite being a serious disease, scarce information is available regarding host-pathogen interaction, particularly the modulation of different immune genes in response to F. columnare infection. Therefore, in the present study, an attempt has been made to study expression of important immune regulatory genes, namely IL-1ß, iNOS, INF-γ, IL-10, TGF-ß, C3, MHC-I and MHC-II in gills and kidney of Catla catla following experimental infection with F. columnare. The expression analysis of immune genes revealed that transcript levels of IL-1ß, iNOS, IL-10, TGF-ß, C3 and MHC-I were significantly up-regulated (p < 0.05) in both the organs of the infected catla. IFN-γ and MHC-II were up-regulated in gills of infected catla whereas, both the genes showed down-regulation in kidney. The results indicate that important immune genes of C. catla are modulated following infection with F. columnare. The knowledge thus generated will strengthen the understanding of molecular pathogenesis of F. columnare in Indian major carp C. catla.

The feasibility and safety of graded adenosine stress test for myocardial perfusion in asthmatic and/or COPD patients.

SUBJECT AND METHODS: A total of 40 patients (M:F::26:14; age range: 37-84yrs; mean: 64.1yrs) with known chronic obstructive pulmonary disease (COPD) (ranging from mild to severe), referred for a stress myocardial perfusion study, were included in this study over a period of one year. All patients underwent adenosine stress in a titrated protocol and pre-infusion of short acting bronchodilator salbutamol 2 puffs few minutes prior to start adenosine infusion. In a fraction of 26 patients, pulmonary function tests (PFT) were performed and used in addition to clinical examination to classify the severity of pulmonary obstruction. On the basis of forced expiratory volume in one second (FEV1) on PFT, 4 patients had a mild disease (FEV1 60%-80%), 17 had a moderate obstructive disease (FEV1 41%-59%) and 4 had severe COPD/asthma (FEV1 <40%) while 2 patients had normal >95% FEV1. Post-stress questionnaire to assess subjective tolerance and symptoms were undertaken for all patients. RESULTS: The results demonstrated an excellent tolerance to adenosine infusion in this group of patients, with adequate stress achieved in all. None had complaints of severe dyspnoea or respiratory distress requiring medical intervention. Thirteen patients had mild to moderate degree dyspnoea during infusion. The study included a significant number of 23 elderly patients (>65 years), who showed better tolerance than the younger patients. CONCLUSION: In this pilot study in patients with COPD who referred for myocardial perfusion scintigraphy, the feasibility and safety of adenosine in a graded protocol along with a good pre-stress assessment and a short acting bronchodilator treatment was documented.