Inhibition of growth and virulence of Vibrio cholerae by carvacrol, an essential oil component of Origanum spp.

AIMS: In the age where bacterial resistance to conventional antibiotics is increasing at an alarming rate, the use of the traditional plant, herb extracts or other bioactive constituents is gradually becoming popular as an anti-virulence agent to treat pathogenic diseases. Carvacrol, a major essential oil fraction of Oregano, possesses a wide range of bioactivities. Therefore, we aimed to study the effect of sub-inhibitory concentrations of carvacrol on major virulence traits of Vibrio cholerae. METHODS AND RESULTS: We have used in vitro as well as ex vivo models to access the anti-pathogenic role of carvacrol. We found that the sub-inhibitory concentration of carvacrol significantly repressed bacterial mucin penetrating ability. Carvacrol also reduced the adherence and fluid accumulation in the rabbit ileal loop model. Reduction in virulence is associated with the downregulated expression of tcpA, ctxB, hlyA and toxT. Furthermore, carvacrol inhibits flagellar synthesis by downregulating the expression of flrC and most of the class III genes. CONCLUSIONS: Carvacrol exhibited anti-virulence activity against V. cholerae, which involved many events including the inhibition of mucin penetration, adhesion, reduced expression of virulence-associated genes culminating in reduced fluid accumulation. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that carvacrol possesses inhibitory activity against V. cholerae pathogenesis and might be considered as a potential bio-active therapeutic alternative to combat cholera.

Exploring the physical and quality attributes of muffins incorporated with microencapsulated squalene as a functional food additive.

Squalene, a triterpenoid compound is proven to possess immense bioactivities by virtue of its high antioxidant activity. The present study was designed to investigate the quality attributes of muffins as influenced by addition of encapsulated squalene. Nutritional analysis showed that calorific value of prepared muffins has ranged from 480.78 ± 0.10 to 501.61 ± 0.38 kcal. Baking loss was lowest in case of muffins prepared with encapsulated squalene with its crumb region recorded higher moisture content. Color kinetics study indicated that browning index (BI) was higher in crust portion of encapsulated squalene enriched muffins. Scanning electron micrographs showing that muffins with encapsulated squalene had stronger structural organization. This was further supported by the textural studies showed that the muffins with encapsulated squalene was cohesive, springier and chewy with less gumminess and stiffness indicating their efficacy in improving the textural quality. Oxidative stability and microbiological quality were also high in squalene enriched foods suggesting that squalene might have some antimicrobial effects. Outcome of the study indicated that encapsulated squalene can be very well utilised as a functional food ingredient in ready -to-eat functional foods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-020-04955-9).

Identification of common virulence factors present in enterotoxigenic Escherichia coli isolated from diarrhoeal patients in Kolkata, India.

AIMS: Development of an effective vaccine against enterotoxigenic Escherichia coli (ETEC) is largely dependent on the conscientious understanding of different virulence associated factors from diverse geographical areas. So, the objective of this study is to elucidate the distribution of enterotoxins, CF and NCVF in clinical ETEC strains isolated between 2008 and 2014 from two hospitals in Kolkata, India. METHODS AND RESULTS: Multiplex PCR method was used for detection of two enterotoxin genes, 11 common CFs and five common NCVFs. Among the 350 tested ETEC strains, 61% strains possessed est+elt genes, 25% est and 14% elt. Among 56% CF positive ETEC strains, CS21 was the prevalent one (37%) followed by CS6 (36%). NCVF genes were present in 59% of the ETEC strains; eatA was the most prevalent (65%) followed by etpA (51%). There were 29% strains negative for any CFs or NCVFs. CONCLUSIONS: We conclude that a pattern exists between CS6, eatA and toxins. We observed est with or without elt, CS6 with or without CS5 and with or without eatA were present in 24% of clinical ETEC strains (59/250) analysed. CS21 has emerged as another predominant CF but it had diverse CFs and NCVFs. SIGNIFICANCE AND IMPACT OF THE STUDY: Prevalence of ETEC virulence factors would help in tracking ETEC globally and suggests the need of a multivalent ETEC vaccine.

Prediction of clinical outcome in patients treated with cardiac resynchronization therapy - the role of NT-ProBNP and a combined response score.

BACKGROUND: Cardiac resynchronization therapy (CRT) is an established therapy for appropriately selected patients with heart failure. Response to CRT has been heterogeneously defined using both clinical and echocardiographic measures, with poor correlation between the two. METHODS: The study cohort was comprised of 202 CRT-treated patients and CRT response was defined at 6 months post-implant. Echocardiographic response (E+) was defined as a reduction in LVESV ≥ 15%, clinical response as an improvement of ≥ 1 NYHA class (C+), and biomarker response as a ≥ 25% reduction in NT-proBNP(B+). The association of response measures (E+, B+, C+; response score range 0-3) and clinical endpoints at 3 years was assessed in landmarked Cox models. RESULTS: Echo and clinical responders demonstrated greater declines in NT-proBNP than non-responders (median [E+/B+]: -52%, [E+]: -27%, [C+]: -39% and [E-/C-]: -13%; p = 0.01 for trend). Biomarker (HR 0.43 [95% CI: 0.22-0.86], p = 0.02) and clinical (HR 0.40 [0.23-0.70] p = 0.001) response were associated with a significantly reduced risk of the primary endpoint. When integrating each response measure into a composite score, each 1 point increase was associated with a 31% decreased risk for a composite endpoint of mortality, LVAD, transplant and HF hospitalization (HR 0.69 [95% CI: 0.50-0.96], p = 0.03), and a 52% decreased risk of all-cause mortality (HR 0.48 [95% CI: 0.26-0.89], p = 0.02). CONCLUSION: Serial changes in NT-proBNP are associated with clinical outcomes following CRT implant. Integration of biomarker, clinical, and echocardiographic response may discriminate CRT responders versus non-responders in a clinically meaningful way, and with higher accuracy. TRIAL REGISTRATION: The cohort was combined from study NCT01949246 and the study based on local review board approval 2011/550 in Lund, Sweden.

Antioxidant, functional properties and amino acid composition of pepsin-derived protein hydrolysates from whole tilapia waste as influenced by pre-processing ice storage.

In recent years, hygienic handling of fishery waste is demanded owing to the fact that the fishery waste is an ideal raw material for the preparation of bioactive compounds. In the present study, the effect of pre-processing storage (at 4 ± 2 °C) of whole tilapia waste (WTW) on the properties of its protein hydrolysate derived using pepsin was evaluated. Fish protein hydrolysates (FPH) were prepared from 0, 24 and 48 h old ice stored WTW and designated as FPH-0, FPH-1, and FPH-2, respectively. Total amino acids, total essential amino acids and total hydrophobic amino acids of FPH samples increased with the storage period of raw material (WTW). Antioxidant activities such as DPPH (2, 2 diphynyl-1-picrylhydrazyl) free radical scavenging activity and ferric reducing power of FPH samples were dose dependent. FPH-0 had better antioxidant properties including linoleic acid peroxidation inhibition activity than FPH-1 and FPH-2. The DNA nicking assay revealed the protective effect of FPH preparations against Fenton's reaction mediated oxidative damage. FPH-2 had better emulsifying properties and foaming stability whereas the FPH-0 had relatively good foaming capacity. SDS-PAGE indicated the presence of peptides ranging from 116 to < 14.4 kDa in FPH-0 and less than 18 kDa in FPH-1 and FPH-2. The present study, clearly demonstrated that whole tilapia waste can effectively be converted to FPH and could be a potential ingredient in functional food and as a rich source of high-quality protein in animal feed formulations.

Kinematic variables and water transport control the formation and location of arc volcanoes.

The processes that give rise to arc magmas at convergent plate margins have long been a subject of scientific research and debate. A consensus has developed that the mantle wedge overlying the subducting slab and fluids and/or melts from the subducting slab itself are involved in the melting process. However, the role of kinematic variables such as slab dip and convergence rate in the formation of arc magmas is still unclear. The depth to the top of the subducting slab beneath volcanic arcs, usually approximately 110 +/- 20 km, was previously thought to be constant among arcs. Recent studies revealed that the depth of intermediate-depth earthquakes underneath volcanic arcs, presumably marking the slab-wedge interface, varies systematically between approximately 60 and 173 km and correlates with slab dip and convergence rate. Water-rich magmas (over 4-6 wt% H(2)O) are found in subduction zones with very different subduction parameters, including those with a shallow-dipping slab (north Japan), or steeply dipping slab (Marianas). Here we propose a simple model to address how kinematic parameters of plate subduction relate to the location of mantle melting at subduction zones. We demonstrate that the location of arc volcanoes is controlled by a combination of conditions: melting in the wedge is induced at the overlap of regions in the wedge that are hotter than the melting curve (solidus) of vapour-saturated peridotite and regions where hydrous minerals both in the wedge and in the subducting slab break down. These two limits for melt generation, when combined with the kinematic parameters of slab dip and convergence rate, provide independent constraints on the thermal structure of the wedge and accurately predict the location of mantle wedge melting and the position of arc volcanoes.

Genome-wide association study meta-analysis of European and Asian-ancestry samples identifies three novel loci associated with bipolar disorder.

Meta-analyses of bipolar disorder (BD) genome-wide association studies (GWAS) have identified several genome-wide significant signals in European-ancestry samples, but so far account for little of the inherited risk. We performed a meta-analysis of ∼750,000 high-quality genetic markers on a combined sample of ∼14,000 subjects of European and Asian-ancestry (phase I). The most significant findings were further tested in an extended sample of ∼17,700 cases and controls (phase II). The results suggest novel association findings near the genes TRANK1 (LBA1), LMAN2L and PTGFR. In phase I, the most significant single nucleotide polymorphism (SNP), rs9834970 near TRANK1, was significant at the P=2.4 × 10(-11) level, with no heterogeneity. Supportive evidence for prior association findings near ANK3 and a locus on chromosome 3p21.1 was also observed. The phase II results were similar, although the heterogeneity test became significant for several SNPs. On the basis of these results and other established risk loci, we used the method developed by Park et al. to estimate the number, and the effect size distribution, of BD risk loci that could still be found by GWAS methods. We estimate that >63,000 case-control samples would be needed to identify the ∼105 BD risk loci discoverable by GWAS, and that these will together explain <6% of the inherited risk. These results support previous GWAS findings and identify three new candidate genes for BD. Further studies are needed to replicate these findings and may potentially lead to identification of functional variants. Sample size will remain a limiting factor in the discovery of common alleles associated with BD.

Common genetic variants in the 9p21 region and their associations with multiple tumours.

BACKGROUND: The chromosome 9p21.3 region has been implicated in the pathogenesis of multiple cancers. METHODS: We systematically examined up to 203 tagging SNPs of 22 genes on 9p21.3 (19.9-32.8 Mb) in eight case-control studies: thyroid cancer, endometrial cancer (EC), renal cell carcinoma, colorectal cancer (CRC), colorectal adenoma (CA), oesophageal squamous cell carcinoma (ESCC), gastric cardia adenocarcinoma and osteosarcoma (OS). We used logistic regression to perform single SNP analyses for each study separately, adjusting for study-specific covariates. We combined SNP results across studies by fixed-effect meta-analyses and a newly developed subset-based statistical approach (ASSET). Gene-based P-values were obtained by the minP method using the Adaptive Rank Truncated Product program. We adjusted for multiple comparisons by Bonferroni correction. RESULTS: Rs3731239 in cyclin-dependent kinase inhibitors 2A (CDKN2A) was significantly associated with ESCC (P=7 × 10(-6)). The CDKN2A-ESCC association was further supported by gene-based analyses (Pgene=0.0001). In the meta-analyses by ASSET, four SNPs (rs3731239 in CDKN2A, rs615552 and rs573687 in CDKN2B and rs564398 in CDKN2BAS) showed significant associations with ESCC and EC (P<2.46 × 10(-4)). One SNP in MTAP (methylthioadenosine phosphorylase) (rs7023329) that was previously associated with melanoma and nevi in multiple genome-wide association studies was associated with CRC, CA and OS by ASSET (P=0.007). CONCLUSION: Our data indicate that genetic variants in CDKN2A, and possibly nearby genes, may be associated with ESCC and several other tumours, further highlighting the importance of 9p21.3 genetic variants in carcinogenesis.

Aspirin but not ibuprofen use is associated with reduced risk of prostate cancer: a PLCO study.

BACKGROUND: Although most epidemiological studies suggest that non-steroidal anti-inflammatory drug use is inversely associated with prostate cancer risk, the magnitude and specificity of this association remain unclear. METHODS: We examined self-reported aspirin and ibuprofen use in relation to prostate cancer risk among 29 450 men ages 55-74 who were initially screened for prostate cancer from 1993 to 2001 in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Men were followed from their first screening exam until 31 December 2009, during which 3575 cases of prostate cancer were identified. RESULTS: After adjusting for potential confounders, the hazard ratios (HRs) of prostate cancer associated with <1 and ≥ 1 pill of aspirin daily were 0.98 (95% confidence interval (CI), 0.90-1.07) and 0.92 (95% CI: 0.85-0.99), respectively, compared with never use (P for trend 0.04). The effect of taking at least one aspirin daily was more pronounced when restricting the analyses to men older than age 65 or men who had a history of cardiovascular-related diseases or arthritis (HR (95% CI); 0.87 (0.78-0.97), 0.89 (0.80-0.99), and 0.88 (0.78-1.00), respectively). The data did not support an association between ibuprofen use and prostate cancer risk. CONCLUSION: Daily aspirin use, but not ibuprofen use, was associated with lower risk of prostate cancer risk.

Phlorotannins-bioactivity and extraction perspectives.

Phlorotannins, a seaweed based class of polyphenolic compounds, have proven to possess potential bioactivities such as antioxidant, antimicrobial, anti-allergic, anti-diabetic, anti-inflammatory, anti-cancerous, neuroprotection etc. These bioactivities have further increased demand globally and sustainable techniques such as supercritical fluid extraction, microwave assisted extraction, enzyme assisted extraction, extraction using deep eutectic solvents etc. are being explored currently for production of phlorotannin-rich extracts. In spite of such well documented bioactivities, very few phlorotannin-based nutraceuticals are available commercially which highlights the significance of generating consumer awareness about their physiological benefits. However, for industry level commercialization accurate quantification of phlorotannins with respect to the different classes is vital requiring sophisticated analytical techniques such as mass spectrometry, 1H-NMR spectroscopy etc. owing to the wide structural diversity. This review summarizes the extraction and bioactivities of phlorotannins based on the findings of in vivo and in vitro studies.

Efficacy of intra-articular dexmedetomidine for postoperative analgesia in arthroscopic knee surgery.

INTRODUCTION: Various drugs are administered intraarticularly to provide postoperative analgesia after arthroscopic knee surgery, of which opioids and alpha2 agonists could be of particular interest. METHODS: Sixty patients undergoing elective knee arthroscopy were randomly assigned to two groups (n = 30). Group R received 19 ml of 0.25% ropivacaine and 1 ml of isotonic saline (total volume 20 ml) intraarticularly. Group RD received 100 microg (1 ml) of dexmedetomidine added to 19 ml of 0.25% ropivacaine intra-articularly (total volume 20 ml). Analgesic effect was evaluated by measuring pain intensity (VAS score) and duration of analgesia. RESULTS: A longer delay was observed between intraarticular injection of study medication and first requirement of supplementary analgesic in group RD (10.84 +/- 2.6 hours) compared to group R (5.38 +/- 1.4 hours). Total consumption of fentanyl citrate in postoperative period was significantly less in group RD. No significant side effects were noted. CONCLUSION: Dexmedetomidine, added as adjunct to ropivacaine in patients undergoing arthroscopic knee surgery, improve the quality and duration of postoperative analgesia.

Human epidermal growth factor receptor-2 and estrogen receptor expression, a demonstration project using the residual tissue repository of the Surveillance, Epidemiology, and End Results (SEER) program.

BACKGROUND: In 2001, the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) program established Residual Tissue Repositories (RTR) in the Hawaii, Iowa, and Los Angeles Tumor Registries to collect discarded tissue blocks from pathologic laboratories within their catchment areas. To validate the utility of the RTR for supplementing SEER's central database, we assessed human epidermal growth factor receptor-2 (HER2) and estrogen receptor expression (ER) in a demonstration project. MATERIALS: Using a prepared set of tissue microarrays (TMAs) residing in the Hawaii Tumor Registry (HTR), we performed standard immunohistochemistry. Breast cancers in the TMA were diagnosed in 1995, followed through 2006, and linked to SEER's main database. RESULTS: The TMA included 354 cases, representing 51% of 687 breast cancers in the HTR (1995). The HTR and TMA cases were similar with respect to patient demographics and tumor characteristics. Seventy-six percent (76%, 268 of 354) of TMA cases were HER2+ and/or ER+, i.e., 28 HER2+ER-, 12 HER2+ER+, and 228 HER2-ER+. There were 67 HER2-ER- cases and 19 were unclassified. Age distributions at diagnosis were bimodal with dominant early-onset modes for HER2+ER- tumors and dominant late-onset modes for HER2-ER+ breast cancers. Epidemiologic patterns for concordant HER2+ER+ (double-positive) and HER2-ER- (double-negative) were intermediate to discordant HER2+ER- and HER2-ER+. CONCLUSION: Results showed contrasting incidence patterns for HER2+ (HER2+ER-) and ER+ (HER2-ER+) breast cancers, diagnosed in 1995. Though sample sizes were small, this demonstration project validates the potential utility of the RTR for supplementing the SEER program.

A low-GC Gram-positive Thermoanaerobacter-like bacterium isolated from an Indian hot spring contains Cr(VI) reduction activity both in the membrane and cytoplasm.

AIM: Characterization of an anaerobic thermophilic bacterium and subcellular localization of its Cr(VI)-reducing activity for potential bioremediation applications. METHODS AND RESULTS: 16S rRNA gene sequence-based analyses of bacterial strains isolated from sediment samples of a Bakreshwar (India) hot spring, enriched anaerobically in iron-reducing medium, found them to be 86-96% similar to reported Thermoanaerobacter strains. The most efficient iron reducer among these, BSB-33, could also reduce Cr(VI) at an optimum temperature of 60 degrees C and pH 6.5. Filtered culture medium could reduce Cr(VI) but not Fe(III). Cell-free extracts reduced Cr(VI) inefficiently under aerobic conditions but efficiently anaerobically. Fractionation of the cell-free extracts showed that chromium reduction activity was present in both the cytoplasm and membrane. CONCLUSIONS: BSB-33 reduced Fe(III) and Cr(VI) anaerobically at 60 degrees C optimally. After fractionation, the reducing activity of Cr(VI) was found in both cytoplasmic and membrane fractions. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first systematic study of anaerobic Cr(VI) reduction by a gram-positive thermophilic micro-organism and, in contrast to our results, none of the earlier reports has mentioned Cr(VI)-reducing activity to be present both in the cytoplasm and membrane of an organism. The strain may offer itself as a potential candidate for bioremediation.

Prosthetic replacement of elbow in postburn bony ankylosis: long-term results.

Fifteen patients with 17 post-burn bony ankylosed elbows (two bilateral) fixed at a mean of 72.5 degrees +/- 27.5 degrees flexion were treated with Baksi sloppy hinge elbow arthroplasty. All patients agreed to avoid strenuous use of their replaced elbows. There were four males and 11 females whose ages varied from 21 to 48 years (average 31.6 years). One of the five patients who had severe anterior skin contracture required release. One patient died during the early follow-up period. During the follow-up studies which ranged from 11 to 23 years (average 14.1), eleven patients (68.7%) regained painless stable motion with mean recovery of arc of flexion from 20 degrees +/-10 degrees to 107.5 degrees +/-17.5 degrees with mean range of flexion 90 degrees +/-20 degrees . Good forearm motion was retained in all cases. According to the assessment score (Morrey BF, The elbow and it's disorder, 2nd edn. W.B. Saunders, Philadelphia, 1993), nine (56.2%) patients were recorded as excellent, two (12.5%) as good, and five (31.5%) as poor who were considered as failures. The five failures required removal of the prosthesis but retained acceptable elbow function due to periprosthetic fibrosis connecting the adjacent bone ends.

Assaying baseline status of particulate laden polyaromatic hydrocarbon for a grass root level industrial project.

A study of particulate laden polyaromatic hydrocarbon was conducted at 13 selected locations in a 10 km radial distance of a proposed site for a grass root level industry. Suspended particulate matter samples were continuously monitored for 24 h over a period of 3 months. The Polyaromatic Hydrocarbons (PAHs) were extracted from the particulate samples and analysed using Gas Chromatograph-Mass Spectrometer. Limit of Quantification was also established for individual PAHs. Coal combustion and traffic emission were the major contributors for PAHs in the region. The relative contribution of 2, 3, 4, 5, and 6 ring PAHs in particulates of different sampling sites was also investigated and it is observed that 4 ring (29.76%) and 5 ring (29.06%) compounds are prominent in the particulates measured in the region.

Chitosan - Whey protein as efficient delivery system for squalene: Characterization and functional food application.

Squalene, a triterpenoid compound possessing excellent bioactivities, is not being utilized as a functional food ingredient due to its high susceptibility to oxidation. In the present study, the feasibility of chitosan-whey protein as an efficient wall material for squalene encapsulation using spray drying technique was attempted for functional food applications. The encapsulation efficiency of the squalene powder was found to be 75.4 ±â€¯0.22% whereas other physico-chemical properties such as moisture content, flowability, solubility, peroxide value, etc. have shown satisfactory results. The thermogravimetric analysis showed that chitosan-whey protein was able to retain the thermal stability of squalene up to a temperature of 422 °C. Furthermore, the functional food application of the encapsulated squalene in a bakery product (cake) exhibited significantly (p < 0.05) better properties in terms of oxidative stability, sensory attributes than that of cake with pure squalene and control treatment. Hence, it can be concluded that emulsification of squalene in chitosan-whey protein and its subsequent encapsulation by spray drying can be a potential process to produce oxidatively stable encapsulates for the development of functional foods.

Vitamin D receptor polymorphisms and renal cancer risk in Central and Eastern Europe.

Previous studies investigated the role of vitamin D intake and cancer risk. The kidney is a major organ for vitamin D metabolism, activity, and calcium homeostasis; therefore, it was hypothesized that dietary vitamin D intake and polymorphisms in the vitamin D receptor (VDR) gene may modify renal cell carcinoma (RCC) risk. Three common VDR gene polymorphisms (BsmI, FokI, TaqI) were evaluated among 925 RCC cases and 1192 controls enrolled in a hospital-based case-control study conducted in Central and Eastern Europe. Overall associations with RCC risk were not observed; however, subgroup analyses revealed associations after stratification by median age of diagnosis and family history of cancer. Among subjects over 60 yr, reduced risks were observed among carriers of the f alleles in the FokI single-nucleotide polymorphism (SNP) (odds ratio [OR] = 0.61 for Ff and OR = 0.74 for ff genotypes) compared to subjects with the FF genotype (P trend = 0.04; P interaction = 0.004). Subjects with the BB BsmI genotype and a positive family history of cancer had lower risk compared to subjects with the bb allele (OR = 0.60; 95% CI: 0.33-1.1; P trend = 0.05). Genotype associations with these subgroups were not modified when dietary sources of vitamin D or calcium were considered. Additional studies of genetic variation in the VDR gene are warranted.

How predictive is the Framingham's risk prediction algorithm in Indian perspective? A retrospective case-control study from Kolkata.

OBJECTIVE: In this retrospective case-control study, an attempt was made to assess the predictive efficacy of Framingham's risk prediction algorithm in Indian perspective. METHODS: A total of 350 patients and 293 age- and sex-matched controls were considered in the study. Those patients, who were presenting for the first time with acute coronary syndrome (ACS) and who did not have any prior manifestation of coronary heart disease (CHD) formed the patient group. The risk prediction algorithm was applied to obtain the risk score and the corresponding 10-year risk in each patient and control. They were divided into two groups: diabetic and nondiabetic. Depending on the 10-year risk, they were further grouped into high risk (10-year risk > 20%), moderately high risk (10-year risk between 10% to 20%), and low risk (10-year risk < 10%). The results were compared and statistically analyzed. RESULTS: In the diabetic patients with ACS, 14.29% qualified as high risk, 32.79% as moderately high risk, and 52.94% as low risk. The corresponding figures for diabetic subjects without ACS were 3.26%, 54.35%, and 42.39%, respectively. In nondiabetic patients with ACS, 19.91% were in the high-risk group, 38.96% in moderately high risk, and 41.13% in the low-risk group; while among the controls, the corresponding figures were 9.95%, 21.89%, and 68.16%, respectively. In nondiabetic subjects, the mean risk was significantly higher for patients compared to controls (14.13 vs. 8.61, p < 0.01). However, in diabetic subjects, there was no significant difference in the mean projected risk between those with ACS and those without ACS (11.37 vs. 10.41, p = NS). CONCLUSION: In the Indian perspective, Framingham's risk prediction protocol has a fair amount of predictive efficacy since the difference of mean risk score between the patients and controls was statistically significant. However, it fails to identify a large proportion of high-risk nondiabetic patients. Hence, a better protocol for the Indian perspective is badly needed.