Development of Novel Indazolyl-Acyl Hydrazones as Antioxidant and Anticancer Agents that Target VEGFR-2 in Human Breast Cancer Cells.

The increased expression of VEGFR-2 in a variety of cancer cells promotes a cascade of cellular responses that improve cell survival, growth, and proliferation. Heterocycles are common structural elements in medicinal chemistry and commercially available medications that target several biological pathways and induce cell death in cancer cells. Herein, the evaluation of indazolyl-acyl hydrazones as antioxidant and anticancer agents is reported. Compounds 4e and 4j showed inhibitory activity in free radical scavenging assays (DPPH and FRPA). The titled compounds were employed in cell viability studies using MCF-7 cells, and it was observed that compounds 4f and 4j exhibited IC50 values 15.83 µM and 5.72 µM, respectively. In silico docking revealed the favorable binding energies of -7.30 kcal/mol and -8.04 kcal/mol for these compounds towards Vascular Endothelial Growth Factor Receptor-2 (VEGFR-2), respectively. In conclusion, compounds with antioxidant activity and that target VEGFR-2 in breast cancer cells are reported.

Preparation of new organo-ruthenium(II) complexes and their nucleic acid/albumin binding efficiency and in vitro cytotoxicity studies.

Hetero-bimetallic ruthenium(II) complexes (PRAFIZ and PRBFIZ) containing acetyl ferrocene (AFIZ)/benzoyl ferrocene isonicotinic hydrazone ligands (BFIZ) were synthesized and characterized by various spectral and analytical techniques. The structure of acetyl ferrocene isonicotinic hydrazone (AFIZ) and the complex PRBFIZ was confirmed by X-ray crystallography. The hydrazide ligands coordinated in a bidentate monobasic fashion using their N1 hydrazinic nitrogen and enolic oxygen atoms. The binding interactions of the ligands and complexes were examined using Calf-Thymus DNA (CT-DNA) and bovine serum albumin (BSA). Scanning Electron Microscopic (SEM) experiments clarified the efficient binding interaction of the ligands and complexes with BSA. The results of in vitro cytotoxicity studies on MDA-MB-261 breast cancer cells and A549 human lung cancer cells and cell morphological analysis results through staining assays clearly indicated the cytotoxic nature of the complexes.

COVID-19 disease in peritoneal dialysis patients: a single centre experience from India.

BACKGROUND: This study presents our data on mortality in end stage renal disease (ESRD) patients on peritoneal dialysis (PD) who developed COVID-19. MATERIALS AND METHODS: Sri Padmavathi Medical College Hospital, Sri Venkateswara Institute of Medical Sciences University, was designated the State COVID Hospital in March 2020. In a retrospective observational study, we collected the data of ESRD patients on PD and identified the risk factors for mortality. RESULTS: Prior to the pandemic, 136 patients with ESRD were on peritoneal dialysis at our Institute. Among them, 27 (19.8%) eventually developed COVID-19, and 14 of them (51.8%) died. Serum albumin levels were lower and D-dimer levels were significantly higher in deceased patients than in survivors. DISCUSSION: The mortality rate in ESRD patients on PD with COVID-19 at our institution was higher than in other published studies.

Assessment of toxicities and outcomes in patients with breast cancer treated with hypofractionated radiotherapy.

BACKGROUND: Hypofractionation is now becoming the standard of care in breast irradiation. The aim of this study was to assess the toxicities and outcomes in patients with breast cancer treated with hypofractionated radiotherapy (HFRT). METHODS: Patients with localized breast cancer who received adjuvant HFRT between 2013 and 2015 with a minimum follow-up of 6 months following radiation were included in this prospective study. Late toxicities were assessed using CTCAE v 4 and included chest/breast pain, limb pain, limb edema, skin pigmentation, skin fibrosis, and shoulder movement restriction. Outcomes assessed included locoregional control, disease-free survival, and overall survival. Statistical analysis was done using Microsoft Excel and SPSS v22. RESULTS: A total of 81 patients fulfilled the inclusion criteria, of which 19 patients had died during follow-up. Regional nodal irradiation was done in 63 (77.8%) patients using the same hypofractionated schedule of 40 Gy in 15 fractions. Late toxicities were assessed for 62 patients. The median follow-up following the course of hypofractionated radiation was 45 months (range 14 - 65 months). Late toxicities were assessed for 62 patients. Grade 1/2 chest/breast pain, limb pain, limb edema, skin pigmentation, skin fibrosis, and shoulder movement restriction were seen in 11%, 12%, 7%, 6%, 8%, and 11% of cases, respectively. Distant recurrences were seen in 8% of cases, and there were no locoregional recurrences. Five-year overall survival was 76.5%. CONCLUSION: HFRT to whole breast or chest wall and the regional nodal areas was well-tolerated with acceptable rates of late toxicities on follow-up.

Ocular adverse events in drug sensitive TB patients on daily fixed dose combination anti-TB drugs: A record review study from Kerala, India.

BACKGROUND: Government of India's Revised National TB Control Programme (RNTCP) has begun implementing daily fixed dose combination (FDC) anti-TB treatment regimen for drug sensitive TB patients in which ethambutol is given for six months. Prolonged ethambutol use is known to cause ocular adverse drug events (ADE). OBJECTIVES: To assess the magnitude of ocular ADEs in adult drug sensitive TB patients initiated on daily FDCs and to describe the demographic and clinical profile of patients with ocular ADEs. METHODS: We conducted a retrospective cohort study involving review of RNTCP records of all adult (age >14 years) drug sensitive TB patients initiated on daily FDCs between1st January 2018 and 31st July 2018 in Thiruvananthapuram district, Kerala State, India. RESULTS: 714 patients were initiated on daily FDCs during the study period. It was unknown whether all patients had undergone assessment for ocular ADEs. However, of these 714 patients, 8 patients (1.1%) were documented to have had ocular ADEs. Seven of these 8 patients had received ethambutol more than 15 mg/kg body weight and had developed ocular symptoms (decreased/blurring of vision) 3 months after TB treatment initiation. Ethambutol was stopped in all these 8 patients. In 5 patients it was recorded that ocular ADEs had resolved following stoppage of ethambutol and in the remaining it was unknown. CONCLUSION: The study confirms the occurrence of ocular ADEs among drug sensitive TB patients on daily FDCs and recommends strengthening of systems for assessing, documenting and managing ocular ADE.

Modeling of linear programming and extended TOPSIS in decision making problem under the framework of picture fuzzy sets.

Picture fuzzy sets (PFSs) are comparatively a new extension of fuzzy sets which describe the human opinions that has more answers like acceptance, rejection, neutral and desist, which cannot be correctly presented in fuzzy sets (FSs) and intuitionistic fuzzy sets (IFSs). The PFSs are categorized by three objects, the degree of belonging, the degree of neutral belonging and the degree of non- belonging such that the total of these three degrees must not be more than one. So far, there is no such work presented in the literature which deals with unknown weights of criteria based on PFSs. In the present work, we have developed a linear programming (LP) model to find the exact weights from the given constraints of weights for the criteria and construct a modified distance based on similarity measure between picture fuzzy sets. Then we have utilized this similarity measure to achieve the best option in the multiple criteria decision making (MCDM) problem. Lastly, two practical examples for the selection of alternatives are presented to compare the obtained results with the existing similarity measures.

Immunohistochemical Detection of Proliferative Marker Ki-67 in Benign and Malignant Salivary Gland Tumors.

Introduction: Salivary gland tumors are the most histologically heterogeneous group of tumors with the greatest diversity of morphologic features among their cells and tissues. The present study was aimed at assessing the validity of Ki-67, a cell proliferation marker, as a prognostic factor in benign and malignant salivary gland tumors and to study whether it is related to age, sex, anatomical site, and size of the lesion in salivary gland tumors. Materials and methods: A retrospective study consisted of benign salivary gland tumors (BSGTs) (n = 15), malignant salivary gland tumors (n = 18), and normal salivary gland parenchyma (n = 15). Results: There was a significant difference of Ki-67 labeling index (LI, %) in normal salivary gland parenchyma, BSGTs, and malignant salivary gland tumors. The Ki-67 LI (%) in normal salivary gland parenchyma is negligible (0.27 ± 0.31%), whereas malignant salivary gland tumors showed very high Ki-67 LI (%) of 18.79 ± 18.06% compared with BSGTs being 0.76 ± 2.02%. There was a significant correlation statistically of mean ± standard deviation (SD) of Ki-67 LI (%) with the age of the patients being the maximum (32.68 ± 15.87%) in the 50 to 59 years age group, whereas sex, site of the lesion, and size of the lesion in salivary gland tumors had no significant correlation. Conclusion: The Ki-67 is a useful marker for assessing prolif-erative potential of tumors. Clinical significance: The Ki-67 LI% can be used as a reliable adjuvant diagnostic tool to differentiate between the subtypes and grading of certain malignant tumors, such as mucoepi-dermoid carcinoma (MEC), adenoid cystic carcinoma (AdCC), and acinic cell carcinoma (AcCC), which are usually difficult to diagnose on histopathological criteria alone. Keywords: Immunohistochemistry, Ki-67, Salivary gland neoplasms.

Matrix metalloproteinases-3 gene-promoter polymorphism as a risk factor in oral submucous fibrosis in an Indian population: a pilot study.

AIM: Host-related factors could be important to the fundamental understanding of oral diseases. One such factor is matrix metalloproteinases (MMP). MMP3 (stromelysin 1) is crucial in connective tissue remodeling. Promoter regions are specific segments of DNA that control the rate of mRNA synthesis. Thus, polymorphism in the (5A/6A) promoter region of the MMP3 gene results in different transcriptional activities and is related to susceptibility in diseases. The aim of the present study was to detect polymorphism in the MMP3 gene-promoter region in oral submucous fibrosis patients (OSF), oral squamous cell carcinoma patients (OSCC), and normal individuals with tobacco and areca nut habit and without lesions, and to correlate the genotype as a risk factor in these diseases. METHOD: Genomic DNA from blood of OSF (n = 5), OSCC (n = 5), and normal individuals with tobacco and areca nut habit (n = 5) and without (n = 5) were subjected to polymerase chain reaction of the MMP3 gene-promoter region and DNA sequencing. RESULTS: The 5A allele in the MMP3 gene-promoter region was observed more frequently in the OSF group than the control groups. No significant difference was noted between OSCC and the control groups on the 5A allele. CONCLUSION: The results indicated that the 5A allele of the MMP3 gene-promoter region could be associated with OSF risk factor, but not OSCC, in an Indian population.

Detection of Porphyromonas gingivalis and Treponema denticola in chronic and aggressive periodontitis patients: A comparative polymerase chain reaction study.

BACKGROUND: The detection frequency of Porphyromonas gingivalis and Treponema denticola in chronic periodontitis (CP) and aggressive periodontitis (AgP) is not explored well in Indian population. AIM: The study was undertaken to detect P. gingivalis and T. denticola in CP as well as in AgP patients using polymerase chain reaction (PCR), and to determine the relationship between the frequency of these two microorganisms and the severity of clinical periodontal parameters. MATERIALS AND METHODS: Subgingival plaque samples were collected from ninety participants (thirty CP patients, thirty AgP patients, and thirty healthy participants) and the aforementioned two microorganisms were detected using PCR. RESULTS: However, when CP and AgP were compared for the detection frequency of two microorganisms, no statistically significant difference was noted. A statistically significant increase in the number of bacteria-positive sites increased as the score of plaque index (PI), gingival index (GI), and clinical attachment level of CP and AgP patients increased. Coexistence of P. gingivalis and T. denticola was frequently observed in deep periodontal pockets. CONCLUSIONS: Study findings suggest that P. gingivalis and T. denticola are significantly associated with the severity of periodontal tissue destruction. Statistically significant association exists between clinical periodontal parameters such as PI, GI, periodontal pocket depth (PPD), and clinical attachment loss and presence of both the microorganisms.

Entomopathogenic Nematodes Combined with Adjuvants Presents a New Potential Biological Control Method for Managing the Wheat Stem Sawfly, Cephus cinctus (Hymenoptera: Cephidae).

The wheat stem sawfly, (Cephus cinctus Norton) Hymenoptera: Cephidae, has been a major pest of winter wheat and barley in the northern Great Plains for more than 100 years. The insect's cryptic nature and lack of safe chemical control options make the wheat stem sawfly (WSS) difficult to manage; thus, biological control offers the best hope for sustainable management of WSS. Entomopathogenic nematodes (EPNs) have been used successfully against other above-ground insect pests, and adding adjuvants to sprays containing EPNs has been shown to improve their effectiveness. We tested the hypothesis that adding chemical adjuvants to sprays containing EPNs will increase the ability of EPNs to enter wheat stems and kill diapausing WSS larvae. This is the first study to test the ability of EPNs to infect the WSS, C. cinctus, and test EPNs combined with adjuvants against C. cinctus in both the laboratory and the field. Infection assays showed that three different species of EPNs caused 60-100% mortality to WSS larvae. Adding Penterra, Silwet L-77, Sunspray 11N, or Syl-Tac to solutions containing EPNs resulted in higher WSS mortality than solutions made with water alone. Field tests showed that sprays containing S. feltiae added to 0.1% Penterra increased WSS mortality up to 29.1%. These results indicate a novel control method for WSS, and represent a significant advancement in the biological control of this persistent insect pest.

Role of podoplanin expression in deciding the invasive potential of ameloblastoma - A retrospective IHC study.

BACKGROUND: The aim of this study was to investigate the podoplanin expression in epithelial odontogenic tumors both non-aggressive and aggressive, tumors with and without ectomesenchyme and remnants of the odontogenic epithelium from dental follicles (DF) of unerupted teeth and to examine its role in progression and invasion of tumors. METHOD: Thirty paraffin embedded specimens AM (15 were non-aggressive ameloblastomas and 15 ameloblastoma showing aggressive behavior), 15 paraffin embedded specimens of AOT, 15 paraffin embedded specimens of CEOT, 15 paraffin embedded specimens of CCOT were obtained and were analyzed by immunohistochemistry using anti-human podoplanin. Podoplanin expression in odontogenic epithelial cells was evaluated using a scoring method, along with calculation of the percentage of positive odontogenic cells. RESULTS: Podoplanin was expressed strongly at the invasive front (in the peripheral odontogenic epithelial cells) of most tumors and dental follicles. Membranous expression of podoplanin in ameloblastomas was stronger in cases of ameloblastomas showing aggressive behavior than (NA) non-aggressive ameloblastomas. CONCLUSION: Expression of podoplanin at the invasive front (in peripheral cells) of odontogenic tumors considered to be associated with neoplastic odontogenic tissues. This molecule might play a role in progression and local invasion of odontogenic tumors. The migration and invasion mediated by podoplanin in odontogenic tumors could be related to cytoskeletal reorganization.

Molecular Characterization Reveals Diverse and Unknown Malaria Vectors in the Western Kenyan Highlands.

The success of mosquito-based malaria control is dependent upon susceptible bionomic traits in local malaria vectors. It is crucial to have accurate and reliable methods to determine mosquito species composition in areas subject to malaria. An unexpectedly diverse set of Anopheles species was collected in the western Kenyan highlands, including unidentified and potentially new species carrying the malaria parasite Plasmodium falciparum. This study identified 2,340 anopheline specimens using both ribosomal DNA internal transcribed spacer region 2 and mitochondrial DNA cytochrome oxidase subunit 1 loci. Seventeen distinct sequence groups were identified. Of these, only eight could be molecularly identified through comparison to published and voucher sequences. Of the unidentified species, four were found to carry P. falciparum by circumsporozoite enzyme-linked immunosorbent assay and polymerase chain reaction, the most abundant of which had infection rates comparable to a primary vector in the area, Anopheles funestus. High-quality adult specimens of these unidentified species could not be matched to museum voucher specimens or conclusively identified using multiple keys, suggesting that they may have not been previously described. These unidentified vectors were captured outdoors. Diverse and unknown species have been incriminated in malaria transmission in the western Kenya highlands using molecular identification of unusual morphological variants of field specimens. This study demonstrates the value of using molecular methods to compliment vector identifications and highlights the need for accurate characterization of mosquito species and their associated behaviors for effective malaria control.

Sex-specific disruption of murine midbrain astrocytic and dopaminergic developmental trajectories following antenatal GC treatment.

The mammalian midbrain dopaminergic systems arising in the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) are critical for coping behaviours and are implicated in neuropsychiatric disorders where early life challenges comprise significant risk factors. Here, we aimed to advance our hypothesis that glucocorticoids (GCs), recognised key players in neurobiological programming, target development within these systems, with a novel focus on the astrocytic population. Mice received antenatal GC treatment (AGT) by including the synthetic GC, dexamethasone, in the mothers' drinking water on gestational days 16-19; controls received normal drinking water. Analyses of regional shapes and volumes of the adult SNc and VTA demonstrated that AGT induced long-term, dose-dependent, structural changes that were accompanied by profound effects on astrocytes (doubling/tripling of numbers and/or density). Additionally, AGT induced long-term changes in the population size and distribution of SNc/VTA dopaminergic neurons, confirming and extending our previous observations made in rats. Furthermore, glial/neuronal structural remodelling was sexually dimorphic and depended on the AGT dose and sub-region of the SNc/VTA. Investigations within the neonatal brain revealed that these long-term organisational effects of AGT depend, at least in part, on targeting perinatal processes that determine astrocyte density and programmed cell death in dopaminergic neurons. Collectively, our characterisation of enduring, AGT-induced, sex-specific cytoarchitectural disturbances suggests novel mechanistic links for the strong association between early environmental challenge (inappropriate exposure to excess GCs) and vulnerability to developing aberrant behaviours in later life, with translational implications for dopamine-associated disorders (such as schizophrenia, ADHD, autism, depression), which typically show a sex bias.

Unexpected diversity of Anopheles species in Eastern Zambia: implications for evaluating vector behavior and interventions using molecular tools.

The understanding of malaria vector species in association with their bionomic traits is vital for targeting malaria interventions and measuring effectiveness. Many entomological studies rely on morphological identification of mosquitoes, limiting recognition to visually distinct species/species groups. Anopheles species assignments based on ribosomal DNA ITS2 and mitochondrial DNA COI were compared to morphological identifications from Luangwa and Nyimba districts in Zambia. The comparison of morphological and molecular identifications determined that interpretations of species compositions, insecticide resistance assays, host preference studies, trap efficacy, and Plasmodium infections were incorrect when using morphological identification alone. Morphological identifications recognized eight Anopheles species while 18 distinct sequence groups or species were identified from molecular analyses. Of these 18, seven could not be identified through comparison to published sequences. Twelve of 18 molecularly identified species (including unidentifiable species and species not thought to be vectors) were found by PCR to carry Plasmodium sporozoites - compared to four of eight morphological species. Up to 15% of morphologically identified Anopheles funestus mosquitoes in insecticide resistance tests were found to be other species molecularly. The comprehension of primary and secondary malaria vectors and bionomic characteristics that impact malaria transmission and intervention effectiveness are fundamental in achieving malaria elimination.

Efficacy of curcumin in the treatment for oral submucous fibrosis - A randomized clinical trial.

INTRODUCTION: Oral submucous fibrosis (OSF) is a chronic, insidious disease that is associated with significant functional morbidity and an increased risk for malignancy. Turmeric and its active ingredient "curcumin" are being studied upon as chemopreventive agents in various diseases. The present study aims to determine the efficacy of curcumin in the treatment of OSF. MATERIALS AND METHODS: Thirty clinically diagnosed OSF patients were divided into two groups, 15 patients in each group from the Outpatient Department. Test group patients were treated with Longvida (curcumin) lozenges and control group with Tenovate ointment (clobetasol propionate (0.05%). The treatment was given for 3 months duration and follow-up was done for 6 months. Both the groups were advised for physiotherapy exercises by mouth exercise device. The baseline and follow-up results were compared for IIO (interincisal distance on maximum mouth opening), Visual Analogue Scale (VAS) for normal food and VAS for spicy food. RESULTS: The test group showed 5.93 (±2.37) mm increase in mouth opening compared to 2.66 (±1.76) mm of the control group. In relation to VAS scale with spicy and normal food the average reduction was 64 (42-73) and 77 (70.5-82) as compared to 34 (14.5-64.5) and 64 (46-75.5) respectively in control group. The test group results achieved in the treatment span was sustained in the follow-up (P < 0.05) compared to control group which showed statistically significant (P < 0.05) relapse. CONCLUSION: It can be concluded that combination strategies for the management of OSF which include the stoppage of causative ill habits, appropriate medicinal and physiotherapy management is more efficient than single therapeutic modality. It is evident from the study that curcumin holds good promise in the treatment of OSF in future.

The role of invertases in plant compensatory responses to simulated herbivory.

BACKGROUND: The ability of a plant to overcome animal-induced damage is referred to as compensation or tolerance and ranges from undercompensation (decreased fitness when damaged) to overcompensation (increased fitness when damaged). Although it is clear that genetic variation for compensation exists among plants, little is known about the specific genetic underpinnings leading to enhanced fitness. Our previous study identified the enzyme GLUCOSE-6-PHOSPHATE DEHYDROGENASE 1 (G6PD1) as a key regulator contributing to the phenomenon of overcompensation via its role in the oxidative pentose phosphate pathway (OPPP). Apart from G6PD1 we also identified an invertase gene which was up-regulated following damage and that potentially integrates with the OPPP. The invertase family of enzymes hydrolyze sucrose to glucose and fructose, whereby the glucose produced is shunted into the OPPP and presumably supports plant regrowth, development, and ultimately compensation. In the current study, we measured the relative expression of 12 invertase genes over the course of plant development in the Arabidopsis thaliana genotypes Columbia-4 and Landsberg erecta, which typically overcompensate and undercompensate, respectively, when damaged. We also compared the compensatory performances of a set of invertase knockout mutants to the Columbia-4 wild type. RESULTS: We report that Columbia-4 significantly up-regulated 9 of 12 invertase genes when damaged relative to when undamaged, and ultimately overcompensated for fruit production. Landsberg erecta, in contrast, down-regulated two invertase genes following damage and suffered reduced fitness. Knockout mutants of two invertase genes both exhibited significant undercompensation for fruit production, exhibiting a complete reversal of the wild type Col-4's overcompensation. CONCLUSION: Collectively, these results confirm that invertases are essential for not only normal plant growth and development, but also plants' abilities to regrow and ultimately compensate for fitness following apical damage.

Estimation of vascular endothelial growth factor gene -460 C/T polymorphism as a biomarker in oral squamous cell carcinoma patients from the Indian subcontinent.

AIM: Oral squamous cell carcinoma (OSCC) is one of the most common malignancies in India. Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis, is known to play a key role in the pathophysiology of tumorigenesis. Polymorphisms of the VEGF gene have been shown to influence the risk of OSCC, but it remains to be elucidated in the Indian population. Thus, the present study was designed to detect the VEGF gene -460 C/T polymorphism in OSCC patients from the Indian population, which is genetically distinct from previously-studied populations. METHODS: Peripheral blood samples of 80 OSCC patients and 80 healthy individuals were collected for a case-control study design. The VEGF gene -460 C/T polymorphism was detected by polymerase chain reaction-based restriction analysis. RESULTS: The analysis revealed significant differences between normal individuals and patients with OSCC (P < 0.05). The distribution of the TT homozygote and the T allele in the patient group was significantly greater than in the control group. CONCLUSION: The present study provides novel evidence of the role of the VEGF gene -460 C/T polymorphism as a suitable genetic marker of OSCC in the Indian population.

Demonstration of mycobacterium tuberculosis in sputum and saliva smears of tuberculosis patients using ziehl neelsen and flurochrome staining- a comparative study.

BACKGROUND: Early detection of tuberculosis is important for reducing its morbidity and mortality especially in the patients with non-productive cough. To overcome the cumbersome process involved in collection and processing of the sputum specimen, the time consumed for reporting of sputum by Ziehl Neelsen (ZN) method and to introduce a routine screening test in suspected, symptomless tuberculosis patients, the present study was designed using saliva as diagnostic medium and Auramine Rhodamine (AR) as staining method. On review of literature, there was no study which has tried diagnosing tuberculosis using saliva with flurochrome stain; hence the present study was designed. AIM: To introduce a routine screening test for tuberculosis patient using saliva and to determine the diagnostic efficacy of routine ZN staining method and AR fluorescent staining method in sputum and saliva smears of pulmonary tuberculosis patients. SETTINGS AND DESIGN: Laboratory settings and Experimental design. MATERIALS AND METHOD: Fifty smears samples of sputum and saliva of known cases of pulmonary tuberculosis were stained with routine ZN stain and other with AR fluorescent stain. All the specimens were inoculated into Lowenstein-Jensen culture media. The smears were subjected for scanning of Mycobacterium tuberculous bacilli under X 1000 magnification for ZN stain and X 400 magnification for AR stain by grid pattern proposed by National tuberculosis institute and graded by RNTCP grading system. RESULTS: All 50 sputum samples showed 100% positivity by ZN and AR stain while only 76% positivity was seen by culture. Of the 50 saliva samples 10% cases were positive by ZN, 76% were positive by AR & 70% by culture method. Statistical analysis using chi square test was done, and the value was found to be statistically highly significant for AR staining technique. (p<0.001) CONCLUSION: Saliva can prove to be an important tool for the diagnosis as well as screening of the patients with pulmonary tuberculosis when aided with flurochrome staining method.