Attenuation of CFA-induced arthritis through regulation of inflammatory cytokines and antioxidant mechanisms by Solanum nigrum L. leaves extracts.

Solanum nigrum L. is a popular traditional medicine for various inflammatory conditions including rheumatism and joint pain. The current study aimed to evaluate the anti-arthritic mechanism of Solanum nigrum L. Four extracts were prepared using n-hexane, methanol, chloroform, and water. The anti-nociceptive and anti-inflammatory activity was carried out with 100, 200, and 300 mg/kg body wt. PO of each extract by the hot plate and carrageenan-induced paw oedema methods, respectively. The anti-arthritic study was performed with chloroform and aqueous extracts (300 mg/kg) in complete Freund's adjuvant (CFA)-induced arthritis. Paw size (mm), ankle joint diameter (mm), and latency time (sec) were recorded on day 0 and every 4th day till 28 days. The hematological, inflammatory, and oxidative biomarkers were estimated. Results showed that significant analgesia (p < 0.05) and reduction in paw inflammation were achieved with all extracts. The highest percent inhibition in Carrageenan-induced inflammation was achieved with 300 mg/kg of chloroform (72.19%) and aqueous (71.30%) extracts, respectively. In the CFA model, both extracts showed a significant reduction in paw size and ankle joint diameter (p < 0.05). The RT-qPCR analysis revealed the upregulation of interleukin-4 and interleukin-10, and down-expression of interleukin-1ß, interleukin-6, tumor necrosis factor-α, cycloxygenase-2, nuclear factor-κB, prostaglandin E synthase 2, and interferon-γ. A significant increase in superoxide dismutase, catalase, and glutathione levels was observed. Hence, it is concluded that Solanum nigrum L. leaf extracts regulate the expression of inflammatory markers and improve oxidative stress resulting in the attenuation of CFA-induced arthritis.

Assessment of in vivo antiepileptic potential and phytochemical analysis of Cassia absus seed extracts.

Cassia absus, a member of Fabaceae family, has been a part of traditional medicine for various ailments such as Hypertension, Diabetes, and Cancer. This family of plants has been utilized for Anticonvulsant and Anxiolytic effects. The ongoing investigation is aimed to seek the antiepileptic potential of C. absus seed extracts in pentylenetetrazole-induced kindling mice. The seeds of C. absus were subjected to a sequential extraction process for the preparation of n-hexane, chloroform, methanol, and aqueous extracts. The PTZ-induced kindling model was employed to assess the antiepileptic activity of each extract. Seizure activity and antioxidant biomarkers in the brain tissue such as levels of CAT, SOD, tGSH, and MDA were assessed. Mechanism of action was elucidated by Flumazenil. Through GC-MS analysis, the phytochemical components in the chloroform extract of C. absus were evaluated. The outcomes showed that C. absus extracts markedly reduced the seizure activity in kindling mice. The extracts exhibited significant Antioxidant properties by enhancing the levels of antioxidant biomarkers in the brain tissue such as CAT, SOD, and tGSH, and decreasing the MDA level. The results demonstrated that C. absus extracts showed antiepileptic effects may be via GABA pathway. According to the results of this investigation, C. absus has significant antiepileptic potential in PTZ-induced kindling mice via GABA pathway modulation and combating reactive oxygen species.

Cassia absus-mediated upregulation of IL-4, IL-10 and downregulation of IL-1ß, IL-6, TNF- α, NF-κB, IFN-γ in CFA-induced arthritis model.

Traditional use of Cassia absus as an anti-inflammatory in conjunctivitis and bronchitis is well reported. Owing to its anti-inflammatory potential, the current study appraised in vivo anti-arthritic activity of n-hexane and aqueous extracts of Cassia absus seeds (200 mg/kg) using Complete Freund's Adjuvant (CFA) rat model of arthritis. Changes in paw size (mm), joint diameter (mm), and pain response (sec) were recorded at the baseline and then after CFA induction at the interval of 4 days till the 28th day. Blood samples of anesthetized rats were collected for the estimation of hematological, oxidative, and inflammatory biomarkers. Results showed percent inhibition in paw edema (45.09% and 60.79%) with both n-hexane and aqueous extracts, respectively. Significant reduction in paw size and ankle joint diameter (P < 0.01) was seen in extracts treated rats. Erythrocyte Sedimentation rate, C-Reactive Protein, White Blood Cell levels significantly lowered, and Hemoglobin, Platelets and Red Blood Cell count significantly increased post-treatments. Superoxide Dismutase, Catalase, and Glutathione were significantly improved (P < 0.0001) in treated groups as compared to CFA induced arthritic control. Real-time polymerase chain reaction investigation showed significant downregulation (P < 0.05) of Interleukin-1ß, Tumor Necrosis Factor-α, Interleukin-6, Cycloxygenase-2, Nuclear Factor-κB, Prostaglandin E Synthase 2, Interferon Gamma and upregulation of Interleukin-4, Interleukin-10 in both n-hexane and aqueous extract-treated groups. It is thereby concluded that Cassia absus can significantly attenuate CFA-induced arthritis by modulation of oxidative and inflammatory biomarkers.

Biomolecular Screening of Pimpinella anisum L. for Antioxidant and Anticholinesterase Activity in Mice Brain.

Hundreds of the plants have been explored and evaluated for antioxidant and anti-amnesic activities, so far. This study was designed to report the biomolecules of Pimpinella anisum L. for the said activities. The aqueous extract of dried P. anisum seeds was fractionated via column chromatography and the fractions so obtained were assessed for the inhibition of acetylcholinesterase (AChE) via in vitro analysis. The fraction which best inhibited AChE was so named as the P. anisum active fraction (P.aAF). The P.aAF was then chemically analyzed via GCMS, which indicated that oxadiazole compounds were present in it. The P.aAF was then administered to albino mice to conduct the in vivo (behavioral and biochemical) studies. The results of the behavioral studies indicated the significant (p < 0.001) increase in inflexion ratio, by the number of hole-pokings through holes and time spent in a dark area by P.aAF treated mice. Biochemical studies demonstrated that the oxadiazole present in P.aAF on one hand presented a noteworthy reduction in MDA and the AChE level and on the other hand promoted the levels of CAT, SOD and GSH in mice brain. The LD50 for P.aAF was calculated as 95 mg/Kg/p.o. The findings thus supported that the antioxidant and anticholinesterase activities of P. anisum are due to its oxadiazole compounds.

Metabolic Profiling and Investigation of the Modulatory Effect of Fagonia cretica L. Aerial Parts on Hepatic CYP3A4 and UGT2B7 Enzymes in Streptozotocin-Induced Diabetic Model.

Drug-metabolizing enzymes are either boosted or suppressed by diabetes mellitus. This research was designed to explore Fagonia cretica L. aerial parts' impact on CYP3A4 and UGT2B7 activity and their mRNA expression in diabetic rats. Fagonia cretica (F. cretica) dried powder was sequentially extracted with n-hexane, chloroform, ethyl acetate, methanol, and water. The methanol extract and aqueous fraction presented the most significant potential to decrease the concentration of alpha-hydroxyl midazolam, with 176.0 ± 0.85 mg/Kg and 182.9 ± 0.99 mg/Kg, respectively, compared to the streptozotocin (STZ)-induced diabetic group, reflecting the inhibition in CYP3A4 activity. The fold change in mRNA expression of CYP3A4 was decreased significantly by the methanol extract, and the aqueous fraction of F. cretica estimated by 0.15 ± 0.002 and 0.16 ± 0.001, respectively, compared with the diabetic group. Morphine metabolism was significantly increased in rats treated with F. cretica methanol extract and its aqueous fraction, displaying 93.4 ± 0.96 mg/Kg and 96.4 ± 1.27 mg/Kg, respectively, compared with the metabolism of morphine in the diabetic group, which highlights the induction of UGT2B7 activity. The fold change in mRNA expression of UGT2B7 was significantly increased by the methanol extract and the aqueous fraction, estimated at 8.14 ± 0.26 and 7.17 ± 0.23 respectively, compared to the diabetic group. Phytochemical analysis was performed using high-performance liquid chromatography (HPLC), where the methanol extract showed more flavonoids and phenolic compounds compared to the aqueous fraction of F. cretica. The obtained results were further consolidated by molecular docking studies, where quercetin showed the best fitting within the active pocket of CYP3A4, followed by gallic acid, displaying free binding energies (∆G) of -30.83 and -23.12 kcal/mol, respectively. Thus, F. cretica could serve as a complementary medicine with standard anti-diabetic therapy that can modulate the activity of the drug-metabolizing enzymes.

Evaluation of the Anxiolytic and Anti-Epileptogenic Potential of Lactuca Serriola Seed Using Pentylenetetrazol-Induced Kindling in Mice and Metabolic Profiling of Its Bioactive Extract.

This study aimed to assess the potential of Lactuca serriola (Asteraceae) seed n-hexane, chloroform, methanol, and aqueous extracts as anticonvulsant, sedative, anticonvulsant and antiepileptic agents in Swiss albino mice. Different doses of each extract were evaluated for the anxiolytic potential using the hole-board, the elevated plus maze and the light/dark test. A phenobarbitone-induced sleep test was employed for the evaluation of sedative potential. Acute anticonvulsant activity was evaluated by picrotoxin and strychnine-induced convulsion models. All extracts significantly reduced the number of head dips where n-hexane extract (400 mg/kg) showed 96.34% reduction in the tendency of head dipping when compared with the control. Mice treated with extracts preferred elevated plus maze open arms and were shown to lack open arms evasion, especially n-hexane extract (400 mg/kg)-which showed 456.14%-increased the duration of open arm stay with the respective control group. By reducing sleep latency and greatly lengthening sleep duration, L. serriola enhanced the effects of barbiturate-induced sleep. A significant increase in convulsion latency and decrease in convulsions induced by picrotoxin and strychnine duration was observed in all extract-treated groups. All the extracts exhibited anti-epileptogenic potential as the seizure score in pentylenetetrazol (PTZ)-induced kindling in mice was reduced significantly. Maximum protection was afforded by chloroform extract that reduced the seizure score by 79.93% compared with the PTZ group. Chloroform executed antioxidant effect by elevating super oxide dismutase (SOD) by 126%, catalase (CAT) by 83.53%, total glutathione (tGSH) by 149%, and reducing malondialdhyde (MDA) levels by 36.49% in the brain tissues that is further consolidated by histopathological examination. Metabolic profiling of the most active chloroform extract using Gas chromatography coupled with mass showed the presence of 16 compounds. This anti-epileptic activity was further confirmed via in silico molecular modelling studies in the active site Gamma-aminobutyric acid aminotransferase (GABA-AT) where all of the tested metabolites illustrated a potent inhibitory potential towards GABA-AT with hexadecanoic acid, 15-methyl-, methyl ester followed by octadecanoic acid, methyl ester showed the best fitting. The results indicated the possible anxiolytic and anti-epileptogenic potential of the plant and further consolidated the ethnopharmacological use of L. serriola seeds.

Anti-arthritic, anti-nociceptive and anti-inflammatory potential of Cassia absus: An ethanomedicinal plant of Febaceae family.

Medicinal plants are becoming popular choice for treatment of chronic disease conditions. Cassia absus plant parts have been traditionally used to treat inflammatory conditions. This study was designed for evaluating anti-arthritic, anti-nociceptive and anti-inflammatory potential of Cassia absus seeds. n- hexane, methanol, chloroform and aqueous extracts were prepared to be appraised for identification and quantitative determination of various phytochemicals. All the extracts were evaluated for anti-arthritic activity via protein denaturation, anti-nociceptive activity by hot plate method and anti-inflammatory potential through Carrageenan induced paw edema. Three doses; 100, 200 and 300mg/kg of each extract were given to Wistar rats. The results of the quantitative analysis revealed that Aqueous and n-hexane extracts contained the highest total flavonoid (104.2±0.24mg QE/g) and phenolic contents (187.4±0.65mg GA/g) respectively. All the extracts exhibited decrease in protein denaturation (n-hexane 66.66%, methanol 59.42%, chloroform 65.21% and aqueous extract 89.85%). Significant increase in mean latency time (secs) was observed in n-hexane, methanol and aqueous extract treated rats as compared to normal rats. All four extracts caused significant reduction in paw inflammation as compared to carrageenan control. It is therefore concluded that all the extracts of Cassia absus possessed significant anti-arthritic, anti-nociceptive and anti-inflammatory potential.

Evaluation of phytochemical, anti-oxidant and cardiac depressant effect of Rumex dentatus by using Langendorff's isolated heart apparatus.

Rumex dentatus has been used traditionally for ailment of cardiovascular diseases. The aim of the present study was to assess cardiovascular effects in isolated perfused rabbit heart. Aqueous and n-butanolic fractions were assessed for their effect on perfusion pressure (PP), force of contraction (FC) and heart rate (HR) of rabbit heart using Langendorff's method. The possible mechanisms of action of extracts/fraction were assessed with and without application of different agonist/antagonist. Phytochemical, toxicity and anti-oxidant activities were also determined. Both extracts at 1mg/mL dose produced a highly significant decrease in FC and HR but PP remained unchanged. Moreover, aqueous fraction of Rumex dentatus at 0.001mg/mL dose produced a highly significant decrease in FC and HR but no significant change in PP was observed. Atropine 10-5 M did not inhibit the cardiac depressant response of both fractions. Furthermore, both fractions blocked the positive ionotropic and chronotropic effects of adrenaline and calcium chloride. Phytochemical studies have shown the presence of some phytochemicals. Acute and sub-chronic toxicity studies demonstrated that test extracts are safe and produced no significant changes in haematological and biochemical parameters. Crude extract showed significant antioxidant activity like ascorbic acid. This study revealed that this plant have good cardiac depressant effect.

Biomolecular Evaluation of Lavandula stoechas L. for Nootropic Activity.

Lavandula Stoechas L. is widely known for its pharmacological properties. This study was performed to identify its biomolecules, which are responsible for enhancement of memory. L. stoechas aqueous extract was first purified by liquid column chromatography. The purified fractions were analyzed for in vitro anti-cholinesterase activity. The fraction that produced the best anti-cholinesterase activity was named an active fraction of L. stoechas (AfL.s). This was then subjected to GC-MS for identifications of biomolecules present in it. GC-MS indicated the presence of phenethylamine and α-tocopherol in AfL.s. Different doses of AfL.s were orally administered (for seven days) to scopolamine-induced hyper-amnesic albino mice and then behavioral studies were performed on mice for two days. After that, animals were sacrificed and their brains were isolated to perform the biochemical assay. Results of behavioral studies indicated that AfL.s improved the inflexion ratio in mice, which indicated improvement in retention behavior. Similarly, AfL.s significantly (p < 0.001) reduced acetylcholinesterase and malondialdehyde contents of mice brain, but on the other hand, it improved the level of choline acetyltransferase, catalase, superoxide dismutase, and glutathione. It was found that that high doses of AfL.s (≥400 mg/Kg/p.o.) produced hyper-activity, hyperstimulation, ataxia, seizures, and ultimate death in mice. Its LD50 was calculated as 325 mg/Kg/p.o. The study concludes that α-tocopherol and phenethylamine (a primary amine) present in L. stoechas enhance memory in animal models.

Appraisal of the Antiarthritic Potential of Prazosin via Inhibition of Proinflammatory Cytokine TNF-α: A Key Player in Rheumatoid Arthritis.

Prazosin, a selective α1 adrenergic receptor antagonist, with documented anti-inflammatory potential, was evaluated for its antiarthritic efficacy by targeting specifically TNF-α. The antiarthritic attribute of prazosin validated through in vitro screening comprised thermally provoked denaturation of bovine serum albumin (BSA) and egg albumin along with membrane stabilization evaluation at a concentration of 100-6400 µg/mL, while in vivo screening comprised formaldehyde-instigated arthritis at the doses of 5, 10, and 20 mg/kg and complete Freund's adjuvant (CFA)-induced arthritis at 20 mg/kg dose. Paw swelling, body weight, arthritic score, hematological parameters, and histological and radiographic examination of ankle joints were assessed for a period of 28 days after CFA immunization. Moreover, the proinflammatory cytokine TNF-α level was also assessed through quantitative real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Prazosin revealed significant antiarthritic effect evident through protein denaturation inhibition in the egg albumin and the BSA model, stabilization of red blood cell membrane in the membrane stabilizing assay, and reduction in paw volume in formaldehyde-induced arthritis. Likewise, prazosin exhibited propitious antiarthritic effects in the CFA-provoked arthritis model manifested by paw volume and arthritic score alleviation, substantial weight loss prevention, and preservation of the normal hematological and biochemical profile. Histological and X-ray investigation unveiled no substantive structural alterations in treated rat's ankle joints. The TNF-α expression level was also reduced. Thus, the current study is suggestive that prazosin exhibits a strong antiarthritic potential possibly through inhibition of TNF-α.

Amelioration of Bleomycin and Methotrexate-Induced Pulmonary Toxicity by Serratiopeptidase and Fisetin.

Pulmonary toxicity by anticancer drugs often leads to discontinuation of therapy or switching the therapy to alternative drugs. In the present study, serratiopeptidase (SPTD) and fisetin (FST) were evaluated as chemoprotectant to counteract the pulmonary toxicity induced by BLM and MTX. Single dose of MTX (20 mg/kg) by intraperitoneal and BLM (5 mg/kg) by intra-tracheal route was administered on 7th day of study. SPTD (20 mg/kg), FST (25 mg/kg), and NAC (250 mg/kg) and combinations of SPTD + NAC, SPTD + FST, and FST + NAC were administered through oral gavage for 14 days. SPTD and FST showed significant (p < 0.05) effect in MTX-induced lung toxicity by increasing reduced glutathione (GSH) and decreasing malondialdehyde (MDA), hydroxyproline (HXP), and collagen. SPTD and NAC showed significant (p < 0.05) effect in BLM-induced pulmonary toxicity by increasing GSH and decreasing MDA, HXP, and collagen whereas FST was not much effective. In combination study, SPTD + NAC combination showed significant (p < 0.05) effect in BLM- and MTX- induced lung injury whereas other combinations did not prove to be highly effective. SPTD can be recommended along with BLM and MTX in chemotherapy protocol alone and in combination with NAC.

Green synthesis of gold nanoparticles from Manilkara zapota L. extract and the evaluation of its intrinsic in vivo antiarthritic potential.

The plant Manilkara zapota belongs to the family Sapotaceae and is commonly known as Chiku in Pakistan. Traditionally, M. zapota is used in pulmonary diseases, diarrhea, rheumatism, hemorrhage, and ulcers. There is no study available on the in vivo antiarthritic activity of M. zapota and its gold nanoparticles (AuNPs). The aim of this study is to evaluate the in vivo acute and sub-acute antiarthritic activity of aqueous extract of M. zapota and its biosynthesized gold nanoparticles (AuNPs). Plant-induced reduction method was used for the synthesis of AuNPs. The synthesized AuNPs were characterized via UV, FTIR, SEM, and zeta potential measurements and were optimized by screening various parameters including time, temperature, pH, and salt concentration. Arthritis in rats was induced by Freund's Complete Adjuvant (FCA) injection in hind paw. The antiarthritic effect was evaluated by the determination of paw volume, joint diameter, latency time, hematological, biochemical parameters, antioxidant biomarkers, TNF-α level, and radiological evaluation. The aqueous extract and nanoparticles significantly decreased the paw volume, joint diameter, and significantly increased latency time as compared to the FCA-induced arthritic group. They significantly normalized the hematological, biochemical parameters, and oxidative stress biomarkers in comparison to the arthritic group. They also significantly decreased the TNF-α level when assessed against the arthritic group. Radiological evaluation confirmed the antiarthritic effect of the aqueous extract and nanoparticles of M. zapota leaf extract. It is concluded that the aqueous extract and nanoparticles of M. zapota possess significant analgesic, antiarthritic, and anti-inflammatory activity. However, nanoparticles possess more pronounced antiarthritic activity as compared to the aqueous extract. Moreover, free radical scavenging action and TNF-α reduction showed a prominent role in their antiarthritic activity. Further, investigation is underway to identify the active phytochemical constituent responsible for the antiarthritic activity.

Lavandula stoechas (L) a Very Potent Antioxidant Attenuates Dementia in Scopolamine Induced Memory Deficit Mice.

The objective of the current project was to explore the pharmacotherapeutic role of Lavandula stoechas (L) for the management of dementia. Dementia is considered a global challenge of current century seeking special attention of pharmacologists to explore its best remedies. Methanolic extract of aerial parts of L. stoechas was tested for phytochemical analysis along with free radical scavenging activity. Behavioral studies were performed on scopolamine induced amnesic mice by using elevated plus maze (EPM), light and dark test and hole board paradigms. Biochemical investigations were made after decapitating the mice. Their brains were isolated for biochemical estimation of acetylcholinesterase (AChE), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). Phytochemical study ensured the presence of total phenolic contents (285.91 ± 0.75 mg of GAE/g of extract), total flavonoids (134.06 ± 0.63 mg of RE/g of extract), total tannins (149.60 ± 0.93 mg of TAE/g of extract) and free radical scavenging activity (IC50 value = 76.73 µg/ml found by DPPH method). Behavioral studies indicated that animals of GVII showed higher inflexion ratio (0.40 ± 0.03) for EPM, spent most of time (227.17 ± 2.13 s) in dark area of light dark test and had many hole pockings (39.83 ± 1.88) for hole board paradigm. Moreover, biochemical studies revealed that methanolic extract of L. stoechas (800 mg/kg/p.o.) significantly (P < 0.001) reduced brain AChE and MDA levels while improved SOD, CAT, and GSH levels. Thus the findings suggest that L. stoechas stabilizes memory by enhancing cholinergic neurotransmission and by providing defense against oxidative stress in mice brain.

Glutathione sulfotransferase inhibition activity of a self-fermented beverage, Kanji.

CONTEXT: Kanji, a liquid preparation of roots of Daucus carota L. ssp. sativus (Hoffm.) Arcang. var. vavilovii Mazk. (Apiaceae), may inhibit glutathione sulfotransferase (GST) activity due to ferulic acid content. OBJECTIVES: GST inhibition activity and characterization of Kanji and methanol extract of D. carota roots, and oral absorption pattern of ferulic acid from Kanji in rats. MATERIALS AND METHODS: GST inhibition activity of Kanji and methanol extract of D. carota roots in concentration range 0.001-100.00 mg/mL was determined using Sprague Dawley rat liver cytosolic fraction. Methanol extract upon column chromatography gave ferulic acid, which was used to characterize Kanji and determine its oral absorption pattern in Wistar rats. RESULTS: The GST inhibition activity of Kanji (100.00 µg/mL), methanol extract of D. carota roots (100.00 µg/mL) and tannic acid (10.00 µg/mL, positive control) was found to be 0.162 ± 0.016, 0.106 ± 0.013 and 0.073 ± 0.004 µM/min/mg, respectively. Different Kanji samples and methanol extract contained ferulic acid (0.222-0.316 mg/g) and 0.77 mg/g, respectively. Ferulic acid did not appear in plasma after oral administration of Kanji. DISCUSSION: Kanji having solid contents 80.0 µg/mL, equivalent to 0.0025 µg/mL ferulic acid, does not inhibit the activity of GST. The oral administration of Kanji, in human equivalent dose (528 mg/kg, 16.67 µg ferulic acid), to rats indicated poor absorption of ferulic acid. CONCLUSION: Kanji having solid contents 14-36 mg/mL does not inhibit GST activity, hence may not interfere with drugs that are the substrates of GST, if taken concomitantly.

Risk factors for multi-drug resistant tuberculosis in patients at tertiary care hospital, Peshawar.

OBJECTIVE: To determine the frequency of common risk factors for multi-drug resistant tuberculosis in patients presented at a tertiary care hospital, Peshawar. STUDY DESIGN: Cross-sectional, observational study. PLACE AND DURATION OF STUDY: Pulmonology Department, Khyber Teaching Hospital, Peshawar, from December 2006 to October 2007. METHODOLOGY: Patients with positive AFB culture and sensitivity results and found resistant to both rifampicin and isoniazid with or with resistance to other first line anti-tuberculosis drugs, were recruited from both Pulmonology Ward and Outpatient Department (OPD). Informed verbal consent was taken and a questionnaire administered to all participants of the study. Information regarding demographics, education status, occupation, monthly household income, AFB C/S, details of past history of tuberculosis and family history of TB or Multi-Drug Resistant (MDR-TB) was recorded. Data was analyzed on SPSS version 11. RESULTS: A total of 30 patients of MDR-TB were interviewed. Male (n=17) and female (n=13) ratio was 1.3:1. Mean age was 34.2+/-15.3 years. Ninety-two percent female and 52.9% male were uneducated. In 56.7%, monthly income was less than 5,000 rupees and in 40% between 5,000-10,000 rupees. All patients had previous history of Antituberculous Treatment (ATT), in which 20% had undertaken ATT course once, 53.3% twice and 26.7% thrice in the past. In the study group, 13 (43.3%) patients had not completed their first ATT course and 11 of them were receiving ATT from a general practitioner (GP) at that time. Seven (23.3%) patients had family history of TB but no one had documented MDR-TB in the family. Resistance to RH was present in all patients; moreover, 56.7% had resistance to RHEZ+S. CONCLUSION: The most common factors in the study group were previous history of tuberculosis, repeated courses of ATT, prescribed by different clinicians and unsupervised treatment by a GP during the initial course of ATT.