Sialic acid blockade in dendritic cells enhances CD8+ T cell responses by facilitating high-avidity interactions.

Sialic acids are negatively charged carbohydrates that cap the glycans of glycoproteins and glycolipids. Sialic acids are involved in various biological processes including cell-cell adhesion and immune recognition. In dendritic cells (DCs), the major antigen-presenting cells of the immune system, sialic acids emerge as important regulators of maturation and interaction with other lymphocytes including T cells. Many aspects of how sialic acids regulate DC functions are not well understood and tools and model systems to address these are limited. Here, we have established cultures of murine bone marrow-derived DCs (BMDCs) that lack sialic acid expression using a sialic acid-blocking mimetic Ac53FaxNeu5Ac. Ac53FaxNeu5Ac treatment potentiated BMDC activation via toll-like receptor (TLR) stimulation without affecting differentiation and viability. Sialic acid blockade further increased the capacity of BMDCs to induce antigen-specific CD8+ T cell proliferation. Transcriptome-wide gene expression analysis revealed that sialic acid mimetic treatment of BMDCs induces differential expression of genes involved in T cell activation, cell-adhesion, and cell-cell interactions. Subsequent cell clustering assays and single cell avidity measurements demonstrated that BMDCs with reduced sialylation form higher avidity interactions with CD8+ T cells. This increased avidity was detectable in the absence of antigens, but was especially pronounced in antigen-dependent interactions. Together, our data show that sialic acid blockade in BMDCs ameliorates maturation and enhances both cognate T cell receptor-MHC-dependent and independent T cell interactions that allow for more robust CD8+ T cell responses.

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.

Nickel, its adverse health effects & oxidative stress.

Nickel-induced toxicity and carcinogenicity, with an emphasis on the generation and role of reactive oxygen species is reviewed. Nickel is a known haematotoxic, immunotoxic, neurotoxic, genotoxic, reproductive toxic, pulmonary toxic, nephrotoxic , hepatotoxic and carcinogenic agent. This article presents a selective review on nickel and effect of its acute, subchronic and chronic doses on certain metabolically active tissues in human as well as animals. Nickel exposure causes formation of free radicals in various tissues in both human and animals which lead to various modifications to DNA bases, enhanced lipid peroxidation, and altered calcium and sulphydryl homeostasis. The primary route for nickel toxicity is depletion of glutathione and bonding to sulphydryl groups of proteins. Nickel homeostasis, nickel-induced activation of signaling pathways and the protective role of enzymatic and non-enzymatic antioxidants against nickel toxicity and carcinogenicity are also discussed.

Characteristics of patients admitted with stroke.

Population based study on stroke morbidity and mortality is lacking in our country. We described the clinical pattern of patients with stroke admitted in Neuromedicine Unit, Chittagong Medical College Hospital, Bangladesh. One hundred and six consecutive patients were included in the study. Clinical diagnosis of stroke, initial assessment, and assessment of outcome of stroke were performed by the neurologists. On admission blood glucose, and creatinine level was estimated, and an electrocardiogram was recorded. Fasting lipid level estimation and CT-scan of brain were performed for patients who could afford the cost. The mean age of the patients was 60.0 +/-13.7 years and the highest occurrence of stroke was found in the age group of 61 - 70 years. The proportions of rural, semiurban and urban patients were 46.2%, 27.4% and 26.4% respectively. A large portion of the patients were found illiterate (47.2%), and only one patient had postgraduate education. The average per capita income was found 1159 +/-762 taka per month. About one-fourth of the patients had diabetes (21.7%). Hypertension, ischemic heart disease and dyslipidemia were found in 59.4%, 18.9% and 11.3% respectively. The cause of hospitalization was altered consciousness (58.5%), right hemiplegia (32.1%), and left hemiplegia (31.1%) either alone or in combination. The mean duration of hospital stay was 5.25 +/-2.19 days. Fifty percent partially and 35% satisfactorily recovered, and 7 (6.6%) patients expired. Patients from lower socioeconomic group were admitted in the hospital. A considerable number of stroke patients had hypertension and diabetes.

Rational Hyperoxia in the Perioperative Period: a Safe and Effective Tool in the Reduction of SSI.

Oxygen supplemented at a concentration higher than 40-50 % for at least 2 h perioperatively is expected to reduce surgical site infections (SSI). Although supplementation of 80 % of oxygen perioperatively has shown to reduce SSI in various studies, this concentration is known to be associated with airway complications. This study was taken up to assess the efficacy of 60 %, i.e. <80 and >50 %, inspired oxygen supplemented perioperatively in reducing SSI. One hundred and eighty-eight patients who underwent elective class I and II surgeries were studied. Patients were divided equally into two groups and subgroups and matched for age, sex, type of surgeries, etc. The control group received 30 % and the study group received 60 % oxygen supplementation perioperatively for 2 h. Wounds were observed for the development of SSI. 8/94 patients in the study group and 13/94 patients in the control group developed SSI (p < 0.01). The results indicate a relative risk of 1.62, risk difference of 0.0531 and attributable risk of 38.42 %. Hence, it may be concluded that perioperative oxygen supplementation at 60 % concentration reduces SSI.

Solubilized glycosyltransferases and biosynthesis in vitro of glycolipids.

The assembly of most of the ceramide-linked glycolipids (GSLs) in eukaryotic cells occurs in Golgi bodies. At least 18 different glycolipid:glycosyltransferases (GSL:GLTs) have been characterized, 10 of which have been solubilized. These GLTs can be classified into 2 distinct groups: 1) GLTs dedicated to either Dol-P-P-sugar(s) or ceramide-linked sugar(s); and 2) GLTs with dual loyalties (i.e., they compete with glycolipid- and glycoprotein-bound oligosaccharides). Studies with solubilized and purified GalNAcT-1 and GalNAcT-2 from embryonic chicken brains prove that GalNAcT-1 (UDP-GalNAc:GM3 beta 1-4GalNAcT) is specific for GSL, whereas GalNAcT-2 (UDP-GalNAc:Gb3 beta 1-3GalNAcT) can transfer to an oligosaccharide containing the alpha-linked terminal galactose. Similarly, GalT-3 (UDP-Gal:GM2 beta 1-3GalT) is more specific for ganglio-oligosaccharide and GalT-4 (UDP-Gal:Lc3 beta 1-4GalT) can transfer galactose to N-acetylglucosamine linked to p-nitrophenol, glycolipid or glycoprotein. Both GalT-3 and GalT-4 have been separated and purified from embryonic chicken brains. Studies with solubilized SAT-4 and SAT-3, from bovine spleen and embryonic chicken brains, respectively, suggest the existence of 2 different gene-expressed alpha 2-3SATs. The newly discovered FucT-3 (GDP-Fuc:NeuGc-iLc6-alpha 1-3FucT) from human colon carcinoma (Colo-205) has also been solubilized and separated from other GSL:GLTs. Using a new activity gel-Western blot combined technique, the molecular mass of this FucT-3 was determined to be 105 kDa.

Efficacy of ciprofloxacin in enteric fever: comparison of treatment duration in sensitive and multidrug-resistant Salmonella.

The efficacy of two regimens of ciprofloxacin was compared in a randomized study conducted on 69 patients with enteric fever, 52.2% of whom had infection with multidrug-resistant (MDR) strains of Salmonella typhi or S. paratyphi. Patients were randomly assigned to two regimens (10 days versus 14 days) of ciprofloxacin (500 mg twice a day). The mean +/- SD time required for defervescence was similar for both regimens (4.2 +/- 1.9 days in the 10-day group and 4.9 +/- 2.6 days in the 14-day group). A 100% cure was observed in each treatment group and no serious side effects were observed. Relapse occurred in two patients (14-day regimen). Only one patient (14-day regimen) had growth of S. typhi in stool culture at the time of the first follow-up three days after completion of therapy. Follow-up studies on available patients on two, six, and 12 months after completion of therapy revealed that all patients had negative stool cultures for S. typhi and S. paratyphi. This study indicates that ciprofloxacin may be recommended as an initial therapy for enteric fever for adult men and nonpregnant and nonlactating women in areas where MDR strains of S. typhi and S. paratyphi are prevalent, and that 500 mg twice a day of the drug given for 10 days is as effective as 14 days at the same dosage.

In vitro biosynthesis of GbOse4Cer (globoside) and GM2 ganglioside by the (1-->3) and (1-->4)-N-acetyl beta-D-galactosaminyltransferases from embryonic chicken brain. Solubilization, purification, and characterization of the transferases.

(1-->4)-N-Acetyl-beta-D-galactosaminyltransferase (GalNAcT-1) and (1-->3)-N-acetyl-beta-D-galactosaminyltransferase (GalNAcT-2), which are involved in the in vitro biosynthesis of GM2 and GbOse4Cer glycosphingolipids, respectively, have been solubilized and separated by differential detergent extraction from a membrane preparation of 19-day-old embryonic chicken brain. The separated GalNAcT-1 activity had a pH optima of 7.8-8.0, and the separated GalNAcT-2 activity a single pH optimum of 7.2. Furthermore, the partially purified GalNAcT-2 preparation catalyzed the transfer of N-acetylgalactosamine from UDP-D-[3H]GalNAc to only GbOse3Cer and nLcOse5Cer. Both GalNAcT-1 and GalNAcT-2 activities were purified to approximately 316- and 428-fold, respectively, by use of UDP-hexanolamine-Sepharose 4B affinity-column chromatography. However, the partially purified GalNAcT-1 preparation appeared to be active only with GM3, lactosylceramide, and lactotriaosylceramide. The proposed linkage of the N-acetylgalactosamine unit incorporated into GM3 is beta-D-GalpNAc-(1-->4)-GM3 from the isolation of [3H]threitol after hydrolysis of the desialylated, lead tetraacetate-treated, enzymic product, beta-D-GalpNAc-(1-->4)-beta-D-[6-3H]Galp-(1-->4)-beta-D-Glcp-(1-->1)-Cer . In addition, beta-D-GalpNAc-(1-->3)-GbOse3Cer was produced, as shown by the identification of 2,4,6-tri-O-methyl-galactose after permethylation and hydrolysis of the GalNAcT-2 enzymic product, GalpNAc-[6-3H]Galp--->Gal-->Glc-->Cer.

Biosynthesis in vitro of a globoside containing a 2-acetamido-2-deoxy-beta-D-galactopyranosyl group (1----3)-linked and Forssman glycolipid by two N-acetylgalactosaminyltransferases from chemically transformed guinea pig cells.

Two N-acetylgalactosaminyltransferase activities (GalNAcT-2 and GalNAcT-3) have been characterized in chemically transformed, cultured guinea-pig cell lines (104C1 and 106B). Line 104C1 is a benz[a]pyrene-transformed tumorigenic variant, whereas line 106B is a 7,12-dimethylbenz[a]anthracene-transformed nontumorigenic variant obtained from fetal guinea-pig cells at 43 days of gestation. The GalNAcT-2 (UDP-GalNAc:GbOse3Cer beta-N-acetylgalactosaminyltransferase) isolated from both 104C1 and 106B cells catalyzed the transfer of Gal-NAc from UDP-GalNAc to the 3H-labeled terminal galactose group of Gb3 [( 6-3H]Gal alpha 1----4Gal beta 1----4Glc----Cer). The 3H-labeled globoside was purified and then subjected to exhaustive methylation. After acetolysis, the partially methylated sugars were separated by two-dimensional, thin-layer chromatography. 3H-Label was detected in two major areas, 2,4,6-tri-O-Me-Gal (40%) and 2,3,4,6-tetra-O-Me-Gal (46%). In a separate experiment, 80% of the GalNAc was released when labeled GbOse4Cer [( 3H]GalNAc----Gal alpha 1----4Gal beta 1----4Glc----Cer) was treated with purified clam beta-hexosaminidase. The present results establish the formation of a beta-D-GalpNAc-(1----3) linkage in the terminal region of the biosynthesized globoside. GalNAcT-3 activity (UDP-GalNAc:GbOse4Cer alpha-GalNAc-transferase), which catalyzes the transfer of GalNAc from UDP-[14C]- or -[3H]GalNAc to GbOse4Cer (GalNAc beta 1----3Gal alpha 1----4Gal beta 1----4Glc----Cer), was three times higher in 106B cells than in 104C1 cells. The isolated, purified radioactive product formed an immunoprecipitin line against rabbit anti-Forssman antibody.

Biosynthesis in vitro of core lacto-series glycosphingolipids by N-acetyl-D-glucosaminyltransferases from human colon carcinoma cells, Colo 205.

Two N-acetyl-D-glucosaminyltransferases have been detected in human colon carcinoma Colo 205 cells. These enzymes catalyze the biosynthesis in vitro of the core-glycolipid of Type 1 and Type 2 lacto-series antigens and of the polylactosamine-containing longer chain antigenic structures, respectively. The first enzyme, GlcNAcT-1, which catalyzes the formation of lactotriosylceramide [LcOse3Cer, beta-D-GlcpNAc-(1----3)-LcOse2Cer, the core for all lacto-series Type 1 and Type 2 chains] from lactosylceramide [beta-D-Galp-(1----4)-D-Glcp-Cer, LcOse2Cer] and UDP-GlcNAc shows optimum activity in the presence of nonionic detergent Triton CF-54. The other enzyme, GlcNAcT-2, which catalyzes the biosynthesis in vitro of iLcOse5Cer [beta-D-GlcpNAc-(1----3)-nLcOse4Cer, the core for polylactosamine-containing antigens] from nLcOse4Cer [beta-D-Galp-(1----4)-LcOse3Cer] and UDP-GlcNAc, is optimally active with the zwitterionic detergent, Zwittergent 3-14, when membrane-bound. Both of these activities, however, can be extracted from the membrane by use of a nonionic detergent. Triton X-114, with nearly the same efficiency. These two transferases showed different pH optima, different cation and anion effects, and differential heat-inactivation patterns at 55 degrees. Permethylation studies of the radioactive products isolated from both of the enzyme-catalyzed reactions using respective 3H-substrates and nonradioactive UDP-GlcNAc showed the presence of 2,4,6-tri-O-methylgalactose in the hydrolyzed products. This indicated the presence of a (1----3)-linked beta-D-GlcpNAc group at the nonreducing end in both cases. The linkage of the beta-D-GlcpNAc group to the subterminal D-Gal residue in the two products was confirmed by an almost 90% cleavage of the terminal [3H]GlcNAc group by purified clam and papaya beta-D-hexosaminidases.

Effect of nickel on testicular nucleic acid concentrations of rats on protein restriction.

The nucleic acids (DNA and RNA) and total protein concentration in testes were estimated in male Wistar strain rats treated intraperitorally with nickel sulfate (2.0 mg/100 g body weight) on alternate days for 10 dosages. In both normal (18% casein) and protein-restricted (5% casein) experimental animals, the nucleic acids and total protein concentration were found to decrease significantly compared to the corresponding controls. Sperm count and sperm motility were also reduced in both experimental groups of animals. The results indicate that nickel influences the expression of genetic information by reducing testicular nucleic acids and protein concentration in both dietary experimental groups.

Alteration of testicular biochemistry during protein restriction in nickel treated rats.

Nickel sulfate (2.0 mg/100 g.b.wt) dissolved in double-distilled water was administered (i.p.) on alternate days for ten doses to normal protein-fed and protein-restricted Wister strain albino rats (b.wt. 160 +/- 5 g). Two groups were used: one with normal protein diet, whereas the other with protein-restricted diet served as control. Twenty-four hours after the last treatment, the animals were sacrificed by decapitation. Tissues such as the testes, seminal vesicles, epididymis (Cauda and Caput) and prostate were dissected out, wiped clean, and stored at -20 degrees C until analysis. Lactate dehydrogenase (LDH) activities, glutamate oxaloacetate transaminase (GOT) activities, glycogen content, cholesterol content, and total protein content of the testes were estimated. Nickel sulfate administration significantly decreased the body weight of both normal protein-fed and protein-restricted groups of animals; the organ weights were also decreased. Significant decrease of LDH activity was observed, but GOT activity was not altered significantly. Testicular glycogen and cholesterol increased significantly in both experimental groups, but total protein content decreased. Nickel sulfate seems to have an adverse effect on the male reproductive system in both groups of animals fed with normal protein (18% casein) diet and protein restricted (5% casein) diet.

The influence of ascorbic acid on nickel-induced hepatic lipid peroxidation in rats.

We studied the effect of oral ascorbic acid treatment on nickel sulfate-induced lipid peroxidation in the liver of Wistar strain male albino rats. Lipid peroxide and glutathione levels and the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were estimated in liver. Nickel sulfate administration significantly increased the level of lipid peroxides and decreased glutathione, SOD, CAT, and GSH-Px activities in liver. The simultaneous administration of ascorbic acid with nickel sulfate resulted in a remarkable improvement of lipid peroxide, glutathione, SOD, CAT, and GSH-Px status in liver in comparison with rats treated with nickel alone. Nickel sulfate has an adverse effect on hepatic lipid peroxidation in animals, but simultaneous treatment with ascorbic acid offers a relative protection against nickel-induced hepatotoxicity.

Disability pattern amongst leprosy cases in an urban area (Calcutta).

In a retrospective study of 1,264 leprosy cases, registered during 1987-1992, 282 were found to have disabilities giving a disability rate (DR) 22.31% and 150 of them were also found to have deformities, giving a deformity rate 11.9%. Mean disability index (DI) was found to be 1.17. Disability rate (DR) significantly increased with age and the highest rate was 52.75% in lepromatous (L) cases, followed by 27.51% in borderline (N?L) and only 4.53% in nonlepromatous (N) cases. L cases had the highest deformity rate (22.25%) and N cases had the lowest DR (2.23%). DI was highest (1.46) in L, and lowest (0.52) in N cases. Males had significantly higher DR (27.2%) compared to females (13.0%). Deformity in hands (42.55%) was more common than in feet (22.70%). Increasing trend of DI was noticed with increasing duration of disease in L and N?L types. The number of nerves involved was high (4.72) in L cases compared to other types. DI was highest (1.25) in patients engaged in occupations involving hard work.

Study of the characteristics and causes of relapse amongst leprosy cases in an urban area (Calcutta).

In this retrospective study of the 3737 cases of leprosy released from treatment and followed-up during 1975 to 1990, 63 had relapsed giving an overall relapse rate of 1.69%. The relapse rate was significantly higher in the immunologically unstable N?L (Borderline) cases (2.9%). It was also higher in those who had dapsone monotherapy (1.92%) compared to those who had multidrug therapy (1.01%). The relapse rate was higher in the 10 to 29 years age group and among those who became pregnant suggesting puberty and pregnancy could be risk factors. Males had a significantly higher relapse rate (2.1%) than females (1.1%). 45.2% of relapses in N (Non-lepromatous) cases occurred within 24 months and 71.4% within 36 months of stopping treatment. In those having monotherapy, 57.1% of relapses occurred within 24 months and 76.8% within 36 months. Regularity in treatment did not seem to have much influence on relapse rates.

Influence of ascorbic acid on acid and alkaline phosphatase activities in some metabolically active tissues of aspirin treated rats.

ACP and ALP activities in plasma were increased in aspirin treated groups for a period of seven days. Ascorbic acid supplemented groups showed no significant change in plasma ACP activity, but a significant change in ALP activity was found. ACP and ALP activities in liver and kidney were decreased significantly in aspirin treated animals. ACP activities in liver and kidney in ascorbic acid supplemented groups showed no significant changes. No significant alteration of ALP activity in liver was found in ascorbic acid supplemented group but a significant changes was observed in kidney. Supplementation of ascorbic acid in high doses to rats fed aspirin can restore enzyme activities almost to the normal level.

Physical fitness: a longitudinal study among Muslim children of Bijapur (Karnataka).

Aerobic capacity or maximum oxygen uptake capacity (VO2 max) has been widely considered to be reliable and valid measure of cardio respiratory fitness. Persons possessing higher values and have the capacity to yield larger amounts of energy, are capable of performing better in athletic and other field activities. Seventy school going children from the Muslim community of Bijapur (Karnataka) aged 12-16 years (means +/- SEM = 14.33 +/- 0.94), volunteered for this study. Their height (cm) and weight (kg) were measured as physical anthropometry and Body mass index (BMI) was calculated (kg/m2). VO2max (ml.kg-1.min-1) was determined by applying the step test study of Margaria et al. The Physical fitness index (PFI) of the subjects were assessed by Harvard Step Test. The physiological endurance measured as VO2max (ml.kg-1.min-1) was found to be 34.31 +/- 2.44 S.E.M, which is lower in comparison to their Caucasian counterparts but nearly similar when compared with their Indian counterparts. The present study reveals that VO2max significantly correlates with BMI and PFI score. The present study also reveals that 27.2%, 20.07%, 15.77%, 14.37% and 22.87% of the subjects are in excellent, very good, good, average and poor classifications of fitness level respectively.

ROLE OF ASCORBIC ACID ON TRANSAMINASE ACTIVITIES IN SOME METABOLICALLY ACTIVE TISSUES OF ASPIRIN TREATED RATS.

Aspartate amino transferase (GOT) and alanine amino transferase (GPT) activities were studied in plasma, liver and kidney of aspirin treated and ascorbic acid supplemented groups for a period of seven days. GOT and GPT activities were increased in plasma but decreased significantly in liver and kidney in aspirin treated animals. Ascorbic acid supplemented groups showed no significant change of GOT and GPT in plasma and liver. In case of kidney, GOT activity remained unchange but GPT activity showed significant change in ascorbic acid supplemented group. The results clearly indicate that aspirin is a potent hepatotoxic and nephrotoxic drug but supplementation of ascorbic acid in High doses to rats fed aspirin can restore enzyme activities to the normal level.

Effect of a fatty acid moiety of phospholipid and ceramide on purified GalT-3 (UDP-Gal:GM2 beta 1-3 galactosyltransferase) activity from embryonic chicken brain.

Galactosyltransferase, GalT-3 (UDP-Gal:GM2 beta 1-3 galactosyltransferase) has been characterized and solubilized from 19-day-old embryonic chicken brain, and purified to over 2000-fold using mixed-modal chromatography on a omega-aminohexyl Sepharose column and affinity chromatography on a UDP-hexanolamine Sepharose column. The activity of purified GalT-3 was modulated by phospholipids in vitro with stimulation observed specifically with dipalmitoyl phosphatidylethanolamine (PE). All natural phospholipids tested (PE, PC and PI) inhibited GalT-3 activity. Enzyme activity was affected by the structure of the phospholipid vesicle. It was stabilized by the hexagonal (dipalmitoyl PE) structure and inhibited by the bilayer (dielaidoyl PE) structure. The long-chain fatty acid moiety of the glycosphingolipid substrate, GM2, was found to be necessary for optimum enzyme activity. In the absence of fatty acid, the modified substrates, lyso-GM2 and acetyl-GM2, had a 10-fold increased Km and a 4-8 fold decreased Vmax compared to the normal substrate. We postulate that GalT-3 belongs to a group of glycosyltransferases having recognition for both the carbohydrate as well as the hydrophobic domains (HY-CARS) of their substrates and that the fatty acid moiety of either the substrate (GM2) or a heterotropic effector (phospholipid) plays an important role in regulating the activity of this enzyme.

Efficacy of methotrexate in rheumatoid arthritis.

Rheumatoid arthritis is a common inflammatory articular disorder in Bangladesh. Methotrexate has proved to be an effective and relatively safe disease modifying drug for this disease. A quasiexperimental trial of the efficacy of methotrexate in rheumatoid arthritis was carried out in the Rheumatology Clinic, Institute of Postgraduate Medicine & Research, Dhaka during the period between July 1992 and September 1993. Thirty eight patients fulfilling the revised ARA criteria were given methotrexate in a total weekly dose of 7.5 to 15 mg. They were followed up at weekly intervals for one month and then monthly for a total duration of six months. Twenty three subjects eventually completed the trial. The trial showed significant differences in the disease activity indices at the end of six months. The decline of activity was noted at the end of one month. As a whole the response was complete in 4(17%), marked in 14(61%), moderate in 4(17%) and nil in 1(4%). Adverse effects occurred in 27 subjects. They were mild and transient in 22. Methotrexate appeared to be an acceptable DMARD for our rheumatoid arthritis population.