Effect of Leflunomide-Metal Complexes on ROS, TNF, and Brain Indolamines in Comparison with Anti-Depressants as Adjunct Therapy in Rheumatoid Arthritic Model.

Leflunomide is an isoxazole immunomodulating drug used to treat rheumatoid arthritis (RA). It is adopted as a metal-containing molecule to proceed with saturated salts of essential and detected metals; it amends the pharmacokinetic and pharmacodynamics activity of leflunomide to provide [M(Lef)4]X2-type complexes. Earlier it has been reported that after forming complexes with metals, leflunomide anti-arthritic activity was significantly altered in an acute arthritic model. In the present study, we evaluated the possible modification in anti-arthritic activities of leflunomide-metal complexes (Mg+2, Ca+2, Fe+2, Zn+2) with and without an anti-depressant drug, i.e., fluoxetine (10 mg/kg) in a chronic AIA model. Rats (n = 5) were administered with 0.1 mL of CFA into the right hind paw while treated groups received leflunomide and its metal complexes orally (3.2 mg/kg) for 24 days. On the final day of experiment, rats were sacrificed; a specific rat immunoassay ELISA kit was used to assess TNF-α in serum samples and read at 450 nm; a tissue sample of a paw was homogenized in a phosphate buffer using DCFH-DA dye for binding to assess ROS. A rat's brain sample was homogenized and evaluated for tryptophan, serotonin (5-HT), and HIAA by RP-HPLC with EC detector. The overall TNF production was altered in treated rats. In addition, a decreased ROS was observed in all categories, except lef+Mg+2 group. Moreover, depletion in the brain indolamine levels were found in treated groups; an upraised level of these indolamines was observed when fluoxetine was added. It is concluded that metals affect leflunomide activity on complexation and simultaneous administration of fluoxetine cope up with the depression in arthritic-induced rats.

Pharmacological effects of oral vitamin D intake on pulmonary function Test and Interleukin-17 in adult asthmatic patients in Karachi.

Vitamin D is an anti-inflammatory and immuno-modulatory secosteroid. Previous studies showed strong link between childhood and adult onset asthma with vitamin D. Interleukin 17 is an inflammatory cytokine and plays a major role in the worsening of asthma. The aim of this study is to compare the effects of serum vitamin D on serum IL 17 and pulmonary function test (FVC, FEV1, FEV1/FVC) before and after oral vitamin D supplementation. Fifty severe asthmatic patients were selected from out patient department of Chest Medicine Ward, Jinnah post graduate medical center, Karachi. Spirometry was performed by vitalograph compact. Baseline values were as follows: serum vitamin D=13.19±2.37ng/ml, IL-17=20.70±2.13ng/ml, FVC=2.31±0.40L, FEV1=1.40±0.28L,FEV1/FVC=60.15±4.61%. Subjects were given 1000 IU of oral vitamin D capsule per day for six weeks. After this trial all values were found as serum vitamin D=19.03±1.26ng/ml (p<0.001), IL 17=15.40ng/ml (p<0.001), FVC=2.90±0.60 L (p<0.001), FEV1=2.01±0.10L (p<0.001), FEV1/FVC=63.79% ±1.45 (p<0.001). It may be concluded that improvement in serum vitamin D levels improves the status of lung functions, decreases the airway inflammation and hence may decrease the asthma severity.

Inhibition of diet-restriction-induced behavioral deficits by tryptophan administration in rats.

Diet has a great impact on brain health and function. It plays an important role to improve and control a number of psychiatric disorders such as depression, anxiety, hyperactivity and behavioral impulsivity. Anorexia Nervosa (AN) is one of the psychiatric disorder which is associated with diet. In AN, patients show extreme dieting, weight loss, hyperactivity, depression/anxiety, self-control and behavioral impulsivity. Previous studies showed that during diet restriction, tryptophan decreases serotonin (5-hydroxytryptamine; 5-HT) metabolism in the brain due to its less availability and contributes psychiatric problems associated with AN. The present study is designed to investigate the effects of tryptophan administration on 5-HT metabolism in diet-restricted rats. Tryptophan at a dose of 50 or 100mg/kg was given orally to respective freely fed (FF) or diet restricted (DR) animals daily for five weeks. Behavioral activities were also monitored weekly. The results show significant effect (p<0.05) on behavior in activity box, open field and in light/dark transition test by tryptophan administration in diet-restricted rats. This may be associated with the increased in serum tryptophan and brain 5-HT metabolism. Therefore, it is concluded that diet-restriction-induced behavioral changes might be reverted back with the administration of tryptophan and may be helpful to improve psychological problems in AN.

Determination of tryptophan in raw materials, rat brain and human plasma by RP-HPLC technique.

This paper describes tryptophan (TRP) estimation in raw human plasma and rat brain by reversed-phase high-performance liquid chromatography (RP-HPLC). Estimation was carried out on a Purospher STAR C18 column using water-acetonitrile (90:10 v/v, at pH 2.7) mixture at a rate of 1.5 mL/min as mobile phase. Eluents were monitored at 273 nm by an ultraviolet detector. The method was linear (R(2) > 0.999), precise (intra-day and inter-day precision <2%) in the range of 0.25-20 µg/mL. The detection and quantification limits were 0.0144 µg/mL and 0.0437 µg/mL, respectively. In human plasma, Day 1 and Day 2 precision were 0.054-2.29% and 1.66-3.7%; whereas precisions in rat brain were 1.23-2.3% and 0.677-4.2%, respectively. The method was applied to study TRP level in human smokers and in arthritic rat brain. An efficient RP-HPLC method was developed for TRP determination that worked for clinical and research purposes.