Depletion of GSH in human blood plasma and cytosolic fraction during cadmium toxicity is temperature and pH dependent

Toxicities of heavy metals is a burning issue and a topic of interest among the toxicologists throughout the world. Metals are always in use of man since long but in recent years the use of cadmium has increased in the form of various cadmium compounds such as cadmium compounds as stabilizers in plastic pipe industries and in the preparations of different alloys etc. Cadmium is even used in phosphate fertilizers and thus comes directly or indirectly in contact with human eatables like crops, vegetables and fruits. Once it is absorbed it affects almost all the organs and systems of human body especially blood components and kidneys. Always the chemical reactions of different chemicals are dependent on some influential factors, among these factors the effect of pH and temperature of the media in which these chemicals interact with each other are very much important. Keeping in view this fact we have evaluated the effect of cadmium nitrate tetra hydrate on GSH of human plasma and cytosolic fraction. Estimation of thiol was done by Ellman's modified method and was found that the interaction of cadmium nitrate tetra hydrate and GSH of these blood components was more at a pH and temperature, which were near to physiological pH and temperature of human body. This fact was proved as the estimated thiol concentration left after the interaction of cadmium nitrate tetra hydrate and thiol of these blood components was minimum at pH and temperature near to human blood pH and temperature. We concluded that the possible reason for depletion of GSH of these blood components was conversion of GSH into Cd[SG][2]and/or GSSG formation

Unraveling the molecular mechanism governing the tissue specific expression of IFN-lamda-R1

The functional receptor for type III interferons [IFNs] is a heterodimer of IFNLR1 and IL10R2. IFNLR1 is expressed in a highly tissue specific manner, with epithelial and liver tissue as the prime expressing tissues in humans. However, knowledge about the molecular pathways responsible for regulating the expression of IFNLR1 is yet unknown. In this study, various bioinformatics tools were used to predict the scores of signal peptides of IFN-lamda-R1 and IFN-lamda-R1, which was considered as an important difference in the expression of both receptors or participation in regulating the IFNLR1 gene. In silico study revealed that the signal peptide of IFN-lamda-R1 had more potential than the signal peptide of IFN-lamda-R1 but changing the signal peptide of wild type IFN-lamda-R1 with the signal peptide of IFN-lamda-R1 in wet lab had barely shown any differences. Selective expression of IFN-lamda-R1 was considered to be a plus point towards the targeted anti-viral activity of IFN-lamda-s but artificial control on its expression will surely make IFN-lamda-s a better drug with enhanced activity. The results of this study may help us in contributing some understanding towards the mechanisms involved in the selective expression of IFNLR1 and exceptionalities involved

Effect of organo and inorganic lithium salt on human blood plasma glutathione- a comparative study

Investigation of toxicological effect of various metals is the field of interest for toxicological scientists since four to five decades and especially the toxicological effect of those drugs containing metals and there use is common because there is no other choice except to use these metal containing drugs. Inorganic as well as organic salts of lithium are commonly used in prophylaxis and treatments of many psychiatric disorders. The aim of the present study was to see the difference between the effect of organic and inorganic salt of lithium commonly used in psychiatric disorders on the GSH of human blood plasma. It is the scientific fact that ionic dissociation of organic and inorganic salts of any metal is always quite different hence to prove this fact, the effect of lithium citrate [organic salt of lithium] and lithium carbonate [inorganic salt of lithium] was investigated on human blood plasma GSH to find the difference between the effect of two. Ellman's method was used for the quantification of glutathione contents in plasma. It was found that lithium citrate decrease plasma GSH contents less than lithium carbonate indicating that organic salts of lithium are safe than inorganic salts of lithium when are used in psychiatric disorders. Further to analyze the effect of organic and inorganic salt of lithium on blood plasma GSH with the increase in incubation time was also evaluated and was found that both concentration and time dependent effect of organic salt of lithium shows that this salt has decreased plasma GSH contents of human blood less than inorganic salt of lithium either by promoting oxidation of GSH into GSSG or by lithium glutathione complex formation. These results suggest the physicians that the use of organic lithium salts is much safer than inorganic salts of lithium in terms of depletion of blood plasma GSH contents

Study of the effect of inorganic and organic complexes of arsenic metal on the status of GSH in T. cells and B. cells of blood

Arsenic is a major threat to large part of the population due to its carcinogenic nature. The toxicity of Arsenic varies with its chemical form and oxidation states. Glutathione [GSH], a major intra-cellular tripeptide plays a major role in arsenic detoxification. The present study was designed to provide insight into the extent of changes in GSH level by inorganic arsenic in the form of Arsenic trioxide [ATO] and organic arsenic in the form of nitro benzene arsenic acid [NBA]. Lymphocytes [T.cells and B.cells] were investigated for determination of change in GSH metabolic status caused by arsenic. The depletion of GSH level positively correlated with increasing arsenic concentration and time of incubation. The decline in GSH level was consistent with increasing pH and physiological temperature. Our findings show that changes in GSH status produced by Arsenic could be due to adduct [As-[SG][3]] formation. This change in GSH metabolic status provides information regarding mechanism of toxicity of inorganic and organic arsenicals. These findings are important for the rational design of antidote for the prevention of arsenic induced toxicity

Cadmium-glutathione complex formation in human t-cell and b-cell lymphocytes after their incubation with organo-cadmium diacetate

Cadmium intake is associated with oxidative stress that causes depletion of intracellular as well as extra cellular reduced glutathione. There is strong evidence indicating that reactive oxygen species and reactive nitrogen species generated in the presence of cadmium could be responsible for its toxic effects in many cells and tissues. Depletion of reduced glutathione in various cells, especially in T and B-lymphocytes, causes extreme damage to the antioxidant defense system of body. The aim of this research work was to investigate the metabolic changes that occur in T and B lymphocytes after their incubation with organ cadmium diacetate by using Ellman's spectrophotometric method of thiol quantification. The results of the present study indicate that cadmium depleted T and B lymphocytes GSH to a harmful extent. It is proposed that this depletion is due to the bivalent cadmium glutathione complex formation, oxidation of reduced glutathione [GSH] to its oxidized form, or both

simple and rapid approach to evaluate the in vitro in vivo role of release controlling agent ethyl cellulose ether derivative polymer

Diclofenac sodium [DCL-Na] conventional oral tablets exhibit serious side effects when given for a longer period leading to noncompliance. Controlled release matrix tablets of diclofenac sodium were formulated using simple blending [F-1], solvent evaporation [F-2] and co-precipitation techniques [F-3]. Ethocel[R] Standard 7 FP Premium Polymer [15%] was used as a release controlling agent. Drug release study was conducted in 7.4 pH phosphate buffer solutions as dissolution medium in vitro. Pharmacokinetic parameters were evaluated using albino rabbits. Solvent evaporation technique was found to be the best release controlling technique thereby prolonging the release rate up to 24 hours. Accelerated stability studies of the optimized test formulation [F-2] did not show any significant change [p<0.05] in the physicochemical characteristics and release rate when stored for six months. A simple and rapid method was developed for DCL-Na active moiety using HPLC-UV at 276nm. The optimized test tablets [F-2] significantly [p<0.05] exhibited peaks plasma concentration [C[max]=237.66 +/- 1.98] and extended the peak time [t[max]=4.63 +/- 0.24]. Good in-vitro in vivo correlation was found [R[2] =0.9883] against drug absorption and drug release. The study showed that once-daily controlled release matrix tablets of DCL-Na were successfully developed using Ethocel[R] Standard 7 FP Premium

Anti-dermatitis, anxiolytic and analgesic effects of Rhazya stricta from Balochistan

Current study was carried out on Rhazya stricta. Plant material was collected from Jhalmagsi Dist. Balochistan, Pakistan. Methanolic extract of Rhazya stricta was tested for anti-dermatitis, analgesic, anxiolytic effects, insecticidal activity and Brine shrimp Bioassay. Crude extract showed significant anti-dermatitis activity, as the results of intensity score showed mild Excoriation or erosion, moderate Edema or populations and absence of Erythema or hemorrhage, Scratching time was decreased to 1.45 and histological observations of mice treated with crude extract showed mild changes and few inflammatory cells in several microscopic fields. The results of analgesic activity were significant and the percentage inhibition of writhes were 73.54% and 69.38% at 300mg/kg and 500mg/kg respectively. The overall response of crude extract in anxiolytic activities were depressive and crude extract showed sedative effects. In Brine shrimp [Artemsia salina] lethality bioassay crude extract showed dose depended significant activity, and showed positive lethality with LD50 3.3004 micro g/ml. Insecticidal activity was positive against Callosbruchus analis, the percent mortality was 40%.

Change in metabolic status of glutathione by palladium nitrate in blood components

This piece of research work present the toxicological impact of varied concentrations of Palladium Nitrate [Pd [NO[3][2] by changing the chemical status of glutathione and the way how glutathione plays its role in detoxification and conjugation processes of [Pd [NO[3][2] in whole blood components [plasma and Cytosolic fraction]. The impact of different concentration of [Pd [N03]2] on reduced glutathione level in whole blood component [Plasma and Cytosolic fraction] were measured spectrophotometrically following Standard Ellman's method. Compared with control sample, significant decrease in the GSH content in whole blood components [plasma and Cytosolic fraction] was obtained with various concentrations [100microM-1000microM] of Palladium Nitrate. Depleted GSH level was more pronounced with time incubation period [0-90] minutes. These finding shows that changes in the GSH status produced by palladium nitrate could either be due to palladium nitrate and glutathione [Pd-SG] complex formation or by conversion of reduce glutathione [2GSH + Pd[+2] -> GSSG]. This change in the GSH metabolic status provides information regarding the mechanism of palladium, in blood components

Over expression of a synthetic gene encoding interferon lambda using relative synonymous codon usage bias in Escherichia coli

Interferon Lambda [IFN- Lamda] is a type III interferon which belongs to a novel family of cytokines and possesses antiviral and antitumor properties. It is unique in its own class of cytokines; because of the specificity towards its heterodimer receptors and its structural similarities with cytokines of other classes. This renders IFN- Lamda a better choice for the treatment against many diseases including viral hepatitis and human coronavirus [HCoV-EMC]. The present study describes a computational approach known as relative synonymous codon usage [RSCU]; used to enhance the expression of IFN- Lamda protein in a eukaryotic expression system. Manually designed and commercially synthesized IFN- Lamda gene was cloned into pET-22b expression plasmid under the control of inducible T7-lac promoter. Maximum levels of IFN- Lamda expression was observed with 0.4 mM IPTG in transformed E. coli incubated for 4 hours in LB medium. Higher concentrations of IPTG had no or negative effect on the expression of IFN- Lamda. This synthetically over expressed IFN- Lamda can be tested as a targeted treatment option for viral hepatitis after purification

Synthesis, conjugation and evaluation of some novel polymers and their micro particles for sustained release drug formulations

To prepare and evaluate three novels functionalized polymers [PGA, PGA-co-caprolactone and PGA-copentadecalactone] for the development of nanoparticles which were further used in the development of a novel polymeric prodrug using Ibuprofen as a model drug. The Ibuprofen-polymer prodrug was developed by coupling the drug to one of the three prepared polyester polymers via ester linkage. A hydrolytic enzyme was used to prepare two polymer monomers, glycerol and polyvinyl adipate, which are non toxic, ester linked biological monomers. The polymers and their prodrug were characterized using NMR, GPC, UV-spectrophotometer and DSC. In vitro drug release study of Ibuprofen-polymer conjugate was performed in phosphate buffer PH 7.4 using a roller [Stuart STR 1] placed in an incubator [Stuart SI 60] and the temperature was kept constant at 37 +/- 1°C. Among the three polymers, glycerol-adipateco-pentadecalactone was observed to give a burst release following slow release in the medium. These characteristics suggest that these polymers can be successfully used in sustained release drug formulations

Preparation and evaluation of [99m]Tc-DMSA lyophilized kit for renal imaging

Dimercaptosuccinic acid [DMSA] has been evaluated and used with technetium 99m [[99m]Tc] in imaging of kidneys. DMSA lyophilized kits were prepared and radiolabelled with [99m]Tc. Paper and thin-layer chromatography have been employed using various eluent systems for the radiochemical analysis, percentage labeling and binding capacity of [99m]Tc-DMSA. Female albino rabbits were used for this study. Biological data obtained after intravenous injection of radiolabelled DMSA to female albino rabbits revealed 32.42% uptake and long retention time in the kidneys. On the basis of animal biodistribution data, it is suggested that DMSA when labeled with [99m]Tc is useful complex for renal imaging and can be successfully applied as a diagnostic tool in nuclear medicine. Clinical biodistribution and radiation dosimetry studies are planned in future

Formulation and in vitro evaluation of clotrimazole gel containing almond oil and tween 80 as penetration enhancer for topical application

Achieving a desirable percutaneous absorption of drug molecule is a major concern in formulating dermal and transdermal products. The use of penetration enhancers could provide a successful mean for this purpose. The aim of this study was to develop Clotrimazole gel and to evaluate the effect of almond oil and tween 80 [in different concentrations], on the permeation of drug through rabbit skin in vitro. In order to investigate the effect of penetration enhancers used in this study on the permeation of Clotrimazole through sections of excised rabbit skin, Franz diffusion cell was employed. Sample solution was withdrawn at specific time interval up to 24 h. Significant difference in permeation among the eight formulations was seen in the study. The permeation profile of various formulations also showed that the added enhancers in individual batches affected the permeation of the drug. Drug permeation increased with increased concentration of Tween 80 and decreased concentration of almond oil. Furthermore, almond oil combined with tween 80 showed synergistic effect. The clotrimazole gels were successfully formulated and could be beneficial for topical use

Development of novel diclofenac potassium controlled release tablets by wet granulation technique and the effect of co-excipients on in vitro drug release rates

The aim of the present study was the formulation and evaluation of controlled release polymeric tablets of Diclofenac Potassium by wet granulation method for the release rate, release pattern and the mechanism involved in drug release. Formulations having three grades of polymer Ethocel [7P; 7FP, 10P, 10FP, 100P, 100FP] in several drugs to polymer ratios [10:3 and 10:1] were compressed into tablets using wet granulation method. Co-excipients were added to some selected formulations to investigate their enhancement effect on in vitro drug release patterns. In vitro drug release studies were performed using USP Method-1 [Rotating Basket method] and Phosphate buffer [pH 7.4] was used as a dissolution medium. The similarities and dissimilarities of release profiles of test formulations with reference standard were checked using f2 similarity factor and f1 dissimilarity factor. Mathematical/Kinetic models were employed to determine the release mechanism and drug release kinetics

Formulation development and investigation of ibuprofen controlled release tablets with hydrophilic polymers and the effect of co-excipients on drug release patterns

The aim and objective of the present study was to formulate and evaluate controlled release polymeric tablets of Ibuprofen with determinations of formulation factors using various grades and types of polymer Ethocel i.e. Ethocel Standard 10P; 10FP, 100P and 100FP for their release rates and release patterns in suitable media and also the mechanism involved in the release of drug from the matrices. The effect of several co-excipients was also studied on the drug release rates and patterns of Ibuprofen from the polymeric matrices. Ibuprofen-Ethocel CR tablets were prepared at three different D:P ratios i.e. 10:1, 10:2 and 10:3. The effects of co-excipients were studied only in formulations having D:P ratio of 10:3. In vitro drug release studies of Ibuprofen-Ethocel controlled release matrix tablets were carried out in phosphate buffer pH 6.8 using Pharma Test Dissolution Apparatus adopting Rotating Basket Method according to USP. Different kinetic models were applied to the release data of test formulations in order to investigate the release mechanism of drug from the controlled release matrix tablets. The release patterns of Ibuprofen-Ethocel CR matrices were compared with reference conventional Ibuprofen tablets and Ibuprofen SR tablets. F[2] similarity factor was applied to the test formulations and reference standard to compare their similarities. The drug formulations studied exhibited satisfactory release results

Study of the effects of BaCl[2] on the level of glutathione in plasma and cytosolic fraction in whole blood

Barium has important role in the field of medical sciences, but it has been found in various studies that barium can cause numerous toxic effects. Studies have proven the strong affinity of the metalloelements for the sulfhydryl group [SH], present in reduced glutathione [GSH] and other biological molecules. In this context, the study about the possible interaction of BaCl[2] with glutathione in whole blood components was of interest, as an indication about the extent of barium toxicity and the role of glutathione in the conjugation and detoxification of the metalloelement barium. The concentration dependent and time dependent effect of BaCl[2] on the level of GSH in plasma and Cytosolic Fraction in whole blood was investigated, following Ellman's method. It was found that BaCl[2] causes a decrease in the GSH level, which is more pronounced with increasing concentration of BaCl[2] and with time incubation as well. The observed effect GSH concentration may be presumably due to production of oxidized glutathione [GSSG] or then due to Barium-Glutathione [GS-Ba-SG] conjugate formation

Effect of olive oil on transdermal penetration of flurbiprofen from topical gel as enhancer

The present study was conducted to formulate and evaluate flurbiprofen transdermal gel. A standard calibration curve was constructed to obtain a regression line equation to be used for finding out the concentration of drug in samples. Olive oil was used as penetration enhancer and was added in different concentrations to some selected formulations to see its enhancement effect on in vitro drug release profiles. The prepared gels were evaluated for several physico-chemical parameters to justify their suitability for topical use. The in vitro drug release studies were carried out by using Franz cell diffusion apparatus across both synthetic membrane and excised albino rabbit skin. In order to investigate the drug release mechanism a kinetic approach was made by employing Korsmeyer kinetic model to the in vitro drug release profiles of flurbiprofen. The flurbiprofen topical gels were successfully prepared and could be beneficial for topical use

Role of glutathione in protection against mercury induced poisoning

Mercury is harmless in an insoluble form, such as mercuric sulfide, but it is poisonous in soluble forms such as mercuric chloride or methylmercury. Mercury is a neurotoxin. Outbreaks of mercuric chloride poisonings have made it clear that adults, children, and developing fetuses are at risk from ingestion exposure to mercury. It is very important and interesting to study the reaction of mercuric chloride and Glutathione as biomarker of Glutathione role in detoxification and conjugation in components [Plasma and Cytosolic Fraction]. The effect of mercuric chloride's different concentrations was examined on GSH present in plasma and cytosolic fraction. Decrease in GSH level was dependant on mercuric chloride concentration. The decrease in GSH level of blood components was more prominent with the time of incubation of mercuric chloride. Decrease in the concentration of reduced state Glutathione may be due the interaction of reduced state Glutathione [GSH] and mercuric chloride to form oxidized Glutathione [GSSG] or mercuric-glutathione complex. This change in GSH metabolic status provides information regarding the role of GSH in detoxification of mercuric chloride. The effect of mercury metal on Glutathione in blood components has been discussed in this paper in vitro condition as a model for in Vivo condition

Evaluation of the interaction of vanadium with glutathione in human blood components

Metallo-elements including Vanadium [V] have strong affinity for sulfhydryl [-SH] groups in biological molecules including Glutathione [GSH] in tissues. Because of this fact it was of interest to further investigate the interaction of Ammonium Vanadate [NH[4]VO[3]] with Glutathione as a biomarker of toxicity and the role of Glutathione in the detoxification and conjugation processes in whole blood components including plasma and cytosolic fraction. Effects of different concentrations of Ammonium Vanadate [NH[4]VO[3]] on the level of reduced Glutathione in whole blood components [Plasma and Cytosolic fraction] were examined. GSH depletion in plasma and cytosolic fraction was Ammonium Vanadate's concentration-dependent. Depleted GSH level was more pronounced with more incubation time period. These findings show that changes in the GSH status produced by Ammonium Vanadate could be due to either by adduct formation of Vanadium and glutathione i.e. [V-SG] or by increased production of oxidized Glutathione [2GSH +V[+5] - GSSG]. This change in GSH metabolic status provides some information regarding the mechanism of toxicity by Ammonium Vanadate and the protective role of glutathione

Effect of aluminium metal on glutathione [GSH] level in plasma and cytosolic fraction of human blood

Aluminium is being used in the medicines in the form of antacids. The Aluminium metal can be leached from our utensils and can harm the body for its side effects, if become available to the systemic circulation. So it is important to check the effect of Aluminum on the Glutathione in vivo condition. Ellman method was used to determine the effect of Aluminum on GSH level in whole blood spectrophotometerically. 5,5-Dithiobis, 2-Nitrobenzoic Acid, Glutathione, Aluminium sulphate, phosphate buffer, HC1 [Hydrochloric acid] and other laboratory instruments were used to conduct the research work. Time dependent effect of Aluminum on Glutathione level in whole blood was also checked and decrease was observed. This study also shows the effect of Aluminum as helping agent for the Glutathione to enhance the antioxidant system of the body or a cause for depletion of reduced Glutathione

Formulation and in vitro evaluation of ofloxacin-ethocel controlled release matrix tablets prepared by wet granulation method: influence of co-excipients on drug release rates

Being controlled release dosage forms, tablets allow an improved absorption and release profiles of Ofloxacin. The fact that drugs with fine particles size can be compressed well after wetting, so in our research studies Ofloxacin controlled release matrix tablets were prepared by wet granulation technique. In order to investigate the potential of Ethyl cellulose ether derivatives as a matrix material, Ofloxacin formulations with different types and grades of Ethocel were prepared at several drug-to-polymer ratios. The method adopted for in vitro drug release studies was USP Method-1 [rotating Basket Method] by Pharma test dissolution apparatus using phosphate buffer 7.4 pH as a dissolution medium. Various Kinetic models were employed to the formulations for the purpose of determination of release mechanism. A comparative study was performed between the tested Ofloxacin-Ethocel formulations and a standard reference obtained from the local market. FI dissimilarity factor and f2 similarity factor were applied to the formulations for the checking of dissimilarities and similarities between the tested formulations and reference standard