Neuroprotective effects of theobromine in transient global cerebral ischemia-reperfusion rat model.

Transient global cerebral ischemia (tGCI) is a cerebrovascular disorder characterized by a brief decline in blood flow, neurological deficits, and is often predictive of stroke. Theobromine (TBR) is consumed worldwide in chocolates, tea, and cocoa products. TBR is a natural stimulant and vasoactive alkaloid that may protect against ischemic injury. In this study, neuroprotective potential of theobromine (TBR) was evaluated in 2-vessel occlusion transient global cerebral ischemia-reperfusion (tGCI/R) rat model. Rats were treated with TBR (50, 100 mg/kg, p.o.) for 7 successive days, and subjected to bilateral common carotid artery occlusion (20 min) or sham surgery after last dose of TBR. Severe neurological deficits accompanied by brain infarction, blood-brain barrier abnormalities, and oedema were noted in rats subjected to tGCI/R, and these effects were prevented by TBR. TBR protected against lipid peroxidation and enhanced glutathione level in brain against tGCI/R. TBR pre-treatment for 7 days prevented tGCI/R induced cell death (lactate dehydrogenase, caspase-3), vascular injury (MMP-9), and inflammation (TNF-α, NFκB) in rat whole brain. TBR protected against glutamate excitotoxicity and GABAergic decline in the brain of rats against tGCI/R injury. Findings of this study showed that TBR can alleviate chances of stroke by preventing acute episodes of cerebral ischemia.

Arbutin protects brain against middle cerebral artery occlusion-reperfusion (MCAo/R) injury.

Focal ischemia causes irreversible brain damage if cerebral blood flow is not restored promptly. Acute phase excitotoxicity and pro-oxidant and inflammatory events in the sub-chronic phase elicit coagulative necrosis, vascular injury, cerebral oedema, and neurobehavioral deficits. Earlier, in pre-clinical studies arbutin protected behavioral functions and improved therapeutic outcomes in different models of brain and metabolic disorders. Arbutin is natural hydroquinone that might protect against ischemia-reperfusion (I/R) injury. In this study, cerebro-protective effects of arbutin were evaluated in the middle cerebral artery occlusion-reperfusion (MCAo/R) mouse model. Mice were administered arbutin (50, 100 mg/kg, i.p.) for 21 days, and subjected to MCAo/R or sham surgery on day 14. Results showed brain infarction, blood-brain barrier dysfunction, oedema, and neurological deficits 24 h post-MCAo/R injury that were prevented by arbutin. Behavioral evaluations over the sub-chronic phase revealed MCAo/R triggered spatial and working memory deficits. Arbutin protected the memory against MCAo/R injury and decreased hydroxy-2'-deoxyguanosine, protein carbonyls, inflammatory cytokines (tumor necrosis factor-α, myeloperoxidase, matrix metalloproteinase-9, inducible nitric oxide synthase), and enhanced glutathione levels in the ischemia ipsilateral hemisphere. Arbutin decreased brain acetylcholinesterase activity, glutamate, and enhanced GABA levels against MCAo/R. Arbutin can alleviate I/R pathogenesis and protects neurobehavioral functions in the MCAo/R mouse model.

Serum and Saliva LDH Levels in Patients with Habit, Oral Potentially Malignant Disorders (OPMDs), and Oral Squamous Cell Carcinoma (OSCC): A Spectrophotometry Study.

Background: Oral potentially malignant disorders (OPMDs) are chronic lesions or conditions characterized by a potential for malignant transformation. One of the hallmarks of cancer is aerobic glycolysis which confers immortality to cancer cells, correlates with the belligerent behavior of various malignancies, and shows an increase in the expression of lactate dehydrogenase (LDH). The present study was conducted to measure and compare serum and salivary LDH levels in patients with habit, OPMDs and oral squamous cell carcinoma (OSCC) patients, and to evaluate if LDH levels can be used as a biomarker in the progression into potentially malignant disorders (PMDs), the prognosis of OSCC, and to assess if saliva is a better biomarker. Materials and Methods: Thirty patients of both genders were divided into three groups. Group I had patients with habits, group II had OPMDs, and group III had OSCC with 10 subjects from each group. Saliva and serum of patients were collected according to the standard protocol and measured for LDH using spectrophotometry of 340 nm. Results: Group I had patients with habits, group II had OPMDs, and group III had OSCC. Group I had 6 males and 4 females, group II had 8 males and 2 females, and group III had 7 males and 3 females. The mean serum LDH level in group I was 265.50, in group II was 194.10, and in group III was 224.22. The difference was non-significant (P > 0.05). The mean salivary LDH level in group I was 345.68, in group II was 532.72, and in group III was 1.105. The difference was significant (P < 0.05). Conclusion: Salivary LDH is a non-invasive, cost-effective technique, which can be used as an effective modality for the diagnosis and prognosis of oral cancer as well as for monitoring tobacco users and OPMDs.

Artesunate-loaded chitosan/lecithin nanoparticles: preparation, characterization, and in vivo studies.

Artesunate (AST), the most widely used artemisnin derivative, has poor aqueous solubility and suffers from low oral bioavailability (~40%). Under these conditions, nanoparticles with controlled and sustained released properties can be a suitable solution for improving its biopharmaceuticals properties. This work reports the preparation and characterization of auto-assembled chitosan/lecithin nanoparticles loaded with AST and AST complexed with ß-cyclodextrin (ß-CD) to boost its antimalarial activity. The nanoparticles prepared by direct injection of lecithin alcoholic solution into chitosan/water solution have shown the particle size distribution below 300 nm. Drug entrapment efficiency was found to be maximum (90%) for nanoparticles containing 100 mg of AST. Transmission electron microscopy images show spherical shape with contrasted corona (chitosan) surrounded by a lipidic core (lecithin + isopropyl myristate). Differential scanning calorimeter thermograms demonstrated the presence of drug in drug-loaded nanoparticles along with the disappearance of decomposition exotherm suggesting the increased physical stability of drug in prepared formulations. Negligible changes in the characteristic peaks of drug in Fourier-transform infrared spectra indicated the absence of any interaction among the various components entrapped in the nanoparticle formulation. In vitro drug release behavior was found to be influenced by pH value. Increased in vivo antimalarial activity in terms of less mean percent parasitemia was observed in infected Plasmodium berghei mice after the oral administration of all the prepared nanoparticle formulations.

Exploring the potential of lecithin/chitosan nanoparticles in enhancement of antihypertensive efficacy of hydrochlorothiazide.

The present study involves the preparation of lecithin/chitosan nanoparticles loaded with hydrochlorothiazide (HCT) (a poorly water soluble antihypertensive) and hydrochlorothiazide complexed with ß-cyclodextrin (HCT-ß-CD) with a view to improve its biopharmaceutical properties. Nanoparticles prepared using modified solvent evaporation method showed a particle size in the range of 126-139 and 152-181 nm (polydispersity index, PDI = 0.2) for HCT and HCT-ß-CD loaded nanoparticles respectively. TEM images revealed their spherical nature. The stable nature of the prepared formulations was evident from the high positive value of zeta potential (>20 mV). HCT and HCT-ß-CD loaded nanoparticles with 150 mg of drug have shown a maximum entrapment efficiency of 81.8 ± 1.7% and 91.1 ± 1.5% respectively. In vitro studies have shown an improved and a sustained release pattern. In vivo activity in DOCA induced hypertensive rats demonstrates 1.5-fold percentage decrease in systolic blood pressure and a prolonged duration of action.

Comparative analysis of genome-based CAZyme cassette in Antarctic Microbacterium sp. PAMC28756 with 31 other Microbacterium species.

BACKGROUND: The genus Microbacterium belongs to the family Microbacteriaceae and phylum Actinobacteria. A detailed study on the complete genome and systematic comparative analysis of carbohydrate-active enzyme (CAZyme) among the Microbacterium species would add knowledge on metabolic and environmental adaptation. Here we present the comparative genomic analysis of CAZyme using the complete genome of Antarctic Microbacterium sp. PAMC28756 with other complete genomes of 31 Microbacterium species available. OBJECTIVE: The genomic and CAZyme comparison of Microbacterium species and to rule out the specific features of CAZyme for the environmental and metabolic adaptation. METHODS: Bacterial source were collected from NCBI database, CAZyme annotation of Microbacterium species was analyzed using dbCAN2 Meta server. Cluster of orthologous groups (COGs) analysis was performed using the eggNOG4.5 database. Whereas, KEGG database was used to compare and obtained the functional genome annotation information in carbohydrate metabolism and glyoxylate cycle. RESULTS: Out of 32 complete genomes of Microbacterium species, strain No. 7 isolated from Activated Sludge showed the largest genomic size at 4.83 Mb. The genomic size of PAMC28756 isolated from Antarctic lichen species Stereocaulons was 3.54 Mb, the G + C content was 70.4% with 3,407 predicted genes, of which 3.36% were predicted CAZyme. In addition, while comparing the Glyoxylate cycle among 32 bacteria, except 10 strains, all other, including our strain have Glyoxylate pathway. PAMC28756 contained the genes that degrade cellulose, hemicellulose, amylase, pectinase, chitins and other exo-and endo glycosidases. Utilizing these polysaccharides can provides source of energy in an extreme environment. In addition, PAMC28756 assigned the (10.15%) genes in the carbohydrate transport and metabolism functional group closely related to the CAZyme for polysaccharides degradation. CONCLUSIONS: The genomic content and CAZymes distribution was varied in Microbacterium species. There was the presence of more than 10% genes in the carbohydrate transport and metabolism functional group closely related to the CAZyme for polysaccharides degradation. In addition, occurrence of glyoxylate cycle for alternative utilization of carbon sources suggest the adaptation of PAMC28756 in the harsh microenvironment.

Variability in serum electrolytes in different grades of depression.

The present study was planned to assess the variation in serum electrolytes in patients of major depression. A total of 100 patients of 35-45 yrs of age (68 males and 32 females) of depression were compared with the age matched healthy volunteers. Severity of depression was assessed by DSM IV criteria and were graded into mild, moderate and severe depression. In all the subjects serum electrolytes (Na+, K+, Mg++ and Ca++) were assessed quantitatively. All the depression patients were having higher level of Na+, K+, and Ca++ and lower level of Mg++. Multivariate analysis showed that different grades of depression influences statistically significantly levels of serum Mg++ and Ca++ (F(2,98) for Mg++ = 4.88, P value = 0.001 and F(2,98) for Ca++ = 5.97, P value = 0.004). No statistically significant difference was observed for Na+ and K+ although their values were higher (within normal limit) in comparison to that of control group. Multiple comparison revealed highly statistically significant difference between the levels of serum Ca and Mg individually between mild and moderate depression (S V(mild & mod) for Ca++; Mg++ = 7.1; 7.4) moderate & severe depression (SV(mod & severe) for Ca++; Mg++ = 6.82; 6.92) and mild & severe depression patients (SV(mild & severe) for Ca++; Mg++ = 7.46; 7.73) with critical value = 6.36. These results indicated the disbalance in the level of serum electrolytes especially of Ca and Mg in accordance with the severity of depression.

Treatment of Chronic Hepatitis due to Hepatitis C Virus (CH-C) in India: A Randomized Controlled Trial Comparing Daily Interferon-alfa-2b and Ribavirin with Daily Interferon-alfa-2b and Glycyrrhizin-A Multicenter Study.

BACKGROUND AND AIM: Pegylated-interferon-alfa (PEG-IFN-α) with ribavirin is an established treatment in chronic hepatitis due to hepatitis C virus (HCV) (CH-C). Such treatment is expensive and in resource-poor countries such as India, alternative less expensive therapy is needed. METHODS: Multicenter randomized controlled trial comparing two treatment regimens (interferon-alfa-2b [IFN-α-2b] 3 million unit/day [MU/day] and ribavirin 1000 mg/day [I+R] vs IFN-α-2b 3 MU/day and glycyrrhizin 250 mg [I+G]) in CH-C. Viral, host characteristics and therapeutic responses were assessed (ICMR-6 months trial for chronic hepatitis-CTRI/2008/091/000105). RESULTS: One hundred and thirty-one patients meeting the inclusion criteria were randomized to I + G (n=64) or I+R (n=67) during the period February 2002 to May 2005. About 85% (I+G=53, I+R=58) completed 6 months of treatment and 89% of them (I+G=46, I+R=53) completed 6 months of follow-up after completion of treatment. Hepatitis C virus genotype 3 was the major type detected (71% patients). The mean log10 viral load (copies/mL), histological activity index, and fibrosis stage for all patients were 5.1 ± 0.98, 5 ± 2, and 2± 1.5, respectively. Sustained viral response (SVR) was significantly higher in I + R group than in I + G group (65.7% vs 46.9%, OR=2.2, P = 0.03). Treatment with I + G was associated with significantly lower frequencies of leukopenia (2% vs 17%, P <0.01) and anemia (8% vs 40%, P <0.001) as compared to treatment with I + R. CONCLUSION: Genotype 3 HCV infection with low viral load is prevalent in India. Daily IFN with ribavirin showed significantly better responses. Leukopenia and anemia were significantly more in ribavirin group. Responses observed with IFN + ribavirin were similar to the reported response rates with PEG-IFN suggesting that this modality may be considered as a cheaper alternative of treatment for chronic hepatitis C.

Interaction of artesunate with ß-cyclodextrin: Characterization, thermodynamic parameters, molecular modeling, effect of PEG on complexation and antimalarial activity.

Inclusion of artesunate in the cavity of ß-cyclodextrin (ß-CD) as well as its methyl and hydroxypropyl derivatives was investigated experimentally and by molecular modeling studies. The effect of PEG on the inclusion was also studied. A 1:1 stoichiometry was indicated by phase-solubility studies both in the presence and absence of PEG and suggested by the mass spectrometry. The mode of inclusion was supported by 2D NMR and results were further verified by docking studies utilizing Fast Rigid Exhaustive Docking acronym. The thermodynamic parameters were determined for both binary and ternary systems using solution calorimetry and were found to be best for the methyl-ß-cyclodextrin (Me-ß-CD) system. However, the presence of PEG improves the complexation ability as evident from elevation in the numerical value of the stability constant (K). Solubility and dissolution profile of binary complex is enhanced in the presence of PEG, which is approximately at par with drug Me-ß-CD complexes. In vivo studies showed 100% survivability in artesunate-Me-ß-CD complexes.