Salivary Biomarkers for Periodontal Diseases-A Review.

Second most common oral disease next to dental caries is periodontal disease. It is considered to be inflammatory disorder that damages tissue through the complex interaction between periopathogens and the host defense systems. Researchers involved in periodontal disease diagnostics are currently investigating the possible use of oral fluids, such as saliva, for disease assessment. Secretions from the major salivary glands, which have a large number of proteins and peptides, are responsible for maintaining the integrity of the oral cavity. Also, because of its importance in oral biofilm formation and host defense, secreted saliva with its biomarkers may have a significant role in the establishment and progression of periodontal disease.

Evaluation of disease modifying activity of Coriandrum sativum in experimental models.

Background & objectives: Coriandrum sativum (CS), has been widely used in traditional systems of medicine for treatment of rheumatoid arthritis. However, the mechanism of action for its antiarthritic effects is not clearly known. Therefore, the present study was carried out to evaluate the antiarthritic activity of CS in rats in two experimental models. Methods: The antiarthritic activity of CS seed hydroalcoholic extract (CSHE) was evaluated in adult Wistar rats by using two experimental models, viz. formaldehyde and Complete Freund's adjuvant (CFA) induced arthritis. The expression of pro-inflammatory cytokines (predominantly contributed by macrophages) was also evaluated. TNF-α level was estimated in serum by ELISA method. TNF-R1, IL-1 β and IL-6 expression in the synovium was analysed by immunohistochemistry. Results: CSHE produced a dose dependent inhibition of joint swelling as compared to control animals in both, formaldehyde and CFA induced arthritis. Although there was a dose dependent increase in serum TNF-α levels in the CSHE treated groups as compared to control, the synovial expression of macrophage derived pro-inflammatory cytokines/cytokine receptor was found to be lower in the CSHE treated groups as compared to control. Interpretation & conclusions: Our results demonstrate that the antiarthritic activity of CSHE may be attributed to the modulation of pro-inflammatory cytokines in the synovium. In further studies CSHE could be explored to be developed as a disease modifying agent in the treatment of RA.

A pilot study on area under curve of mycophenolic acid as a guide for its optimal use in renal transplant recipients.

Background & objectives: The immunosuppressants administered to renal transplant subjects are usually monitored therapeutically to prevent graft rejection and drug toxicity. Mycophenolic acid (MPA) is an immunosuppressant. The present prospective study was undertaken to establish the utility of plasma level monitoring of MPA and to correlate it with clinical outcomes in renal transplant receipients. Methods: MPA plasma level at 2, 4 and 9 h and the area under concentration-time curve (AUC) were estimated using high performance liquid chromatography in 24 renal transplant recipients receiving immunosuppressant MPA plus tacrolimus and steroid. Results: There was wide inter-individual variation in MPA plasma level and the AUC. The incidences of gastrointestinal adverse drug events (diarrhoea and acidity) were significantly more in the high MPA AUC patients. Though biopsy proven acute rejection was not found, of the six subjects with lower MPA AUC (<30 mg.h/l), three were clinically diagnosed to develop tacrolimus nephrotoxicity. The Gastrointestinal Symptom Rating Scale (GSRS) and Gastrointestinal Quality of Life Index (GIQLI) scores represented better health related quality of life in lower MPA AUC than in the higher MPA AUC (>60 mg.h/l). Interpretation & conclusions: The present findings suggest the MPA AUC of 30 - 60 mg.h/l in the maintenance stage of renal transplant patients to have optimum clinical benefit and relegated adverse events profile indicating the usefulness of AUC of MPA with limited sampling strategy in optimizing its use.

Antiarthritic activity of Majoon Suranjan (a polyherbal Unani formulation) in rat.

Background & objectives: Majoon Suranjan (MS) is a polyherbal formulation used in Unani system of medicine for the treatment of rheumatoid arthritis (RA). The present study evaluates the antiarthritic efficacy of this formulation in three different experimental models. Methods: The anti-inflammatory activity of MS (in doses of 450, 900 and 1800 mg/kg body wt) was evaluated using the turpentine oil induced paw oedema model and the antiarthritic efficacy was evaluated using the formaldehyde and complete Freund's adjuvant (CFA) induced arthritis models. Aspirin (100 mg/kg body wt) was used as the standard drug in all the models. In order to assess the safety of the test drug, oral acute and 28 day toxicity studies were also carried out. Results: MS produced a dose dependent protective effect in all the experimental models. Its antiarthritic efficacy was comparable to aspirin in formaldehyde induced arthritis and was superior to aspirin in turpentine oil induced paw oedema and CFA induced arthritis. MS also inhibited the delayed increase in joint diameter as seen in control and aspirin treated animals in CFA induced arthritis. Oral LD50 of MS was found to be >5000 mg/kg in rats. Chronic administration did not produce any significant physiological changes in the tested animals. Interpretation & conclusions: Results of the present study suggest that the antiarthritic activity of MS was due to the interplay between its anti-inflammatory and disease modifying activities, thus supporting its use in traditional medicine for the treatment of RA.

Gaso-transmitter hydrogen sulphide: Potential new target in pharmacotherapy.

Research in the last two decades has transformed the way hydrogen sulphide (H2S) is perceived from a noxious gas to a gaso-transmitter with a vast potential in pharmacotherapy. H2S is synthesized in various body-systems using the enzymes cystathionine beta-synthase and cystathionine gamma-lyase; either of these being the predominat enzyme in a particular system. H2S may be one of the physiological modulators of blood pressure in humans. The gas relaxes the vascular smooth muscle cells by opening up KATP channels. Moreover, it suppresses the proliferation of vascular smooth muscle cells. H2S may also be contributing in the protection afforded by ischaemia-preconditioning. Testosterone is thought to be responsible for the higher central nervous system level of H2S in males. In the central nervous system, H2S is implicated in Alzheimer’s disease, epilepsy, stroke and Down’s syndrome. Insulin secretion is associated with a decrease in the H2S levels. Raised H2S is detrimental in acute pancreatitis as well as in septic shock. Recently, H2S-releasing derivatives of certain drugs have shown promise in protection against gastric ulcer and in inflammatory bowel disease. The beneficial effects of certain sulphur containing herbs like ginseng and garlic may be mediated via H2S. In future, development of specific drugs modulating H2S levels may prove beneficial in varied disorders.

Therapeutic potential of herbal drugs in cerebral ischemia.

Stroke is one of the most important causes of mortality and morbidity in the world. Prevention and effective treatment of stroke is of the utmost importance. Cerebral ischemia causes disturbances in a variety of cellular and molecular mechanisms, including oxidative phosphorylation, membrane function, neurotransmitter release, and free radical generation. It has been years since tissue-type plasminogen activator (t-PA) became the first medication approved by the FDA for the management of stroke, with limited success. Thrombolytic therapy is the most effective therapeutic strategy for the prevention of brain injury and reduction of mortality in patients with cerebral infarction. However, a combination of established thrombolytic therapy and effective neuronal protection therapy may have more beneficial effects for patients with cerebral infarction. Because clinical trials of pharmacological neuroprotective strategies in stroke have been disappointing, attention has turned towards approaches which include herbal drugs that can be used in limiting the neurological damage associated with stroke. Herbal drugs may be used as prophylactic treatment in patients with high risk of stroke. Herbals drugs have been described in ancient systems of medicine for the treatment of various ailments associated with stroke and have more recently been reported to be beneficial in treating stroke. However, the strength of evidence to support the use of these herbal drugs is unclear. This review focuses on putative mechanisms underlying the beneficial effects of herbal drugs in patients with stroke and on the possibility of herbal drugs to increase the therapeutic time window in patients with cerebral ischemia.

Evaluation of antitussive activity of formulations with herbal extracts in Sulphur Dioxide (SO2) induced cough model in mice.

Cough is the most common symptom of respiratory diseases. When cough becomes serious, opioids are effective, but they have side effects like sedation, constipation, some addiction liability and also compromise the respiratory function. Therefore, there is need to have effective anti-tussive agent which do not have respiratory suppressant activity. The present study was carried out to evaluate anti-tussive activity of combination of herbal drugs as formulations in sulphur dioxide (SO2)-induced cough model in mice. Albino mice of either sex, weighing 25-30 g were divided into eight groups, (n = 6). Group 1 served as normal control, group 2 mice were given distilled water, group 3 was positive control and received codeine sulphate (10 mg/kg, p.o.) and group 4, 5, 6, 7 received coded l formulations 1, 2, 3 and 4 respectively at a dose of 0.3 ml/mice, orally, while group VIII was the vehicle control. Thirty minutes later, the mice were exposed to sulphur dioxide again for 45 sec. The mice were then placed in an observation chamber for counting of cough bouts, by two independent observers, for five minutes. All the formulations used showed significant antitussive activity in sulphur dioxide induced cough model. Thus, these formulations can prove to be useful for alleviating cough.

Evaluation of anti-inflammatory activity of plant lipids containing alpha-linolenic acid.

Two groups of fatty acids are essential to the body, the omega6 (n6) series derived from linoleic acid (18:2, n-6) and the omega3 (n3) series derived from alpha-linolenic acid (18:3, n-3). Fatty acids provide energy, are an integral part of the cell membranes and are precursors of prostaglandins, thromboxanes and leukotrienes collectively known as eicosanoids. Eicosanoids participate in development and synthesis of immunological and inflammatory responses. The fixed oils (1, 2, 3 ml/kg) containing alpha-linolenic acid, obtained from the seeds of Linseed (Linum usitatissimum), Soyabean (Glycine max) and Holy basil (Ocimum sanctum) were screened for their antiinflammatory activity using carrageenan, leukotriene and arachidonic acid induced paw edema models in rats and the antiinflammatory effects were compared with the standard drug indomethacin. Significant inhibition of paw edema was produced by all the oils in the highest dose (3 ml/kg) in all the models. While O. sanctum oil produced the maximum percentage inhibition in leukotriene induced paw edema, L. usitatissimum oil produced maximum percentage inhibition in carrageenan and arachidonic acid induced paw edema models. The results show that oils with higher alpha-linolenic acid content (L. usitatissimum and O. sanctum) produced a greater inhibition of paw edema suggesting that modulation of the course of inflammatory disorders may be achieved by altering the eicosanoid precursor (i.e. poly unsaturated fatty acids: PUFA) availability through dietary manipulation.

Protective effect of pifithrin-alpha on brain ischemic reperfusion injury induced by bilateral common carotid arteries occlusion in gerbils.

Involvement of p53 has been implicated in apoptosis induced cell death in ischemic reperfusion injury. In the present study, we have investigated neuroprotective potential of pifithrin-alpha, a p53 inhibitor in bilateral common carotid arteries occlusion (5 min) model of global cerebral ischemia in Mongolian gerbils. Gerbils were treated with pifithrin-alpha 3 mg/kg, ip. 30 min prior to occlusion. There was a significant increase in neurological symptoms and locomotor activity in ischemic animals as compared with the sham-operated animals. Increase in neurological symptoms and locomotor activity was attenuated by pifithrin-alpha 3 mg/ kg, ip. Significant increase in the number of the surviving neurons in the hippocampal CA1 pyramidal region was observed in ischemic animals treated with pifithrin-alpha 3 mg/kg, ip. This study demonstrates the neuroprotective effect of pifithrin-alpha in global cerebral ischemia in gerbils.

A reverse phase high performance liquid chromatography method for simultaneous estimation of melatonin, carbamazepine epoxide and carbamazepine simultaneously in serum.

Monitoring of plasma antiepileptic drugs is useful for better clinical management in epileptic patients, particularly in children. Carbamazepine (CBZ) is one of the commonly prescribed anticonvulsants. The active metabolite of carbamazepine-carbamazepine-10-11 epoxide (CBZ-Epo) also exhibits anticonvulsant effect. The pineal hormone, melatonin exerts an anticonvulsant effect in experimental seizure models and recently has also been used in cases of childhood epilepsy. To facilitate the simultaneous plasma estimation of carbamazepine, carbamazepine epoxide, and melatonin, a new HPLC method was developed. Waters millennium 2010 chromatography manager with a 515 HPLC pump and Waters 24879 dual absorbance UV detector was used. A 25 microlitre of sample and standards were injected, and chromatographic separation was achieved by Merck C18 reverse phase column particle size 5 micro, 250 mm x 4 mm. It was quantitated at UV light 210 nm. The retention times of melatonin, CBZ-Epo, and CBZ were 6.3 min, 7.5 min, and 13.9 min respectively. The Mobile Phase used was water: acetonitrile (70:30), pH 3.0 adjusted with orthophosphoric acid at the flow rate of 1 ml/min. The limits of detection of melatonin, carbamazepine epoxide, and carbamazepine were 800, 500, and 1300 pg respectively.

Drug-eluting stents: a pharmacoclinical perspective.

In the 26 years since Gruntzig introduced a simple balloon angioplasty technique, percutaneous coronary intervention has made extraordinary progress and has now surpassed bypass surgery in frequency. The area of coronary stenting has been the focus of intense research. One of the major problems encountered after stenting is an exaggerated vascular neointimal proliferation called in-stent stenosis. The evolution of drug-eluting stents has helped in reducing the incidence of in-stent stenosis by almost half. A number of pharmacological agents have been tried in coronary stents with varying degrees of success; many more are being developed and tested. Serious doubts have been expressed about the pharmacoeconomics of drug-eluting stents compared with bare metal stents, because of the huge disparity in costs. Drug-eluting stents, which can be grouped under both device and instrument, have thrown up interesting challenges for clinical trials. The future could see the development of more compact devices with the help of diverse fields such as nanotechnology, microelectronics and advanced materials technology.

Contribution of genomics and proteomics in understanding the role of modifying factors in Parkinson's disease.

Parkinson's disease (PD) is a complex neurological disorder, characterized by selective degeneration of nigrostriatal dopaminergic neurons. It is a multi-factorial disease, contributed by a combination of age, genetic and environmental factors. Etiology of sporadic PD and mechanism underlying selective loss of dopaminergic neurons has not yet been clearly understood. Recent developments in genomics and proteomics have revolutionized the research on PD at genetic level. Differential gene expression patterns (DNA biochip technology), age-dependent complex genetic patterns (SNP genotyping), and protein expression profiles (proteomics) of PD patients have started providing the specific and rigorous molecular explanation and role of modifying factors in PD. Genomics and proteomics are further expected to help in developing biomarkers for diagnosis of early onset PD and also to develop valuable and potential therapeutic strategies for its treatment. In this review, we have discussed the progress made by genomics and proteomics, in understanding the role of modifying factors in PD.

Protective effect of endothelin antagonist (TAK-044) on neuronal cell viability in in vitro oxygen-glucose deprivation model of stroke.

The present study was carried to investigate the effect of endothelin antagonist (TAK-044) in an in vitro model of stroke using primary neuronal culture. Hypoxia in neuronal culture was induced for 3 h using oxygen glucose deprivation (OGD) model, thereafter cells were reperfused. In separate group cultures were incubated with graded concentrations of TAK-044 (0.01, 0.1 and 1 microg/microl) for different time duration i.e. 6, 12 and 24 hours after reperfusion. Percent cell viability was assessed 24 h after reperfusion using MTT assay. It was observed that percent cell viability was reduced to 13.7 +/- 0.4% in the cells under 3 h hypoxic condition as compared to the cells under normal condition (100%). TAK-044 at the concentrations of 0.1 and 1 microg/microl, but not at 0.01 microg/microl showed significant (P<0.01) improvement in the % cell viability as compared to the cells in hypoxic condition. Percent cell viability at the concentration of 0.1 and 1 microg/microl for 24 h time duration after reperfusion were 54.8 +/- 3.2% and 75.4 +/- 1.8% respectively as compared to the cells under hypoxic condition (13.7 +/- 0.4%). The results demonstrate the neuroprotective effect of TAK-044 against neuronal damage caused by hypoxia induced in neuronal culture.

Post traumatic epilepsy: a review of scientific evidence.

Post traumatic epilepsy is the development of recurrent seizures following head trauma and has a high clinical relevance. Several risk factors including some genetic factors increase the susceptibility of post traumatic epilepsy. The precise mechanisms of epileptogenesis in post-traumatic epilepsy are still poorly understood. Many structural, physiologic and biochemical changes in the brain may account for epileptogenesis. The reactive oxygen species (ROS), especially *OH and excitotoxicity are primarily involved. Antioxidants, like tocopherol, antiepileptic drug zonisamide, condensed tannins, melatonin, adenosine, trans-resveratrol, and some other agents have been proposed to prevent epileptogenic focus formation. The review also discusses various aspects of post traumatic epilepsy, mechanisms of epileptogenesis, and clinical implications.

Modulation of serum concentrations of melatonin by carbamazepine and valproate.

Evidence has accumulated about the involvement of reactive oxygen species (ROS) in epilepsy. The neuromodulator melatonin has been shown to reduce oxidative stress in various animal models due to its free radical scavenging properties. The present study investigated whether carbamazepine and valproate alter serum concentrations of melatonin. Epileptic children were randomly assigned to receive carbamazepine/ valproate monotherapy till 22 patients were recruited in the study. At the tenth day, in the evening, samples were drawn for baseline endogenous melatonin estimation. The patients were then administered exogenous melatonin, and repeat samples were drawn after 30 minutes. Serum levels of melatonin were estimated using Melatonin ELISA kits. The median levels of melatonin were 165.0 pg/ml (Range 50.0-350.0) in CBZ+MEL group and 78.0 pg/ml (Range 13.0-260.0) in the VPA+MEL group. The observed difference in melatonin levels could be attributed to the difference in antiepileptic drugs, additive increase in reactive oxygen species due to disease combined with carbamazepine, or possibly to a difference in melatonin kinetics in conditions of oxidative stress.

Effect of alpha lipoic acid, melatonin and trans resveratrol on intracerebroventricular streptozotocin induced spatial memory deficit in rats.

In the present study, the effect of antioxidants-alpha lipoic acid, melatonin and trans resveratrol were studied against intracerebroventricular streptozotocin induced spatial memory deficit. Male Wistar rats were injected with intracerebroventricular streptozotocin bilaterally. The rats were treated chronically with alpha lipoic acid (200 mg/kg, po), melatonin (20 mg/kg, ip) and trans resveratrol (20 mg/kg, ip) for 18 days starting from day 1 of streptozotocin injection in separate groups. The spatial memory was evaluated using the Morris water maze task. The intracerebroventricular streptozotocin rats treated with antioxidants showed significantly less spatial memory deficit both in the acquisition and probe trials as compared to the vehicle treated rats. The study demonstrated the effectiveness of alpha lipoic acid, melatonin and trans resveratrol in preventing spatial memory deficit induced by intracerebroventricular streptozotocin and it's potential in age related neurodegenerative disorders where oxidative stress is involved such as Alzheimer's disease.

Contribution of genomics, proteomics, and single-nucleotide polymorphism in toxicology research and Indian scenario.

Advancement in the molecular tools used in toxicology has provided immense information about the cellular and global structure and function of toxicant-responsive genes. Now, it has become possible to assess the functional activity of genes and proteins involved in various toxicological pathways, which were not possible with the conventional methods. Many genes are known to have a greater influence on the susceptibility to environmental agents than others; therefore, identification and characterization of polymorphism in such genes for the determination of early, late, or no response of an individual for the toxicant-induced diseases has also become mandatory. Toxicogenomics, a newly born discipline of toxicology, comprises of two major facets, one, how various genes in the genome respond to environmental toxicants and stressors and second, how toxicants modify the function and expression of specific genes in the genome. Toxicogenomics play an important role in the identification and characterization of molecular biomarkers to predict cellular toxicity and to determine the efficacy and exposure in the toxicity trials at an early stage. Genome and proteome-wide expression profiles in combination with conventional toxicology are being used to classify compounds, predict the mechanism of toxicity of newer compounds and determine the susceptibility of an individual for the toxic responses. Single-nucleotide polymorphism in toxicant-responsive genes is being used to obtain basic information of the genetic variation and its role in the functional protein expression. Various national and international government and private organizations have launched several programs on gene-environment interactions. Council of Scientific and Industrial Research (CSIR), New Delhi, India, has also launched a program on 'toxicogenomics of genetic polymorphism in Indian population to industrial chemicals for development of biomarkers' to provide better ventures and facilities to researchers in order to understand the environment-genome interactions. In this review, the contribution of genomics, proteomics, and SNPs in toxicology along with its current status in India has been discussed3.

Expression of opioid receptor-like 1 (ORL1) & mu opioid receptors in the spinal cord of morphine tolerant mice.

BACKGROUND & OBJECTIVE: The mechanism underlying the development of tolerance to morphine is not clearly understood though a number of factors have been implicated. One of the likely factors may be increased activity of anti-opioid peptides like nociceptin (also known as orphanin FQ or N/OFQ). N/OFQ and morphine bind to opioid receptor-like 1 (ORL1) receptor and muopioid receptor respectively. The present work was undertaken to investigate the density of ORL1 and mu (mu) receptor expression in the spinal cord of mice after inducing morphine tolerance. METHODS: Swiss albino mice were injected with either morphine (experimental group, n=15) or saline (control, n=15), twice a day for 9 days. The development of tolerance was noted by the hotplate test. Cryostat sections of the cervical region of spinal cord were labeled with specific ligands to localize ORL1 and mu receptors. The density of receptor expression over laminae I-II of spinal cord was evaluated using image analysis system. RESULTS: The morphine treated mice developed tolerance by day 9 as evident by the hot plate test. Both receptors were selectively expressed at a higher concentration over the superficial laminae (I-II) of the dorsal horn, indicating a role in pain processing. An increased expression of ORL1 receptors was also noted over the gray matter around the central canal. Quantitative analysis showed an increased expression of ORL1 and mu receptors though the increase was not statistically significant. INTERPRETATION & CONCLUSION: The present study showed that both, ORL1 and mu-opioid receptors were expressed in areas of the spinal cord, concerned with transmission of pain signals. The density of these receptors increased in the superficial laminae (I-II) though not significantly from control after morphine tolerance. The increase in ORL1 receptors could oppose the analgesic action of morphine, contributing to tolerance. Further studies need to be done to elucidate the mechanism of morphine tolerance.

Animal models of cerebral ischemia for evaluation of drugs.

Stroke is a major cause of death and disability worldwide. The resulting burden on the society continues to grow, with increase in the incidence of stroke. Brain attack is a term introduced to describe the acute presentation of stroke, which emphasizes the need for urgent action to remedy the situation. Though a large number of therapeutic agents like thrombolytics, NMDA receptor antagonists, calcium channel blockers and antioxidants, have been used or being evaluated, there remains a large gap between the benefits by these agents and properties an ideal drug for stroke should offer. In recent years much attention is being paid towards the exploration of herbal preparation, antioxidant agents and combination therapies including COX-2 inhibitors in experimental model of stroke. For better evaluation of the drugs and enhancement of their predictability from animal experimentation to clinical settings, it has been realized that the selection of animal models, the parameters to be evaluated should be critically assessed. Focal and global cerebral ischemia represents diseases that are common in the human population. Understanding the mechanisms of injury and neuroprotection in these diseases is important to learn new target sites to treat ischemia. There are many animal models available to investigate injury mechanisms and neuroprotective strategies. In this article we attempted to summarize commonly explored animal models of focal and global cerebral ischemia and evaluate their advantages and limitations.