Immunohistochemical detection of 8-hydroxydeoxyguanosine: A biomarker of oxidative DNA damage in oral submucous fibrosis.

BACKGROUND: Oral submucous fibrosis (OSMF) is one of the common potentially malignant disorders prevailing in India. The primary etiological factors include tobacco and arecanut, which contain numerous reactive oxygen species (ROS). ROS attack guanine bases in DNA and form 8-hydroxydeoxyguanosine (8-OHdG), which can be detected in patients who have diseases associated with oxidative stress. The oxidative DNA damage produced by oxidative stress may induce malignant transformation. AIM: The aim of the present study is to detect the expression of 8-OHdG in OSMF patients and compare the expression within different grades of OSMF and also normal buccal mucosa. MATERIALS AND METHODS: A total of 30 samples were examined for the immunohistochemical expression of 8-OHdG. The control group included 10 formalin-fixed paraffin-embedded tissue blocks of the normal buccal mucosa. The study group includes 20 cases of formalin-fixed paraffin-embedded tissue blocks of OSMF (5 cases in each grade of very early, early, moderately advanced and advanced cases of OSMF). Three-micron thick tissue sections were made from each sample and stained with 8-OHdG antibody. The results were statistically analyzed using Kruskal-Wallis and Mann-Whitney U test. RESULTS: Statistically significant difference exists in the intensity of 8-OHdG expression between the study groups. The P-value obtained was <0.001, which was highly statistically significant. CONCLUSION: The present study is the first attempt to evaluate the expression of 8-OHdG in tissue samples of OSMF that revealed the role of free radicals and oxidative DNA damage in these patients. Further research with larger sample size, clinicopathologic correlation and long-term follow-up will shed more light on the pathogenesis of OSMF. It will also be useful for the development of new therapeutic strategies targeting treatment modalities for OSMF.

Therapeutic investigations of novel indoxyl-based indolines: A drug target validation and Structure-Activity Relationship of angiotensin-converting enzyme inhibitors with cardiovascular regulation and thrombolytic potential.

A family of 12 members of Naphthalene-2-ol-indolin-2-one-thiocarbamides (5a-l) with pharmacological potentials of cardiovascular modulator were efficiently synthesized and evaluated. These compounds show inhibitory activity on angiotensin-converting enzyme (ACE), which is a principal constituent of the renin-angiotensin system and causative source for hypertension (HTN) (elevated blood pressure) and congestive heart failure (CHF), a parameter that was tested in this report. Prior to this, to get more insight into the binding mode and inhibition of human ACE C-domain (PDB ID: 2XY9) and N-domain (PDB ID: 3NXQ) compounds 5a-l was docked into the active site of them. The established inhibitory constant (Ki) (range 40-500 nM) and least binding affinities (-18.52 to -30.57 kcal/mol) indicated the therapeutic selectivity of compounds 5a-l towards ACE C-domain inhibition over ACE N-domain. The cytotoxicity effect of most potent compounds among 5a-l were tested in normal breast cells and MCF-7 cell lines. Simultaneously, H2O2 induced antioxidant and DNA damage assessment was executed. Eventually, a thrombolytic activity followed by a human red blood cell (HRBC) membrane stabilization study to ensure the relaxation of blood and stabilization of RBC was executed. Structure-Activity Relationship (SAR) study discloses the potential of 5c, 5h, and 5k as cardiovascular protective therapeutic agents among 5a-l.

Biosynthesis of gold nanoparticles and related cytotoxicity evaluation using A549 cells.

Biosynthesis of gold nanoparticles (AuNPs) has become an attractive area of research as it is environmentally benign. The toxicity of AuNPs synthesized by chemical routes has been widely studied. However, little is known about the toxicity associated with the biological synthesis of AuNPs. The present study was carried out to synthesize AuNPs using star anise (Illicium verum; a commercially available spice in abundance)and evaluate its toxicity using human epithelial lung cells (A549) in comparison with AuNPs synthesized by the traditional chemical methods (using sodium citrate and sodium borohydride). Apart from cell viability, markers of oxidative stress (reduced glutathione) and cell death (caspases) were also evaluated to understand the mechanisms of toxicity. Cell viability was observed to be 65.7 percent and 72.3 percent in cells exposed to chemically synthesized AuNPs at the highest dose (200nM) as compared to 80.2 percent for biologically synthesized AuNPs. Protective coating/capping of AuNPs by various polyphenolic compounds present in star anise extract appears to be a major contributor to lower toxicity observed in biologically synthesized AuNPs.

DNA/protein interaction and cytotoxic activity of imidazole terpyridine derived Cu(II)/Zn(II) metal complexes.

Two imidazole terpyridine (itpy) based complexes, [Cu(itpy)(OAc)(H2O)]NO3·H2O (1) and [Zn(itpy)(OAc)]OAc (2) have been synthesised and characterized. The crystal structure of complex 1 shows distorted octahedral geometry with an anti-parallel stacking arrangement. The interactions of the two complexes with Calf thymus DNA (ctDNA) have been studied using absorption titration and circular dichroism. Complex 1 shows coordinate binding to DNA bases, and complex 2 shows an intercalative mode of binding with DNA. Complex 1 cleaves the DNA via an oxidative pathway in the presence of additives, because of the presence of a redox active copper(II) centre. However, complex 2 cleaves DNA hydrolytically. Interactions of the two complexes with bovine serum albumin have been studied using fluorescence quenching and circular dichroism experiments. Circular dichroic analysis reveals that both the complexes strongly influence the secondary structure of the protein. Fluorescence quenching experiments indicate that there are different binding sites for complexes 1 and 2 on the protein. Furthermore, the complexes show potential cytotoxicity towards the A549 lung cancer cell line. Both the complexes have been found to induce apoptosis.

Non-hypotensive dose of telmisartan and nimodipine produced synergistic neuroprotective effect in cerebral ischemic model by attenuating brain cytokine levels.

The hypothesis of the present study is that the anti-inflammatory property of telmisartan (TM), an AT1 blocker that may exert neuroprotection through attenuation of excitatory amino acids by controlling cytokines and reactive oxygen species, release during ischemia. The neuroprotective effect of TM and its combination with nimodipine (NM) were studied in rats by using middle cerebral artery occlusion method followed by ischemic reperfusion (IR) after 2 h of occlusion. The drugs were administered 30 min prior to the surgery and continued throughout the study period. After 24 h of IR the neurological deficit was assessed, and the locomotor activity and open field behaviour were assessed on the seventh day. On the ninth day, the brains were isolated for neurochemical and cytokine measurements and histopathological studies. The results have shown that treatment of TM (5 & 10 mg/kg) gradually reduced the glutamate, aspartate and glutamine synthetase levels. It also restored the ATP, Na(+)K(+)ATPase, glutathione and synapse integrity in the different regions of the brain in comparison to ischemic brain. TM ameliorated the pro-inflammatory cytokine (IL-1ß, IL-6, TNF-α), lipid peroxide and nitric oxide levels. Anti-inflammatory cytokine IL-10 level was found to be concurrently increased. Combination therapy of TM with NM (5 mg/kg) has shown additive effects in the above said parameters. Further a positive correlation between glutamate and cytokine release was observed, and it indicated that synaptic clearance of glutamate can be regulated by cytokines. It can be concluded that TM induces neuroprotective activity through amelioration of pro-inflammatory cytokine release during cerebral ischemia. The additive effect of NM on TM neuroprotective effect would be through controlling cytokine release, ATP restoration by cerebrovasodilation, and along with prevention of Ca(2+) dependent glutamate toxicity in neurons. The advantage of TM therapy in ischemic state can be explored clinically due to its dual effect in hypertension.

Immobilization of flavin adenine dinucleotide (FAD) onto carbon cloth and its application as working electrode in an electroenzymatic bioreactor.

A high porosity carbon cloth with immobilized FAD was employed as working electrode in electrochemical NADH-regeneration procedure. Carbon cloth was oxidized with hot acids to create surface carboxyl group and then coupled by adenine amino group of FAD with carbodiimide in the presence of N-hydroxysulfosuccinimide. The bioelectrocatalytic NADH-regeneration was coupled to the conversion of achiral substrate pyruvate into chiral product l-lactate by l-lactate dehydrogenase (l-LDH) within the same reactor. The conversion was completed at 96h in bioreactor with FAD-modified carbon cloth, resulting in about 6mM of l-lactate from 10mM of pyruvate. While with bare carbon cloth, the yield at 120h was around 5mM. Immobilized FAD on the surface of carbon cloth electrode facilitated it to carry electrons from electrode to electron transfer enzymes; thereby NADH-regeneration was accelerated to drive the enzymatic reaction efficiently.

1-Meth-oxy-4-({[(4-meth-oxy-phen-yl)-sulfan-yl](phen-yl)meth-yl}sulfan-yl)benzene.

The title compound, C(21)H(20)O(2)S(2), forms a propeller-shaped structure with the tetra-hedral C atom as the central hub and meth-oxy-benzene and phenyl residues as radiating blades. Short C-H⋯π contacts are observed.

Removal of a potent cyanobacterial hepatotoxin by peat.

Microcystins (cyclic heptapeptides), produced by a number of freshwater cyanobacteria, are of health concern in potable water supplies. In this article, the adsorptive removal of microcystin-RR (MCRR) from the aqueous solution by a low-cost adsorbent, peat, was investigated. The BET surface area of peat was found to be 12.134 m2/g. The adsorption process was pH dependent, with maximum adsorption occurring at pH 3. Kinetic studies revealed that the adsorption of MCRR onto peat was a rapid process. The adsorption capacity (Qmax) as revealed by the Langmuir model was found to be 286 µg/g at pH 3. Among various desorption media studied, strong alkali solutions (2 N NaOH) showed the highest desorption (97%). Thus, peat has potential to be used as an adsorbent for the removal of the cyanotoxin, MCRR, from drinking water.

Experimental studies on removal of microcystin-LR by peat.

Cyanotoxins have caused worldwide concerns for their eclectic occurrence and toxic effects, which led to an intensive search of cost-effective techniques for their removal from contaminated waters. A range of biomaterials was tested for their efficacy to adsorb a potent cyanotoxin, microcystin-LR (MCLR). Among these sorbents, peat showed the maximum efficacy to sequester MCLR. The BET (Brunauer-Emmett-Teller) surface area of peat was found to be 12.134 m(2)/g. The pH of the reaction media played a significant role in removal of MCLR; maximum adsorption occurred at pH 3. Kinetic studies showed that the adsorption of MCLR onto peat was a rapid process. The adsorption capacity (Q(max)) from the Langmuir model was found to be 255.7 µg/g at pH 3. Among various desorption media studied, strong alkali (2N NaOH) showed highest desorption (94%).

Role of bicontinuous microemulsion in the rapid enzymatic hydrolysis of (R,S)-ketoprofen ethyl ester in a micro-reactor.

Bicontinuous microemulsion was employed as the medium for enzymatic hydrolysis of (R,S)-ketoprofen ethyl ester in the presence of esterase for the first time. In addition, a methodology for the separation of optically pure ketoprofen from the microemulsion system for analysis by gas chromatography was developed. Various factors influencing the enzymatic hydrolysis of (R,S)-ketoprofen ethyl ester such as temperature, enzyme concentration and reaction time were optimized experimentally. The enzymatic hydrolysis in a bicontinuous microemulsion system showed a final conversion of 84.6% after 50 h of reaction, while hydrolysis in Tris-HCl buffer solution resulted in only 26.9% conversion after 150 h without completing the reaction. A comparison of the rate of the enzymatic hydrolysis reaction with rates of reaction in other biphasic media revealed that the bicontinuous microemulsion system was faster and more advantageous. The extremely large interfacial area of the latter fluid likely facilitated the contact between the catalyst and the substrate. Because the enzyme applied was not selective, formation of (R)-ketoprofen was also observed. Therefore, application of an enzyme with higher selectivity would provide better results.

Biocrystallization of silver and gold ions by inactive cell filtrate of Rhizopus stolonifer.

Microbes and their cell filtrates are known to synthesize metal nanoparticles. But maintenance of aseptic conditions and irregularly shaped and sized nanoparticles are major drawbacks of the system. In this study cell filtrate from inactive biomass of Rhizopus stolonifer was used for the first time to produce near uniformly sized and shaped Ag and Au nanoparticles at room temperature. The size of Ag and Au nanoparticles were found in the range of 25-30 nm and 1-5 nm, respectively. UV-vis spectrum, TEM and XRD measurements confirmed the formation of Ag and Au nanoparticles.

Bioreduction of trivalent aurum to nano-crystalline gold particles by active and inactive cells and cell-free extract of Aspergillus oryzae var. viridis.

Bioreduction efficacy of both active (AB) and inactive (IB) cells/biomass of Aspergillus oryzae var. viridis and their respective cell-free extracts (ACE and ICE) to convert trivalent aurum to gold nanoparticles were tested in the present study. Strong plasmon resonance of gold nanoparticles was observed between 540 and 560 nm in the samples obtained from AB, IB, ACE and ICE. Transmission electron microscopy (TEM), field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray (EDX) and X-ray diffraction (XRD) were performed to examine the formation of gold nanoparticles. Comparing all four forms of A. oryzae var. viridis, ICE showed high gold nanoparticle productivity. The nanoparticles formed were quite uniform in shape and ranged in size from 10 to 60 nm. In addition some triangle, pentagon and hexagon-shaped nanoplates with size range of 30-400 nm were also synthesized especially at lower pH. Organics from the inactive cells are believed to be responsible for reduction of trivalent aurum to nano-sized gold particles. Organic content of the ICE was found to be double the amount of ACE. High productivity of gold nanoparticles by metabolic-independent process opens up an interesting area of nanoparticle synthesis using waste fungal biomass from industries.

Platinum recovery from ICP wastewater by a combined method of biosorption and incineration.

A high performance biosorbent, polyethylenimine (PEI)-modified biomass, was prepared by attaching PEI onto the surface of inactive Escherichia coli biomass. Wastewater containing platinum was collected from an industrial laboratory for inductively coupled plasma (ICP) and used for the recovery study. The maximum platinum uptake of PEI-modified biomass was enhanced up to 108.8 mg/g compared to 21.4 mg/g of the raw biomass. Kinetic experiments revealed that sorption equilibrium could reach within 60 min for the PEI-modified biomass. The results of FTIR and XPS analysis of Pt-unloaded and Pt-loaded PEI-modified biomass indicated that electrostatic interaction was the main binding mechanism between the platinum ions and the binding sites on the surface of the biomass. Metallic form of platinum in ash was recovered by incineration with a recovery efficiency of over 98.7%. Furthermore, XPS, TEM and XRD results confirmed that the platinum recovered were both forms of Pt(0) and Pt(2+).

Nitrate removal using Brevundimonas diminuta MTCC 8486 from ground water.

Brevundimonas diminuta MTCC 8486, isolated from marine soil of coastal area of Trivandrum, Kerala, was used for biological removal of nitrate from ground water collected from Kar village of Pali district, Rajasthan. The organism was found to be resistance for nitrate up to 10,000 mg L(-1). The optimum growth conditions for biological removal of nitrate were established in batch culture. The effect of carbon sources on nitrate removal was investigated using mineral salt medium (MSM) containing 500 mg L(-1) of nitrate to select the most effective carbon source. Among glucose and starch as carbon source, glucose at 1% concentration increased the growth (182+/-8.24 x 10(4) CFU mL(-1)) and induced maximum nitrate reduction (86.4%) at 72 h. The ground water collected from Kar village, Pali district of Rajasthan containing 460+/-5.92 mg L(-1) of nitrate was subjected to three different treatment processes in pilot scale (T1 to T3). Higher removal of nitrate was observed in T2 process (88%) supplemented with 1% glucose. The system was scaled up to 10 L pilot scale treatment plant. At 72 h the nitrate removal was observed to be 95% in pilot scale plant. The residual nitrate level (23+/-0.41 mg L(-1)) in pilot scale treatment process was found to be below the permissible limit of WHO.

Phyto-crystallization of palladium through reduction process using Cinnamom zeylanicum bark extract.

In this paper we studied the potential of nanocrystalline palladium particle production using Cinnamom zeylanicum bark extract (CBE) as the biomaterial for the first time. We studied the effects of biomaterial dosage, pH and temperature on nanoparticle formation; none of these factors had a major effect on the size and shape of the nanoparticles formed. Transmission electron microscopy (TEM) observations confirmed the synthesis of nano-sized palladium particles. More or less uniformly sized palladium nanoparticles were synthesized with an average size ranging from 15 to 20 nm. It was found that the zeta potential of these formed palladium nanoparticles was negative, and that it increased with an increase in pH. Energy dispersive X-ray (EDX) analysis results confirmed the significant presence of palladium. Of the palladium ions, 60% were reduced to a zero valent form by CBE. Terpenoids are believed to play an important role in palladium nanoparticle biosynthesis through the reduction of palladium ions. Currently, however, the exact mechanism for the synthesis of palladium nanoparticles is unclear. Our protocol for the phyto-synthesis of palladium nanoparticles under moderate pH and room temperature offers a new means to develop environmentally benign nanoparticles.

Cinnamon zeylanicum bark extract and powder mediated green synthesis of nano-crystalline silver particles and its bactericidal activity.

The exploitation of various plant materials for the biosynthesis of nanoparticles is considered a green technology as it does not involve any harmful chemicals. The present study reports the synthesis of silver (Ag) nanoparticles from silver precursor using the bark extract and powder of novel Cinnamon zeylanicum. Water-soluble organics present in the plant materials were mainly responsible for the reduction of silver ions to nano-sized Ag particles. TEM and XRD results confirmed the presence of nano-crystalline Ag particles. The pH played a major role in size control of the particles. Bark extract produced more Ag nanoparticles than the powder did, which was attributed to the large availability of the reducing agents in the extract. Zeta potential studies showed that the surface charge of the formed nanoparticles was highly negative. The EC(50) value of the synthesized nanoparticles against Escherichia coli BL-21 strain was 11+/-1.72 mg/L. Thus C. zeylanicum bark extract and powder are a good bio-resource/biomaterial for the synthesis of Ag nanoparticles with antimicrobial activity.

Hepatoprotective and curative properties of Kombucha tea against carbon tetrachloride-induced toxicity.

Kombucha tea (KT) is sugared black tea fermented with a symbiotic culture of acetic acid bacteria and yeasts, which is said to be tea fungus. KT is claimed to have various beneficial effects on human health, but there is very little scientific evidence available in the literature. In the present study, KT along with black tea (BT) and black tea manufactured with tea fungus enzymes (enzyme-processed tea, ET) was evaluated for hepatoprotective and curative properties against CCl4-induced toxicity, using male albino rats as an experimental model by analyzing aspartate transaminase, alanine transaminase, and alkaline phosphatase in plasma and malondialdehyde content in plasma and liver tissues. Histopathological analysis of liver tissue was also included. Results showed that BT, ET, and KT have the potential to revert the CCl4-induced hepatotoxicity. Among the three types of teas tried, KT was found to be more efficient than BT and ET. Antioxidant molecules produced during the fermentation period could be the reason for the efficient hepatoprotective and curative properties of KT against CCI4-induced hepatotoxicity.

Antibacterial activity of hydroxyalkenyl salicylic acids from sarcotesta of Ginkgo biloba against vancomycin-resistant Enterococcus.

A chloroform fraction prepared from the sarcotesta of Ginkgo biloba showed potent inhibitory activity against vancomycin-resistant Enterococcus (VRE). The active compounds were elucidated to be 2-hydroxy-6-(8-pentadecenyl) salicylic acid (1) and 2-hydroxy-6-(10-heptadecenyl) salicylic acid (2) based on their spectral analysis. Compounds 1 and 2 showed significant antibacterial activities against VRE.

Bio-separation of toxic arsenate ions from dilute solutions by native and pretreated biomass of Aspergillus fumigatus in batch and column mode: effect of biomass pretreatment.

The sorption of As(V) from aqueous solution onto live and pretreated biomass of Aspergillus fumigatus was studied. The studies on optimization of contact time, adsorbent dosage and pH showed that the FeCl(3) treated and FeSO(4) treated biomass had the maximum capacity to adsorb As(V) while acid treated biomass was found to be minimum. Adsorption parameters were determined using both Langmuir and Freundlich isotherm models. The maximum adsorption capacity of 0.054 mg/g was observed in FeCl(3) treated and FeSO(4) treated biomasses. Column mode studies were conducted using FeSO(4) treated biomass to compare its efficacy with batch mode to adsorb As(V). Batch mode experiments proved to be efficient. Desorption studies were also carried out with dilute sodium hydroxide to recover both the adsorbent and adsorbate.