Purification and Characterization of Novel Antihypertensive and Antioxidative Peptides From Whey Protein Fermentate: In Vitro, In Silico, and Molecular Interactions Studies.

OBJECTIVE: The goal of this research was to purify and characterize the novel angiotensin-converting enzyme (ACE)-inhibitory and antioxidant peptides from fermented whey protein concentrate produced by Lactobacillus paracasei and Saccharomyces cerevisiae in a co-fermentation system. METHOD: Whey protein fermented with lactic acid bacteria and yeast culture was analyzed for antioxidative, ACE inhibition, as well as anti-inflammatory activity followed by SDS-PAGE, isoelectric focusing, and 2-dimensional (2D) analysis. Anti-inflammatory activity of whey protein fermentate was also studied on the RAW 264.7 cell line. The bioactive peptides were separated from the whey protein fermentate using reverse-phase high-performance liquid chromatography (RP-HPLC) and reverse-phase liquid chromatography mass spectrometry (RPLC/MS), and thus identification and characterization of purified bioactive peptide was performed. RESULTS: Whey protein fermentate samples' bioactivity was analyzed at specific time intervals at 12, 24, 36, and 48 hours at 37 °C for M11 and at 25 °C for WBS2A. The development settings (incubation time [12, 24, 36, and 48 hours) and inoculation rates [1.5%, 2.0%, and 2.5%]) were optimized for peptide synthesis via the o-phthaldialdehyde (OPA) method (proteolytic activity). Maximum proteolytic activity was observed at 37 °C for M11 (6.50 mg/mL) and at 25 °C for WBS2A (8.59 mg/mL) for 48 hours of incubation. Protein profiling was carried out using SDS-PAGE and 2D gel electrophoresis, in which Sodium dodecyl-sulfate (SDS) exhibited protein bands in the 10- to 55-kDa range, while 2D showed protein bands varying from 10 to 70 kDa. Every spot from 2D was digested by trypsin and identified by RPLC/MS. Protein fractionations (3- and 10-kDa permeates) were carried out employing RP-HPLC. Whey protein fermentate has anti-inflammatory action in RAW 264.7 macrophages that have been exposed to lipopolysaccharide. A molecular docking system was also used to investigate the interactions of peptides (AFLDSRTR, ILGAFIQIITFR) with human myeloperoxidase enzyme. CONCLUSIONS: The antihypertensive and antioxidative peptides discovered from whey protein fermentate may be helpful in the design of pharmacologically active healthy ingredients in the upcoming years.

Association of PPARγ gene expression with postprandial hypertriglyceridaemia and risk of type 2 diabetes mellitus.

PURPOSE: Peroxisome proliferator-activated receptor γ (PPARγ) gene is strongly associated with type 2 diabetes mellitus, as well as postprandial lipemia, and plays an important role in Wnt dependent adipogenesis in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT). We aimed to study the expression of PPARγ gene in SAT and VAT to find out its correlation with postprandial hypertriglyceredemia and glucose intolerance. METHODS: Thirty subjects who were scheduled to undergo abdominal surgery were recruited in three groups (n = 10 in NGT, n = 10 in prediabetes, and n = 10 in T2DM). A standardized oral fat challenge was performed. Anthropometry, plasma glucose, HbA1c, and fasting serum insulin were also measured. SAT and VATs were collected during surgery for PPARγ gene expression studies by real-time PCR. RESULTS: PPARγ gene expression was 5.5-fold lower in T2DM and 1.7-fold lower in prediabetes as compared with NGT subjects in VAT. There was a significant negative correlation of expression of PPARγ gene in VAT {Tgauc (r = -0.57, p < 0.007), Peak Tg (r = -0.51, p < 0.01)} as well as in subcutaneous adipose tissue {Tgauc (r = -0.45, p < 0.02)} with PPTg responses measures. CONCLUSION: Reduced adipocyte expression of PPARγ gene and the resultant postprandial hypertriglyceredemia is associated with greater risk of diabetes and prediabetes.

Understanding the coagulant activity of zirconium oxychloride to control THMs formation using response surface methodology.

In the present study, impact of coagulant activity of zirconium oxychloride and aluminium sulphate on the kinetics of chlorine consumption and trihalomethanes (THMs) formation has been delineated. Zirconium Oxychloride showed rapid chlorine decay within the first 30 min, which further achieved steady rate after 60 min, but in case of aluminium sulphate chlorine consumption has been increased drastically throughout the chlorine decay. Zirconium oxychloride has effectively reduced significant amount of slow reducing agents (SRA) as well as fast reducing agents (FRA), which correspond to the rate of reduction in phenolic groups from water enriched with Natural Organic Matter (NOM) which eventually decreased trihalomethane mediated cancer risk by ~ 2.3 times among adults as compared to aluminium sulphate. Result depicts the outstanding coagulant activity of zirconium oxychloride as it tends to surpass aluminium sulphate in reducing NOM "measured as Absorbance Slope Index (ASI)" and phenol by 57.98% and 49.02% respectively from NOM enriched chlorinated water, which also resembles the THMs removal trend observed during cancer risk assessment.

Comparative study of anatomical specimens using plastination by araldite HY103, polypropylene resin, 6170H19 Orthocryl and silicone - A qualitative study.

BACKGROUND: Most of the organs and tissues are preserved in formalin with its own set of disadvantages. Plastination is a unique method of permanently preserving tissue in a life like state. Plastination developed by western authorities is a labour and equipment intensive affair. Most common polymer used is S10, however this study uses easily available alternative polymers for plastination. METHOD: Various polymers like Epoxy resins, Polypropylene resins, Orthocryl and silicone were used in plastinating the anatomical specimens. Specific methods were used for solid, hollow organs and brain specimens. The specimens were made to undergo stages of fixation, dehydration, impregnation and curing. The results were studied and interpreted under various parameters. RESULTS: The results were interpreted under various parameters like shrinkage, retention of colour, odour, pliability and retention of gross anatomy. The study concluded that Orthocryl and Epoxy resins retained maximum colour with minimal shrinkage while maximum discolouration was with polypropylene plastinates. Brain sections were best preserved in Orthocryl. CONCLUSION: The study concluded that indigenous methods and materials can produce quality plastinates which can be an important adjunct to traditional methods of teaching however more studies need to be done for refinement.

Multi-exposure cancer and non-cancer risk assessment of trihalomethanes in drinking water supplies - A case study of Eastern region of India.

The lifetime cancer risk and the hazard index of trihalomethanes (THMs) through oral ingestion, dermal absorption, and inhalation exposure from supply water of five WTPs were analysed. THMs concentration varied from plant to plant and was found to be in the range of 274-511µg/l, which is much higher than the prescribed USEPA standards of 80µg/l. Chloroform was the most dominant THM followed by bromodichloromethane (BDCM), and dibromochloromethane (DBCM). Cancer risk analysis through multi-pathways exposure reveals that residents had a higher cancer risk through oral ingestion than other two routes of exposure. The lifetime cancer risks of THMs from supply water were 100 times higher than prescribed USEPA guidelines. The higher cancer risk found for Indian context than those reported for other countries like USA, UK, Japan, Australia, is mainly due to the higher concentration level of THMs, water intake and average body weight. The study also revealed that amongst different THMs, chloroform is the major THMs causing cancer risk through both oral and dermal route of exposure whereas in case of inhalation it was mainly because of BDCM. Average lifetime cancer risk analysis indicated that females are more prone to cancer risk than males. Oral ingestion is a major route indicating the potential impact of non-cancer risk while it was insignificant through dermal exposure. Sensitivity analysis of THMs revealed that chloroform is the predominant parameter followed by body weight and exposure duration influencing cancer risk.

Extraction of nickel by microbial reduction of lateritic chromite overburden of Sukinda, India.

Microbial extraction of nickel from lateritic chromite overburden (COB), Sukinda by Acidithiobacillus ferrooxidans has been investigated in this work. In anoxic environment, A. ferrooxidans reduced the ferric iron in goethite [Fe(O)OH] mineral of COB by using elemental sulphur as electron donor. Nickel embedded in the complex goethite matrix of COB was successfully recovered by cumulative action of sulphuric acid, generated by oxidation of elemental sulphur and reduction of ferric iron in goethite matrix by A. ferrooxidans. Forty one percent of the nickel present in COB was extracted in a 3 L scale bioreactor (pH of 1.8 ± 0.05, temperature of 28 ± 2°C) maintained in anoxic environment. In contrast, only 11% of the nickel present in COB was extracted with continuous supply of air to the bioreactor keeping all the parameters unchanged. Kinetics study of anoxic microbial processing of COB revealed that the chemical reaction rate control model fits to the rate of nickel dissolution (R(2)=0.975).

Application of dolochar in the removal of cadmium and hexavalent chromium ions from aqueous solutions.

Dolochar, a waste material generated in sponge iron industry, is processed and put to test as an adsorbent for removal of Cd(II) and Cr(VI) ions from aqueous solutions. The dolochar samples were characterised to determine the different phases and their distribution by reflection microscopy. The analysis indicated that the sample consists of metallic iron, fused carbon, and Ca-Mg bearing phases (Ca-Mg-silicate-oxide) along with lots of voids and pores. The fixed carbon (FC) content of the material is 13.8% with a Langmuir surface area of 81.6m(2)/g and micropore area of 34.1m(2)/g. Batch adsorption experiments have been conducted to study the sorption behaviour of Cd(II) and Cr(VI) ions on dolochar as a function of particle size, contact time, adsorbent dosages, pH and temperature. It is observed that higher pH and temperature enhances sorption of Cd(II) ions. In contrast, the adsorption for Cr(VI) is found to be better in acidic pH in comparison to alkaline media. The equilibrium adsorption isotherm data are tested by applying both Langmuir and Freundlich isotherm models. It is observed that Langmuir isotherm model fitted better compared to the Freundlich model indicating monolayer adsorption. The thermodynamic parameters such as ΔG°, ΔH° and ΔS° indicate the effectiveness of dolochar to remove Cd(II) and Cr(VI) ions from aqueous solution. The kinetics of adsorption is found to better fit to pseudo second order reaction.

Fluoride adsorption studies on mixed-phase nano iron oxides prepared by surfactant mediation-precipitation technique.

Mixed nano iron oxides powder containing goethite (α-FeOOH), hematite (α-Fe(2)O(3)) and ferrihydrite (Fe(5)HO(8)·4H(2)O) was synthesized through surfactant mediation-precipitation route using cetyltrimethyl ammonium bromide (CTAB). The X-ray diffraction, FTIR, TEM, Mössbauer spectroscopy were employed to characterize the sample. These studies confirmed the nano powder contained 77% goethite, 9% hematite and 14% ferrihydrite. Fluoride adsorption onto the synthesized sample was investigated using batch adsorption method. The experimental parameters chosen for adsorption studies were: pH (3.0-10.0), temperature (35-55°C), concentrations of adsorbent (0.5-3.0 g/L), adsorbate (10-100 mg/L) and some anions. Adsorption of fluoride onto mixed iron oxide was initially very fast followed by a slow adsorption phase. By varying the initial pH in the range of 3.0-10.0, maximum adsorption was observed at a pH of 5.75. Presence of either SO(4)(2-) or Cl(-) adversely affected the adsorption of fluoride in the order of SO(4)(2-)>Cl(-). The FTIR studies of fluoride loaded adsorbent showed that partly the adsorption on the surface took place at surface hydroxyl sites. Mössbauer studies indicated that the overall absorption had gone down after fluoride adsorption that implies it has reduced the crystalline bond strength. The relative absorption area of ferrihydrite was marginally increased from 14 to 17%.

Physicochemical investigations of microemulsification of coconut oil and water using polyoxyethylene 2-cetyl ether (Brij 52) and isopropanol or ethanol.

The microemulsification of coconut oil/polyoxyethylene 2-cetyl ether/2-propanol or ethanol/water was investigated. The phase behaviors of the mixed system were examined. The shear viscosity at different temperatures was measured to derive activation parameters for the viscous flow. The diffusion coefficient of the microemulsions at different compositions was determined by the DLS method. The energetics of solubilization of water into oil + Brij + alkanol as well as of oil into water + Brij + alkanol forming w/o and o/w microemulsions, respectively, were calorimetrically determined.

The role of Wobe-Mugos in reducing acute sequele of radiation in head and neck cancers--a clinical phase-III randomized trial.

Oral enzymes act as a potent antiinflammatory, antiedematous agents thereby decreasing acute toxigenic effect of radiation and increasing compliance, quality of life of our patients. Fifty patients were randomized 25 allocated in enzyme and radiotherapy arm, 25 in radiotherapy alone. Pre RT and post RT biopsies were taken from both arms. In our study it was found that there was clinical, histopathological as well as statistical significant difference in both arms. The enzyme arm patients had mucostis of grade I in 76%, grade II in 12%, grade III in 8% while as 8% had grade I, 68% grade II, 24% had grade III in RT arm alone. In enzyme patients skin reactions of grade I in 72%, 20% had grade II, 8% had grade III. In control arm 12% had grade I, 76% had grade II, 8% had grade III skin reaction.