In Vitro Effects of Natrum muriaticum in Kidney Cell Lines MDCK and LLC-PK1

Previous papers have indicated that homeopathic solutions modify the cellular and biochemical aspects of cells maintained in culture. In this study, the effects of Natrum muriaticum, a medicine used in the homeopathic clinic for the treatment of hypertension, were evaluated in kidney MDCK and LLC-PK1 cell lines. The following cellular parameters were analyzed: viability, morphology and expression of the (Na+-K+)-ATPase and the angiotensin II receptors AT1 and AT2. The cell lines were plated (5.0 x 104 cells/mL) in DMEM supplemented with 10% fetal calf serum (FCS). After 24 hours at 37°C, DMEM was re-fed with the addition of 10% (V/V) and 1% (V/V) of the following samples: Natrum muriaticum 30CH, water 30CH and non-dynamized sterile distilled water to do the MTT assay. The results obtained from these groups were compared to those obtained by incubation of the cells in culture medium free of these solutions (Control). Cell viability was assessed by a colorimetric MTT ELISA assay (490nm). The values from four independent experiments performed in quintuplicate were plotted and statistically analyzed by Sigma Plot v.11 (Jandel Scientific). The morphology of MDCK cells was evaluated by optical microscope after Giemsa’s staining. The expression of the (Na++K+)-ATPase and AT1/AT2 of LLC-PK1 cells was evaluated by Western Blot (WB) analysis. For this experimental set, 5.0x104 cells/mL were incubated in DMEM supplemented with 10% FCS and daily culture medium was replaced by a new one, containing: Natrum muriaticum 30CH and water 30CH. Additionally, cells were treated for 5, 10 and 15 days with 1% of specific solutions and the total protein was measured by the Lowry method, after cell lysis. The samples were analyzed by electrophoresis in SDS-PAGE (12% gel) and transferred to nitrocellulose membrane. This membrane was incubated with specific primary antibodies (anti-(Na++K+)-ATPase, anti-AT1 and AT2 or anti-anti-beta-actin). The detection was performed using ECL system and Hyperfilm. MTT assays showed a statistically significant reduction in cellular mitochondrial activity (p

In vitro behavior of Mycoplasmagallisepticum live-type nosode

As a step of a doctoral research project, in this study a live-type nosode was prepared from microorganism Mycoplasmagallisepticum strain R (ATCC 93-08/19610) according to Costa model and the rules by Brazilian Homeopathic Pharmacopoeia. Live nosode was tested in vitro to assess safety when used to immunize domestic fowl (Gallus gallus) against infection by this microorganism and to investigate its behavior under laboratory conditions. M. gallisepticum was not shown to grow in fluid (broth) and solid (plate) modified Frey medium with dilutions 11d, 12d, 20d and 30d. Inhibition halos about 2.0 mm were observed around paper disks impregnated with live-type nosode in microorganism-sown Petri dishes, whereas disks impregnated with conventional antibiotic oxytetracycline exhibited 8.0 mm inhibition halos. Protein assessment by Folin-Lowry method showed protein absence in dilutions 12d and 30d and neither microbial DNA traces were found in PCR assay in dilutions 12d, 20d and 30d.

Determination of Protein Oxidation in Aquaculture Feed

The aim of this research was to develop reliable, easy-to-perform and cheap method for measuring protein oxidation in complex samples such as aquaculture feed within various protein sources. For that purpose modified 2,4-dinitrophenylhydrazyne (DNPH)-based method for quantification of protein carbonyls was employed, while the method modification was consisted of using different solutions for the extraction, time of protein extraction and concentration of trichloracetic acid (TCA) for protein precipitation. It was found that extraction during night, higher TCA concentration and the use of 0.5 M KCl extraction solution resulted in the highest protein amount measured by Lowry method and 280 nm protein estimation. On the other hand, the lowest protein yield was obtained by using distillated water for the extraction. Furthermore, the lowest amount of protein carbonyls was in the case when extraction was performed with distilled water (DW), while the highest content of protein carbonyls were reached with 0.15 M KCl and 0.5 M KCl extraction solutions. It was observed that the amount of carbonyls compounds were increasing during storage under accelerated conditions, and in comparison to the original (unmodified) method, the modified method for measuring protein oxidation resulted in the higher amount of carbonyls during the all points of storage.

Influence of humic substances on the landfill leachate biodegradability with a focus on temporal seasonality

The high resilience to biological treatments from the landfill leachate is generally associated with the presence of humic substances (HS). The brown color characteristic of this effluent is also related to these substances. Landfill leachate with low biodegradability can make biological treatments unfeasible, which can drive up the cost for the treatment of large leachate volumes. In this context, this research aimed to characterize the leachate in different seasonal periods, and verify the influence of HS species on the biodegradability of the effluent to assist in the selection of adequate treatment techniques. The HS quantification was performed using the modified Lowry method and speciation through fractionation according to the molar masses of the HS species. The tropical regions can be the precursor for the rapid stabilization of biodegradable organic matter. The warmer climate contributed to a reduced BOD/COD ratio (0.03) and the predominance of compounds of lower mass (e.g.: fluvic acids). The tests showed an HS concentration of 26.9% of the total COD in the raw leachate in the rainy season, which increased to 37.3% in the dry season. Approximately 70% of HS species refer to fulvic acids, a fraction identified as having the highest biologic treatment resilience.

Differences in the standardization of medicinal products based on extracts of chondroitin sulfate

Objective: a comparative analysis of 6 different chondroprotectors for injection, containing chondroitin sulfate (CS) based on natural extracts.Material and methods. Five samples were studied for each CS extract. The preparations were compared on the basis of profiles of trace element composition, sulfur content, chromatographic analysis and test for total protein. In each sample, the concentration of 72 elements was determined, then averaging was performed and the variances of the content of each element were calculated. To assess the content of the protein fraction, a modified Lowry method with bicincholic acid was used. Chromatographic profiles of the studied CS extracts were measured to estimate the molecular weight distribution.Results. The studied samples differ significantly from each other in the total and individual content of sulfur, toxic and conditionally toxic microelements. According to the analysis of sulfur and trace elements, a cluster of more standardized CS extracts was identified. It was shown that the assessments of the pharmaceutical quality of the studied extracts made by the elemental profile, correspond to the assessments of the quality by the degree of proteins elimination and by the molecular weight characteristics of CS extracts. In particular, the highest total content of toxic elements was found for CS-6 (14.87±1.81 μg/l) and CS-2 (9.20±1.12 μg/l), and the lowest – for CS-4 (1.46±0.23 μg/l), CS-3 (1.92±0.33 μg/l) and CS-1 (2.98±0.25 μg/l). The highest content of protein impurities was also found in CS-6 (9.62 mg/ml) and CS-2 (6.64 mg/ml), and the lowest – in CS-1 (2.87 mg/ml). At the same time, the highest amount of sulfur was found CS-1 (6400 mg/kg) and much less – for CS-2 (370 mg/kg) and CS-6 (100 mg/kg). Significant amounts of the high-molecular fraction of CS (1–40 kDa) were found only in CS-1, and only trace amounts of high-molecular CS forms were present in CS-2 and CS-6.Conclusion. The highest content of cholesterol and sulfur and, at the same time, the lowest content of toxic microelements and proteins were distinguished by the extract obtained from the trachea of a bovine.

Ketahanan Gubal Jati Hutan Rakyat Diawetkan dengan Senyawa Boron Menggunakan Metode Tekan Lowry terhadap Serangan Rayap Tanah dan Kayu Kering

Nowadays, most of teak wood that available on the market was mainly from comunity forest. Those teak woods were commonly harvested in the early age. The young teak wood from community forests possesses lower proportion of heartwood than sapwood. Those condition was generally had an affect on the wood durability. Although the wood durability was estimated to be low, but the teak wood from community forest was intensively used for furniture and house construction. Therefore, study on the sapwood durability of young teak wood from community forest is necessary. In the present study, the durability of sapwood treated by boron compound (boric acid and borax) as wood preservatives was investigated. The boron concentrations were 5, 7 and 10%. The vacuum process were used to impreg the preservative into the wood specimens called as Lowry method. The efectivity of preservation method were investigated, namely absorption, retention, penetration, efficacy of subterranean and dry-wood termites. The absorption, retention, and penetration of boron compound were 69.10–96.41 kg/m3, 4.53–5.31 kg/m3, and 3.04–3.16 mm, respectively. Absorption, retention, and penetration of preservatives showed an increasing value by increasing the preservatives concentrations, with the highest values were obtained for 10% boron concentration. Graveyard test was used to evaluate the efficacy of boron in teak sapwood to subterranean termites. During 2 months observation, mass loss and degree of wood damage were 0.42-1.37% and 6.31-18.72%. Furthermore, the efficacy of boron was also conducted for dry-wood termites. The mass loss, degree of wood damage, and dry-wood termites mortality after 28 days observation were 1.46-1.67%, 29.45-32.38%, and 87.33-95.33%, respectively. The durability of boron treatened teak sapwood against subterranean and dry-wood termites was increased. The increasing of teak sapwood durability was characterized by reduction of mass loss and degree of wood damage compared to untreated teak sapwood (control).

Polyphenol rich mung bean (Vigna radiata) yogurt for obesity prevention

This study was aimed to obtain yogurt formula rich in antioxidant phenolic and soluble protein of mung beans. Mung beans were washed, soaked, and then drained. With a ratio of 1:8, mung-beans and water, blended, and then filtered to produce mung beans juice. Mung beans yogurt (Mungbe-Yo) were made with the proportion of mung bean juice and skim milk were 100:0; 95:5; 90:10, and 85:15, before added with sugar as much as 10%, pasteurized in a 70°C temperature for 20 mins. After that, each Mungbe-Yo formula was cooled to 45°C, then added lactic acid bacteria (LAB) as a starter, as much as 2% and 4%, followed by incubation at room temperature for 24 hrs. Sensory tests included color, taste, flavor, and viscosity were performed by fifty-five panelists on a 5-point score basis. Antioxidant phenolic content was measured using the Folin-Ciocalteu method, while soluble protein was using the Lowry method. The highest score and most likable color, flavor, viscosity, and taste by panelists were formula P3B2, it was not different significantly with P3B1, P2B1, P2B2, and P1B2 (p>0.05). The proportion of mung bean juice-skim milk and LAB significantly affected the total phenolic and soluble protein (P<0.05), meanwhile its interaction was also statistically significant (P<0.05). Using effectivity index, the best Mungbe-Yo formula according to its total phenol and soluble protein content, as well as sensory was P3B1 formula, which was 85% mung bean juice, 15% skim milk and 2% LAB, containing phenolics and soluble protein were 525.958±48.9 mg GAE/L and 43.179±1.87%, respectively. Mungbe-Yo is suitable for individuals with obesity.

Penambahan putih telur sebagai peningkat protein pada tempe kedelai

Efforts to increase protein in Tempe can provide positive values for native Indonesian food. The amount of remaining egg white in the bread industry can be used in making soybean Tempe. Egg white which is rich in albumin has never been used in the manufacture of soybean tempeh by the producer. This research aims to determine the effect of adding raw and steamed egg whites on the protein content of soybean Tempe. This study used a Completely Randomized Design (CRD) with 3 replications. The concentrations of egg whites used in the research included 0%, 2%, 4%, 6%, 8%, and 10%. The research instrument used was the observation sheet for protein content using the Lowry method. The research data were dissolved protein content. The data analysis technique used Two Way ANOVA. The results of the research showed that there was no effect of adding raw and steamed egg whites on soybean Tempe protein content (P > 0.05). However, the addition of raw egg whites can increase the protein content of soybean Tempe, compared to steamed egg whites. The conclusion of this research is the addition of raw and steamed egg whites does not have a significant effect on the protein content of soybean Tempe. The right type of egg white flour to increase the protein content of soybean Tempe is raw egg white flour with a protein content of 15.0 per 100 grams.

Antimicrobial Activity of Non-bond Colicin on Candida albicans Biofilm

Two hundred fifty mid-stream urine specimens were collected from Baqubah Teaching Hospital and Al-Batool Teaching Hospital from patients with urinary tract infections (UTI). Of these investigated urine specimens, 66 (26.4%) specimens showed positive growth culture of Gram-negative bacteria. From these, Escherichia coli was the most prevalent bacteria of the examined culture (41, 62.12%). Additionally, the cup assay was used to determine colicin producers while the most efficient colicin producers were estimated by the formation of larger inhibition zone. Approximately half of the investigated E. coli isolates (20, 49 %) was colicin producers. Colicins was extracted after induction by mitomycin-C showed a concentration of 3020 μg/ml, as estimated utilizing the Lowry method, while its activity was 80 U/ml. Our study results showed that colicin had significant antibiofilm activity (P≤ 0.05) against Candida albicans and the effect seemed to be concentration dependent. . However, the values of biofilm inhibition varied depending on the different tested isolates. The biofilm of isolate 5 showed the most significant inhibition (P≤ 0.05) by colicin with a value of 46%, while isolate 3 was less affected with an inhibition rate of 19% at the concentration of 2500 μl/ml.

Freeze-thaw system for thermostable β-Galactosidase isolation from Gedong Songo Geobacillus sp. isolate

The effective isolation of intracellular enzymes from thermophilic bacteria is challenging because of their sturdy membrane. On the other hand, the low-cost and nontoxic method is essential for industrial food enzymes. The freeze-thaw cycles using acetone-dry ice as a frozen system was studied for efficient isolation of thermostable b-galactosidase from Geobacillus sp. dYTae-14. This enzyme has been known for application in the dairy industry to reduce the lactose content. In this study, the freeze-thaw method was performed with cycle variations 3, 5, and 7 cycles. Acetone-dry ice (-78°C) is used as a frozen system and boiling water for thawing. The b-galactosidase activity was assayed using ortho-Nitrophenyl-β-galactoside (ONPG) as substrate and protein content determined with the Lowry method. The results show that the most effective freeze-thaw is five cycles. The enzyme’s highest specific activity is 3610.13 units/mg proteins at 40-60 % ammonium sulfate saturation, with a purity value of 2.52.