Characterization of Enzyme Produced from Pseudomonas Putida for BTX (Benzene, Toluene & Xylene) Treatment in Petrochemical Industry Wastewater System

One of the big challenges in petrochemical industries is waste management. Currently, huge money was spending on the disposal of the waste. Industries are trying hard to find an alternative method to reduce the cost and improve the effectiveness of current waste management including treatment efficiency. Most of petrochemical wastes are containing benzene, toluene and xylene (BTX) which are very harmful to environment and living organisms. Common method used to separate the BTX from the waste are by using liquid-liquid and stripping process. One of the alternative to treat BTX is biological treatment method that used the natural capability of microorganisms to degrade to less harmful product is been applied. Some of examples are Pseudomonas Putida. (P. putida), Rhizobium, and Agrobacterium. P. putida is selected in this study for the biological treatment of BTX in petrochemical wastewater because it can produce an enzyme that has the capability of breakdown the aromatic hydrocarbon to carbon dioxide (CO2) and water (H2O). The main objective of this study is to produce and extract the enzymes produce, characterised the enzymes. This study also to investigate the effect of different concentration on the treatment as well as the growth of the bacteria. The enzyme is purified using salt precipitation and analysed using SDS_PAGE technique. UV-Vis is used to study the growth of the bacteria in the culture stock by measuring its optical density. The concentration of BTX was varied to determine the effect of the concentration on the percentage removal and the growth of P. putida. Enzymes detected or purified in this study was benzene reductase. Other expected enzymes were not able to be purified or analysed. It was found that in this experimental study, the removal of benzene is at 74% to 80%. The removal of toluene is at 62% to 75%. The removal of xylene is at 23% to 42%. Increasing the concentration of contaminants will reduce the removal capabilities

Septicaemia models using Streptococcus pneumoniae and Listeria monocytogenes: understanding the role of complement properdin

Streptococcus pneumoniae and Listeria monocytogenes, pathogens which can cause severe infectious disease in human, were used to infect properdin-deficient and wildtype mice. The aim was to deduce a role for properdin, positive regulator of the alternative pathway of complement activation, by comparing and contrasting the immune response of the two genotypes in vivo. We show that properdin-deficient and wildtype mice mounted antipneumococcal serotype-specific IgM antibodies, which were protective. Properdin-deficient mice, however, had increased survival in the model of streptococcal pneumonia and sepsis. Low activity of the classical pathway of complement and modulation of FcγR2b expression appear to be pathogenically involved. In listeriosis, however, properdin-deficient mice had reduced survival and a dendritic cell population that was impaired in maturation and activity. In vitro analyses of splenocytes and bone marrow-derived myeloid cells support the view that the opposing outcomes of properdin-deficient and wildtype mice in these two infection models is likely to be due to a skewing of macrophage activity to an M2 phenotype in the properdin-deficient mice. The phenotypes observed thus appear to reflect the extent to which M2- or M1-polarised macrophages are involved in the immune responses to S. pneumoniae and L. monocytogenes. We conclude that properdin controls the strength of immune responses by affecting humoral as well as cellular phenotypes during acute bacterial infection and ensuing inflammation

The apoptotic effects of Brucea javanica fruit extract against HT29 cells associated with p53 upregulation and inhibition of NF-κB translocation

Elham Bagheri,1 Fatemeh Hajiaghaalipour,2 Shaik Nyamathulla,1 Nur’Ain Salehen3 1Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; 2Institute of Biological Science, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; 3Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia Background: Brucea javanica (L.) Merr. is a plant from the genus Brucea, which is used in local traditional medicine to treat various diseases. Recent studies revealed an impressive anticancer efficiency of B. javanica extract in different types of cancer cells.Purpose: In this study, we have investigated the cytotoxic effects of the B. javanica hexane, ethanolic extracts against colon cancer cells. HT29 colon cells were selected as an in vitro cancer model to evaluate the anticancer activity of B. javanica ethanolic extract (BJEE) and the possible mechanisms of action that induced apoptosis.Methods: 3-(4,5-dimethylthiazol-2-yl)-2, 5,-diphenyltetrazolium bromide (MTT), lactate dehydrogenase, acridine orange/propidium iodide, and annexin-V-fluorescein isothiocyanate assays were performed to determine the antiproliferative and apoptosis validation of BJEE on cancer cells. Measurement of reactive oxygen species (ROS) production, caspase activities, nucleus factor-κB activity, and gene expression experiments was done to investigate the potential mechanisms of action in the apoptotic process.Results: The results obtained from this study illustrated the significant antiproliferative effect of BJEE on colorectal cancer cells, with a concentration value that inhibits 50% of the cell growth of 25±3.1 µg/mL after 72 h of treatment. MTT assay demonstrated that the BJEE is selectively toxic to cancer cells, and BJEE induced cell apoptosis via activation of caspase-8 along with modulation of apoptosis-related proteins such as Fas, CD40, tumor necrosis factor-related apoptosis-inducing ligands, and tumor necrosis factor receptors, which confirmed the contribution of extrinsic pathway. Meanwhile, increased ROS production in treated cells subsequently activated caspase-9 production, which triggered the intrinsic pathways. In addition, overexpression of cytochrome-c, Bax, and Bad proteins along with suppression of Bcl-2 illustrated that mitochondrial-dependent pathway also contributed to BJEE-induced cell death. Consistent with the findings from this study, BJEE-induced cancer cell death proceeds via extrinsic and intrinsic mitochondrial-dependent and -independent events.Conclusion: From the evidence obtained from this study, it is concluded that the BJEE is a promising natural extract to combat colorectal cancer cells (HT29 cells) via induction of apoptosis through activation of extrinsic and intrinsic pathways. Keywords: Brucea javanica, apoptosis, cancer, HT29, mitochondrial pathway, apoptosis protein arra

Assessment of wound healing potential of copper (II) Bis [N'-((5-chloro-1H-indol-3-YL) methylene) nicotinohydrazide] on experimentally-induced excision wounds in rats

Copper (II) Schiff base derivatives are frequently used for the treatment of many disorders. The aim of current study was to investigate the effects of the Schiff base derivatives on rats of wound healing enclosure and histology of granulation tissues in rats. Wound healing activity of Copper (II) Bis [N'-((5-chloro-1H-indol-3-YL) methylene) nicotinohydrazide] which is shortly called CI-indole-nicotinic, was perused by specifing the percentage of wound closure after tropical application of the compound. Four groups of animals were treated respectively with 0.2mL of gum acacia, 0.2mL of Intrasite gel, 0.2mL of CI-indole-nicotinic (50 mg/mL) and 0.2mL of CI-indole-nicotinic (100 mg/mL) for 10 days to assess the gross rate of wound enclosure, histology and endogenous enzymes parameters. The results indicated the affective capacity of CI-indole-nicotinic inj tissue regenetration and healing the injured skin. Grossing on day 5 and day 10 declared that the compound significantly accelerated the rate of wound healing. Histology of granulation tissues indicated remarkably the angiogenesis augmentation and increased fibroblasts and collagen formation as well as reduction in flammatory cells balance in CI-indole-nicotinic-treated groups compared to control group. Moreover, the compund represented dose-dependent levels of superoxide dismutase (SOD) and slight lipid peroxidation enzyme - malondialdedhyde (MDA) inhibition in granulation tissue. The acute toxity examination displayed no nephorotoxic or hepatotoxic side effects. In conclusion, wound healing capability of CI-indole-nicotinic compund could possibly attribute to enhance collagen deposition in granulation tissue and antioxidant activity of the compound

Gastroprophylactic Effects of p-Cymene in Ethanol-Induced Gastric Ulcer in Rats

The prevalence of gastric ulcers has increased in recent years, mainly because of non-steroidal anti-inflammatory drug utilization. Therefore, the current study investigates the gastroprotective effect of p-Cymene on absolute ethanol-induced acute gastric mucosal hemorrhagic lesions in rats. Thirty Sprague Dawley rats were randomly separated into five groups: normal control, ulcer control, reference, and two experimental groups. The normal and ulcer control groups were orally fed with 0.5% carboxymethylcellulose (CMC). The reference group was fed orally with 20 mg/kg omeprazole. The experimental groups were fed with 30 mg/kg and 60 mg/kg p-Cymene, respectively. After one hour, the normal group was fed with 0.5% CMC, and groups 2–5 were given absolute alcohol. After another hour all rats were sacrificed. The ulcer control group showed severe superficial hemorrhagic gastric mucosal lesions with decreased gastric mucus secretion and pH of gastric content. p-Cymene significantly reduced ethanol-induced gastric lesions, as evidenced by increased mucus and pH of gastric content, decreased ulcer area, reduced or absence of edema, and leucocyte infiltration of the subcutaneous layer. In gastric mucosal homogenate, p-Cymene displayed a significant increase in superoxide dismutase (SOD), catalase (CAT) activities, prostaglandin E2 (PGE2), and significantly reduced the malondialdehyde (MDA) level. In addition, p-Cymene increased the intensity of periodic acid–Schiff (PAS) stain of the gastric epithelium, and produced up-regulation of the HSP 70 protein and down-regulation of the Bax protein of the stomach epithelium, as well as a reduction in the levels of tumor necrotic factor-alpha and interleukin-6, while the level of interleukin-10 was increased. p-Cymene decreased the level of TNF-a and IL-6, and increased the level of IL-10. Acute toxicity with a higher dose of 500 mg/kg p-Cymene did not manifest any toxicological signs in rats and could enhance defensive mechanisms against gastric mucosal lesions. p-Cymene showed gastroprotective effects that could be attributed to its antioxidant nature, or its ability to increase mucus secretion, increase endogenous enzymes (SOD, CAT, PGE2), reduce MDA level, up-regulate HSP 70 protein, down-regulate Bax protein, and modulate inflammatory cytokines