Characterization and optimization of lipase activity produced by Pseudomonas monteilli 2403-KY120354 isolated from ground beef

A total of 56 Gram negative bacterial isolates were recovered from twenty ground beef samples and were screened for their potentiality to produce lipase. Forty four bacterial isolates were recorded as positive producers for lipase on tween as carbon source in solid medium. Also, the highly producer isolates were screened for lipase activity in submerged culture using olive oil as carbon and the most active isolate was 2043 which gave an activity of 20.0 ± 0.29 U/ml. The bacterial isolate 2403 was identified phenotypically according to Bergey’s Manual and genotypically using 16S rRNA genes analysis as Pseudomonas monteilli. Effect of some different factors on lipase activity were studied and the maximum lipase activity was achieved at reaction medium of pH 6 and incubated at 40°C for 60 min. Also, addition of Ba2+ in the reaction medium enhanced the lipase activity, while the other tested metals reduced the enzyme activity.Key words: Food contamination, lipase activity, olive oil, cultural conditions, Pseudomonas

Comparative Molecular Transporter Properties of Cyclic Peptides Containing Tryptophan and Arginine Residues Formed through Disulfide Cyclization

We have previously reported cyclic cell-penetrating peptides [WR]5 and [WR]4 as molecular transporters. To optimize further the utility of our developed peptides for targeted therapy in cancer cells using the redox condition, we designed a new generation of peptides and evaluated their cytotoxicity as well as uptake behavior against different cancer cell lines. Thus, cyclic [C(WR)xC] and linear counterparts (C(WR)xC), where x = 4–5, were synthesized using Fmoc/tBu solid-phase peptide synthesis, purified, and characterized. The compounds did not show any significant cytotoxicity (at 25 µM) against ovarian (SK-OV-3), leukemia (CCRF-CEM), gastric adenocarcinoma (CRL-1739), breast carcinoma (MDA-MB-231), and normal kidney (LLCPK) cells after 24 and 72 h incubation. Both cyclic [C(WR)5C] and linear (C(WR)5C) demonstrated comparable molecular transporter properties versus [WR]5 in the delivery of a phosphopeptide (F′-GpYEEI) in CCRF-CEM cells. The uptake of F′-GpYEEI in the presence of 1,4-dithiothreitol (DTT) as the reducing agent was significantly improved in case of l(C(WR)5C), while it was not changed by [C(WR)5C]. Fluorescence microscopy also demonstrated a significant uptake of F′-GpYEEI in the presence of l(C(WR)5C). Cyclic [C(WR)5C] improved the uptake of the fluorescent-labeled anti-HIV drugs F′-d4T, F′-3TC, and F′-FTC by 3.0–4.9-fold. These data indicate that both [C(WR)5C] and linear (C(WR)5C) peptides can act as molecular transporters

Thymoquinone suppression of the human hepatocellular carcinoma cell growth involves inhibition of IL-8 expression, elevated levels of TRAIL receptors, oxidative stress and apoptosis

Hepatocellular carcinoma (HCC) is the fourth most common solid tumor worldwide. The chemokine interleukin-8 (IL-8) is overexpressed in HCC and is a potential target for therapy. Although the transcription factor NF-κB regulates IL-8 expression, and while thymoquinone (TQ; the most bioactive constituent of black seed oil) inhibits NF-κB activity, the precise mechanisms by which TQ regulates IL-8 and cancer cell growth remain to be clarified. Here, we report that TQ inhibited growth of HCC cells in a dose- and time-dependent manner, caused G2M cell cycle arrest, and stimulated apoptosis. Apoptosis was substantiated by activation of caspase-3 and -9, as well as cleavage of poly(ADP-ribose)polymerase. TQ treatments inhibited expression of NF-κB and suppressed IL-8 and its receptors. TQ treatments caused increased levels of reactive oxygen species (ROS) and mRNAs of oxidative stress-related genes, NQO1 and HO-1. Pretreatment of HepG2 cells with N-acetylcysteine, a scavenger of ROS, prevented TQ-induced cell death. TQ treatment stimulated mRNA expression of pro-apoptotic Bcl-xS and TRAIL death receptors, and inhibited expression of the anti-apoptotic gene Bcl-2. TQ enhanced TRAIL-induced death of HepG2 cells, in part by up-regulating TRAIL death receptors, inhibiting NF-κB and IL-8 and stimulating apoptosis. Altogether, these findings provide insights into the pleiotropic molecular mechanisms of TQ-dependent suppression of HCC cell growth and underscore potential of this compound as anti-HCC drug

Naringin attenuates the development of carrageenan-induced acute lung inflammation through inhibition of NF-κb, STAT3 and pro-inflammatory mediators and enhancement of IκBα and anti-inflammatory cytokines

Naringin has been reported to possess diverse pharmacological properties, including anti-arthritic and anti-inflammatory activities. The aim of the present study was to determine the potential anti-inflammatory effect of naringin in a mouse model of carrageenan-induced pleurisy. A single dose of naringin (40 and 80 mg/kg) was administered per oral (p.o.) 1 h before carrageenan (Cg) administration. Pro- and anti-inflammatory cytokines were analysed in pleural fluid. We also assessed the effects of naringin on the expression levels of iNOS, inducible cyclooxygenase isoform (COX-2), ICAM-1, MIP-2, PGE2, STAT3, TGF-β1, nuclear factor kappa B (NF-κB) and inhibitor of kappa B (IκBα) in lung tissue. The histological examinations revealed anti-inflammatory effect of naringin while Cg group deteriorated. Naringin downregulated Th1 and upregulated Th2 cytokines. Western blot analyses revealed increased protein expression of NF-κB, STAT3 and COX-2 and decreased IκBα in response to Cg treatment, which were reversed by the treatment with naringin. In the Cg group, mRNA expression levels of pro-inflammatory mediators upregulated and anti-inflammatory mediators downregulated. Naringin reversed these actions

Gene expression of IQGAPs and Ras families in an experimental mouse model for hepatocellular carcinoma: a mechanistic study of cancer progression

IQGAPs genes play critical role in either induction or suppression of cancer and its progression, however the relationship between Ras genes and these genes are still unclear. In this study, we tried to understand the mechanistic action of IQGAPs genes and its correlation with Ras genes in mouse hepatic cancer model. The genetic expressions of IQGAP1, IQGAP2, IQGAP3, Hras, Kras, Nras, Mras, Caspase3, and BAX were followed in both hepatocellular carcinoma and normal liver cells of Balbc mice. Genotoxic agent diethylnitrosamine (DEN)-induced hepatic cancer model was induced in male mice and recorded the occurrence of hepatocellular carcinoma by morphological and histological changes in the liver. It was observed that mRNA expressions of IQGAP1, Hras, Kras, Nras, Mras, Caspase3, and BAX genes were highly elevated in hepatocellular carcinoma cells when compared with normal liver cells, additionally their expressions increased by concentrating the dose of DEN. While, the expressions of IQGAP2 and IQGAP3 were significantly decreased in hepatocellular carcinoma cells when compared with normal liver cells, as well as their expressions decreased more with increasing the dose of DEN. It was concluded from this study that IQGAP1 has a strong signaling relationship with Ras genes in induction of cancer and it is considered as a key gene for induction or suppression of the hepatocellular carcinoma

Prevalence and Mycotoxigenic Potential of Fungi in Fish Feed Collected from Fish Farms in Egypt with a Particular Reference to Aflatoxins Contamination

  The current study is aimed to investigate the fungal contaminants in fish feed. Isolation of fungi was conducted on modified dichloran 18% glycerol agar (DG18) and potato dextrose agar (PDA). Feed samples were assayed for aflatoxins using HPLC. A total of 43 species belonging to 19 fungal genera recovered from 45 fish feed samples. Aspergillus and Penicillium were the most predominant genera with isolation frequency values indicated the retrieval capability of DG18 over PDA medium. For instance, Aspergillus spp. recorded 60%, 53.3% while Penicillium spp. were 33.3%, 17.8% on DG18 and PDA respectively via direct plating. 41.4% of the tested isolates were mycotoxin producers. Aflatoxins B1, B2, G1 and G2 were detected by 6 out of 10 screened Aspergillus isolates. Fumitremorgens, Gliotoxin, Ochratoxin A and B, and Zeralenone were also detected. The feed samples of high total count percentages (TC%) of A. flavus recorded the highest incidence of aflatoxins B2, G1 and G2 (2.3, 35.3 and 7.8 ng/g respectively). Meanwhile, the highest B1 concentration (3.7 ng/g) was recorded for the highest TC% interval studied (1:9 cfu/g). Thus, it is important to monitor the fungal load and mycotoxins in fish feed periodically using proper practical approaches

Prevalence and Mycotoxigenic Potential of Fungi in Fish Feed Collected from Fish Farms in Egypt with a Particular Reference to Aflatoxins Contamination

  The current study is aimed to investigate the fungal contaminants in fish feed. Isolation of fungi was conducted on modified dichloran 18% glycerol agar (DG18) and potato dextrose agar (PDA). Feed samples were assayed for aflatoxins using HPLC. A total of 43 species belonging to 19 fungal genera recovered from 45 fish feed samples. Aspergillus and Penicillium were the most predominant genera with isolation frequency values indicated the retrieval capability of DG18 over PDA medium. For instance, Aspergillus spp. recorded 60%, 53.3% while Penicillium spp. were 33.3%, 17.8% on DG18 and PDA respectively via direct plating. 41.4% of the tested isolates were mycotoxin producers. Aflatoxins B1, B2, G1 and G2 were detected by 6 out of 10 screened Aspergillus isolates. Fumitremorgens, Gliotoxin, Ochratoxin A and B, and Zeralenone were also detected. The feed samples of high total count percentages (TC%) of A. flavus recorded the highest incidence of aflatoxins B2, G1 and G2 (2.3, 35.3 and 7.8 ng/g respectively). Meanwhile, the highest B1 concentration (3.7 ng/g) was recorded for the highest TC% interval studied (1:9 cfu/g). Thus, it is important to monitor the fungal load and mycotoxins in fish feed periodically using proper practical approaches

Efficient Bioreduction of Sulfate from Industrial Wastewater Effluents Using Enterobacter cloacae emr69

This study aimed to isolate and characterize sulfur reducing bacteria from industrial wastewater and soil to remove sulfate. A total of 14 sulfate reducing bacterial (SRB) isolates were recovered from industrial wastewater and contaminated soil. Interestingly, bacterial isolate emr69 was selected as the highest sulfate reducer. Correspondingly, emr69 was characterized phenotypically and identified genotypically based on 16S rRNA gene sequencing as Enterobacter cloacae and deposited in Gen Bank database under accession number OR472728. The maximum sulfate reduction by E. cloacae emr69 against 2000 ppm (SO4-2) was 95% which obtained by adjusting the medium at pH 6 and growing the bacterium at 37°C under anaerobic conditions. The study suggests using of E. cloacae mr69 as a promising SRB for bioreduction of sulfate in industrial wastewater treatment

Optimization of lactic acid production by a novel strain, Enterococcus faecalis KY072975 isolated from infants stool in Egypt

Production of lactic acid using a novel strain of lactic acid bacteria isolated from infants stool was investigated in the present study. Out of ten isolates, a total of five bacterial isolates were found as positive in lactic acid production. The tested bacterial isolate W7 was observed as the potent strain in lactic acid production that exhibited a halo zone of 8 mm. The bacterial isolate W7 was identified phenotypically and genotypically as Enterococcus faecalis and was deposited in GenBank with accession number KY072975. The effect of different process parameters such as initial pH of the medium, incubation temperature, inoculum size and incubation time was also monitored to enhance lactic acid production and resulted in optimum lactic acid value of 0.72 mg/mL. The salted whey was the most applicable fermentation medium for production of lactic acid by Enterococcus faecalis KY072975 and recorded 2.07 ± 0.1 mg/mL

Investigation of the Pathological and Biochemical Characterizations in Naturally Infected Calves with Foot and Mouth Disease (FMD)

All cloven-hoofed animals are susceptible to foot-and-mouth disease (FMD) which is highly contagious viral illness. FMD is answerable for serious economic losses in Egypt. Despite the primary control approach being annual mass vaccination campaigns using polyvalent inactivated vaccinations, failure of vaccination has been according in several cases. The study was conducted on fifty native breed calves up to one year of age from both sexes, thirty calves suspected to be infected with FMD, and twenty clinically healthy calves were considered as controls. A total of fifty samples from organs (heart- epithelial tissue) were gathered from calves suspected to be FMD infected obtained from September 2021 to March 2022 in Sharkia and Menofeia governorates / Egypt. The current study was designed for the isolation of FMDV using BHK-21 cells. Molecular identification, through the extraction of Viral RNA, and RT-PCR were used to test samples for the FMDV virus. Diseased animals have changes in body temperature, respiration rate, and heart rate compared to controls. Moreover, murmur sounds were observed during auscultation of the heart. A hematological study revealed significant reductions in the RBCs count, hemoglobin concentration, and PCV% with leukopenia and lymphopenia in the diseased group. The serum cardiac troponin, lipase, non-esterified fatty acid, beta hydroxyl butyric acid, glucose, AST, ALT activities, and blood urea nitrogen were considerably enhanced in diseased animals. But serum insulin and amylase were significantly reduced in diseased calves. Histopathological examination of calves revealed extensive lymph histiocytic myocarditis and necrotic lesions in the pancreas, liver, and kidney. In conclusion, the early stages of FMD in calves is characterized by myocardial cell injury, elevation of blood cardiac troponin, and necrotic pancreatitis represented by atrophy of pancreatic glands and islets of Langerhans