Copper oxide nanoparticles (CuO-NPs) as a key player in the production of oil-based paint against biofilm and other activities.

Copper oxide nanoparticles are among the metal nanoparticles gaining popularity in many biotechnological fields, particularly in marine environments. Their antimicrobial and antibiofilm activities make them appealing to many researchers. Among the various methods of producing nanoparticles, biosynthesis is crucial. Thus, a large number of reports have been made about the microbiological manufacture of these nanoparticles by bacteria. Nevertheless, bio-production by means of the cell-free supernatant of marine bacteria is still in its primary phase. This is landmark research to look at how bacteria make a lot (14 g/L) of copper oxide nanoparticles (CuO-NPs) via the cell-free supernatant of Bacillus siamensis HS, their characterization, and their environmental and medical approaches. The biosynthesized nanoparticles were characterized using a UV-visible spectrum range that provides two maximum absorption peaks, one obtained at 400 nm and the other around 550-600 nm. Diffraction of X-rays (XRD) clarifies that the size of the NPs obtained was estimated to be 18 nm using Debye-Scherrer's equation. Scanning electron microscope-energy dispersive X-ray spectroscopy (SEM-EDX) displays 91.93 % copper oxide purity. The Transmission Electron Microscope (TEM) image proves that the particles have a spherical form and an average diameter of 6.54-8.60 nm. At the environmental level, nanoparticles incorporated into oil-based paint can be used as antibiofilm tools to diminish the biofilm formed on the submerged surface in the marine environment. In disease management, NPs can be used as a wound healing agent to reduce the wound gap size as well as an anti-tumour agent to control liver cancer cells (hepatoma cells (HepG2)).

Production, characterization and biomedical potential of biosurfactants produced by haloalkaliphilic archaea from Wadi El-Natrun, Egypt.

Extreme halophilic archaea that can live in high saline environments can offer potential applications in different biotechnological fields. This study delves into the fascinating field of halophilic archaea and their ability to produce biosurfactants. Some strains of haloarchaea were isolated from Wadi El-Natrun and were screened for biosurfactants production in a standard basal medium using emulsification index assay. Two strains were chosen as the potential strains for surface tension reduction. They were identified as Natrialba sp. BG1 and N3. The biosurfactants production was optimized and the produced emulsifiers were partially purified and identified using FTIR and NMR. Sequential statistical optimization, Plackett-Burman (PB) and Box-Behnken Designs (BBD) were carried out using 5 factors: oil, NaCl, casamino acids, pH, and inoculum size. The most significant factors were used for the next Response Surface Methodology experiment. The final optimal conditions for biosurfactants production were the inoculum size 2% pH 11 and NaCl 250 g/L, for Natrialba sp. BG1 and inoculum size 2.2%, pH 10 and NaCl 100 g/L for Natrialba sp. N3. The produced biosurfactants were tested for wound healing and the results indicated that Natrialba sp. BG1 biosurfactants is more efficient than Natrialba sp. N3 biosurfactants. Biosurfactants extracts were tested for their cytotoxic effects on normal cell line as well as on different cancer cells using MTT assay. The findings demonstrated that varying concentrations of the biosurfactants (31.25, 62.5, 125, 250, 500 and 1000 µg/mL) exhibited cytotoxic effects on the cell lines being tested. Additionally, the outcomes unveiled the presence of anti-inflammatory and antioxidant properties for both biosurfactants. Consequently, they could potentially serve as natural, safe, and efficient novel agents for combating cancer, promoting wound healing, and providing anti-inflammatory and antioxidant benefits.

Some novel bioactivities of Virgibacillus halodenitrificans carotenoids, isolated from Wadi El-Natrun lakes.

Carotenoids come in second among the most frequent natural pigments and are utilized in medications, nutraceuticals, cosmetics, food pigments, and feed supplements. Based on recent complementary work, Virgibacillus was announced for the first time as a member of Wadi El-Natrun salt and soda lakes microbiota, identified as Virgibacillus halodenitrificans, and named V. halodenitrificans DASH; hence, this work aimed to investigate several in vitro medicinal bioactivities of V. halodenitrificans DASH carotenoids. The carotenoid methanolic extract showed antioxidant activity based on diphenylpicrylhydrazyl (DPPH) scavenging capacity with a half-maximal concentration (IC50) of 1.6 mg/mL as well as nitric oxide (NO) scavenging action expressed by an IC50 of 46.4 µg/mL. The extract showed considerable inhibitory activity for alpha-amylase (α-amylase) and alpha-glucosidase (α-glucosidase) enzymes (IC50 of 100 and 173.4 µg/mL, respectively). Moreover, the extract displayed selective anticancer activity against Caco-2 (IC50 = 138.96 µg/mL) and HepG-2 cell lines (IC50 = 31.25 µg/mL), representing colorectal adenocarcinoma and hepatoblastoma. Likewise, the extract showed 98.9 % clearance for human hepatitis C virus (HCV) using reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), HCV-NS5B polymerase activity inhibition (IC50 = 27.4 µg/mL), and selective inhibitory activity against human coronavirus (HCoV 229E) using the plaque reduction assay (IC50 = 53.5 µg/mL). As far as we can tell, the anticancer, antiviral, and antidiabetic attributes of Virgibacillus carotenoids are, de novo, reported in this work which accordingly invokes further exploration of the other medicinal, biotechnological, and industrial applications of Virgibacillus and haloalkaliphilic bacteria carotenoids.

Identification of potent anti-Candida metabolites produced by the soft coral associated Streptomyces sp. HC14 using chemoinformatics.

Candida albicans is the most common pathogen responsible for both spontaneous and recurrent candidiasis. The available treatment of Candida infections has several adverse effects, and the development of new drugs is critical. The current study looked at the synthesis of anti-Candida metabolites by Streptomyces sp. HC14 recovered from a soft coral. Using the Plackett Burman design, the medium composition was formulated to maximize production. Using GC-MS, the compounds have been identified, and a cheminformatics approach has been used to identify the potential source of activity. The compounds that showed high potential for activity were identified as pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(phenylmethyl)-3 and di-n-octyl based on their docking score against the cytochrome monooxygenase (CYP51) enzyme in Candida albicans. As a result of their discovery, fewer molecules need to be chemically synthesized, and fermentation optimization maximizes their synthesis, providing a strong foundation for the development of novel anti-Candida albicans agents.

Production, purification, and characterization of cold-active lipase from the psychrotroph Pseudomonas sp. A6.

Cold-active lipases are presently employed extensively in the detergent, chemical intermediate, fine chemical, food, and pharmaceutical industries. Seven cold-adaptive bacteria were isolated from the Mediterranean Sea near Alexandria, Egypt, and tested for their ability to produce cold-active lipase, with the highest activity at 10 °C. The most potent isolate was Pseudomonas sp. A6. To determine the most important variables, the bacterium was exposed to a necessary medium component and environmental factor screening using a single factor-at-a-time approach, followed by a multifactorial Plackett-Burman design strategy. After purification and characterization, the optimal activity levels for the cold-active lipase were figured out. Inoculation of Pseudomonas A6 under near optimum conditions using medium consisting of (g/L) peptone 7.14; soybean oil 7.5% (v/v); K2HPO4, 0.4; MgSO4, 0.1; glucose 2; pH 8; and temperature 10 °C led to a maximum lipase activity anticipated to be 23.36 U/mL. Purified lipase showed the best activity and thermal stability at a pH of 8 and a temperature of 10 °C. The Pseudomonas A6 lipase tolerated the monovalent ions, while greater valence ions did not.

Biosynthesis, characterization and optimization of TiO2 nanoparticles by novel marine halophilic Halomonas sp. RAM2: application of natural dye-sensitized solar cells.

BACKGROUND: Metal oxide nanoparticles (NPs) are becoming valuable due to their novel applications. The green synthesis of TiO2 NPs is more popular as a flexible and eco-friendly method compared to traditional chemical synthesis methods. TiO2 NPs are the most commonly used semiconductor in dye-sensitized solar cells (DSSCs). RESULTS: The biogenic TiO2 NPs were produced extracellularly by the marine halophilic bacterium Halomonas sp. RAM2. Response surface methodology (RSM) was used to optimize the biosynthesis process, resulting in a starting TiO2 concentration of 0.031 M and a pH of 5 for 92 min (⁓15 nm). TiO2 NPs were well-characterized after the calcination process at different temperatures of 500, 600, 700 and 800 °C. Anatase TiO2 NPs (calcined at 500 °C) with a smaller surface area and a wider bandgap were nominated for use in natural dye-sensitized solar cells (NDSSCs). The natural dye used as a photosensitizer is a mixture of three carotenoids extracted from the marine bacterium Kocuria sp. RAM1. NDSSCs were evaluated under standard illumination. After optimization of the counter electrode, NDSSCBio(10) (10 layers) demonstrated the highest photoelectric conversion efficiency (η) of 0.44%, which was almost as good as NDSSCP25 (0.55%). CONCLUSION: The obtained results confirmed the successful green synthesis of TiO2 NPs and suggested a novel use in combination with bacterial carotenoids in DSSC fabrication, which represents an initial step for further efficiency enhancement studies.

Bioprocess development for biosurfactant production by Natrialba sp. M6 with effective direct virucidal and anti-replicative potential against HCV and HSV.

Halophilic archaea is considered an promising natural source of many important metabolites. This study focused on one of the surface-active biomolecules named biosurfactants produced by haloarchaeon Natrialba sp. M6. The production trend was optimized and the product was partially purified and identified using GC-Mass spectrometry. Sequential optimization approaches, Plackett-Burman (PB) and Box-Behnken Designs (BBD) were applied to maximize the biosurfactants production from M6 strain by using 14 factors; pH, NaCl, agitation and glycerol; the most significant factors that influenced the biosurfactant production were used for Response Surface Methodology (RSM). The final optimal production conditions were agitation (150 rpm), glycerol (3%), NaCl (20.8%), pH (12) and cultivation temperature (37°C). GC-Mass spectrometry for the recovered extract revealed the presence of a diverse group of bipolar nature, hydrophobic hydrocarbon chain and charged function group. The majority of these compounds are fatty acids. Based on results of GC-MS, compositional analysis content and Zetasizer, it was proposed that the extracted biosurfactant produced by haloarchaeon Natrialba sp. M6 could be a cationic lipoprotein. The antiviral activity of such biosurfactant was investigated against hepatitis C (HCV) and herpes simplex (HSV1) viruses at its maximum safe doses (20 µg/mL and 8 µg/mL, respectively). Its mode of antiviral action was declared to be primarily via deactivating viral envelopes thus preventing viral entry. Moreover, this biosurfactant inhibited RNA polymerase- and DNA polymerase-mediated viral replication at IC50 of 2.28 and 4.39 µg/mL, respectively also. Molecular docking studies showed that surfactin resided well and was bound to the specified motif with low and accepted binding energies (ΔG = - 5.629, - 6.997 kcal/mol) respectively. Therefore, such biosurfactant could be presented as a natural safe and effective novel antiviral agent.

Carotegenic Virgibacillus halodenitrificans from Wadi El-Natrun Salt Lakes: Isolation, Optimization, Characterization and Biological Activities of Carotenoids.

Carotenoids, as phytonutrient pigments, are signified by their unique beneficial features that serve human health and the surrounding ecosystem. Haloalkaliphiles from soda lakes produce different natural bioactive molecules; however, their ability to produce carotenoids has been limited. Therefore, this study focused on the screening and isolation of carotenoid-producing haloalkaliphilic microbes. Out of 10 isolates, a powerful carotigenic bacterium was isolated, characterized phenotypically and identified on the molecular level as Virgibacillus halodenitrificans. By employing statistical approaches like Plackett-Burman design and central composite design, the influence of significant nutritional variables on carotenoids production was screened and optimized. Predictive modeling manifested that carotenoid yield was 36.42 mg/mL, a 2.12-fold enhancement compared to the basal conditions through inoculating 1.8% of bacterial biomass on optimized medium containing yeast extract (2 g/mL), peptone (10 g/mL) and NaCl (233.6 g/mL). The carotenoids content was confirmed by UV-Vis spectrum which detected a characteristic unique peak with left and right shoulders at 461 nm, 490 and 522 nm. However, FTIR and Raman spectroscopy showed the presence of several functional groups. Meanwhile, LC-MS confirmed that the examined carotenoids were composed of ß-carotene, lutein and ß-Apo-8'-carotenal mixture. As a bioactive agent, the carotenoids of V. halodenitrificans DASH showed characteristic antagonistic potency against a wide spectrum of Gram-positive and Gram-negative pathogens. Interestingly, a potent antifungal capacity was observed against Candida albicans, reflecting promising mycocidal efficacy against COVID-19 white fungal post-infections. Furthermore, carotenoids (20 µg/mL) inhibited the biofilm formation of P. aeruginosa and S. aureus by 54.01 ± 3.97% and 80.082 ± 0.895%, respectively. Our results proposed that haloalkaliphiles of Wadi El-Natrun lakes are promising sources of carotenoids that exhibited efficiency as safe, biocompatible and natural bioactive agents for environmental, medical and industrial applications.

Optimization and multiple in vitro activity potentials of carotenoids from marine Kocuria sp. RAM1.

Marine pigmented bacteria are a promising natural source of carotenoids. Kocuria sp. RAM1 was isolated from the Red Sea Bohadschia graeffei collected from Marsa Alam, Egypt, and used for carotenoids production. The extracted carotenoids were purified by thin-layer chromatography (TLC). The characteristic UV absorbance of the three purified fractions gave us an inkling of what the purified pigments were. The chemical structures were confirmed by nuclear magnetic resonance spectroscopy (NMR) and LC-ESI-QTOF-MS/MS. The three different red pigments were identified as two C50-carotenoids, namely bisanhydrobacterioruberin and trisanhydrobacterioruberin, in addition to 3,4,3',4'-Tetrahydrospirilloxanthin (C42-carotenoids). Kocuria sp. RAM1 carotenoids were investigated for multiple activities, including antimicrobial, anti-inflammatory, antioxidant, anti-HSV-1, anticancer, antidiabetic and wound healing. These new observations suggest that Kocuria sp. RAM1 carotenoids can be used as a distinctive natural pigment with potent properties.

Toxicity monitoring of solvents, hydrocarbons, and heavy metals using statistically optimized model of luminous Vibrio sp. 6HFE.

BACKGROUND: The utilization of bioluminescent bacteria in environmental monitoring of water contaminates considers being a vital and powerful approach. This study aimed to isolate, optimize, and apply luminescent bacteria for toxicity monitoring of various toxicants in wastewater. RESULTS: On the basis of light intensity, strain Vibrio sp. 6HFE was initially selected, physiologically/morphologically characterized, and identified using the 16SrDNA gene. The luminescence production was further optimized by employing statistical approaches (Plackett-Burman design and central composite design). The maximum bioluminescence intensity recorded 1.53 × 106 CPS using optimized medium containing (g/L), yeast extract (0.2g), CaCl2 (4.0), MgSO4 (0.1), and K2HPO4 (0.1) by 2.3-fold increase within 1h. The harnessing of Vibrio sp. 6HFE as a bioluminescent reporter for toxicity of organic solvents was examined using a bioluminescence inhibition assay. According to IC50 results, the toxicity order of such pollutants was chloroform > isoamyl > acetic acid > formamide > ethyl acetate > acetonitrile > DMSO > acetone > methanol. However, among eight heavy metals tested, the bioluminescence was most sensitive to Ag+ and Hg+ and least sensitive to Co2+ and Ni2+. Additionally, the bioluminescence was inhibited by benzene, catechol, phenol, and penta-chlorophenol at 443.1, 500, 535.1, and 537.4 ppm. CONCLUSION: Vibrio sp. 6HFE succeeded in pollution detection at four different environmental and wastewater samples revealing its efficiency in ecotoxicity monitoring.

Biosynthesis of Silver Nanoparticles by Aspergillus terreus: Characterization, Optimization, and Biological Activities.

In this study, mycelial filtrate of Aspergillus terreus BA6 was used to reduce AgNO3 to form silver nanoparticles (AgNPs). The effect of seven independent variables on the diameter of AgNPs was studied by applying design of experiments (DOE). At optimal conditions, the diameter of AgNPs was reduced by approximately 26.7% compared to the basal culture condition and AgNO3 concentration was found to be the most significant factor affecting the diameter of AgNPs. A. terreus nano-Ag was characterized using UV-visible spectroscopy, transmission electron microscopy, energy dispersive X-ray (EDX), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and Zeta potential. The maximum UV absorption was obtained at 420 nm and the microscopic results showed particles with narrow size distribution ranging from 7 to 23 nm. XRD pattern of AgNPs revealed four diffraction peaks of metallic silver and the EDX spectrum showed a strong signal attributed to Ag nano-crystals. AgNPs mycofabricated by A. terreus showed potent minimum inhibitory concentration (MIC) and broad minimum bactericidal/fungicidal concentration (MBC/MFC) against 12 reference microorganisms. The MIC and MBC/MFC values of AgNPs were 0.312 to 1.25 µg/ml and 0.625 to 10 µg/ml, respectively. Nevertheless, AgNPs did not demonstrate any antagonistic activity against Coxsackie B virus. The in vitro cytotoxicity of the mycosynthesized AgNPs showed significant antitumor activity against adenocarcinoma epithelial cells from human breast cancer (Mcf-7) cell line with an inhibitory concentration (IC50) of 87.5 µg/ml.

Large scale production of superparamagnetic iron oxide nanoparticles by the haloarchaeon Halobiforma sp. N1 and their potential in localized hyperthermia cancer therapy.

Magnetic iron oxide nanoparticles are among metal nanoparticles that attract huge attention in many biotechnological fields especially in the biomedical area. Their extensive capabilities and easy separation methodology drive them to be an interesting point to many researchers. Biosynthesis is of a major importance among different methods of nanoparticles production. Microbial synthesis of these nanoparticles by bacteria and yeasts have been reported on a wide scale. However, biosynthesis using halophilic archaea is still in an early stage. This study reveals the first contribution of the haloarchaeon Halobiforma sp. N1 to the nanobiotechnology field. It reports a rapid and economical one-step method of fabricating functionalized superparamagnetic iron oxide nanoparticles and their feasibility for hyperthermia treatment for cancer therapy. Herein, we have focused on optimizing the quantity of these fascinating nanoparticles, obtaining a very high yield of 15 g l-1 with high dispersion in water solution. Their unique characteristics enable them to participate in medical applications. They are nearly spherical in shape with a high degree of homogenity and uniformity with average diameter of 25 ± 9 nm. Also, the magnetic properties and elemental structure of the formed nanoparticles tend to be superparamagnetic like behavior with saturation magnetization of 62 emu g-1 and purity of 98.38% of iron oxide, respectively. The specific absorption rate (SAR) was measured and the particles induced significant heating power at lower frequencies which is a promising result to be applied for in vitro/in vivo hyperthermia studies in the near future.

In vitro dual (anticancer and antiviral) activity of the carotenoids produced by haloalkaliphilic archaeon Natrialba sp. M6.

Halophilic archaea are a promising natural source of carotenoids. However, little information is available about the biological impacts of these archaeal metabolites. Here, carotenoids of Natrialba sp. M6, which was isolated from Wadi El-Natrun, were produced, purified and identified by Raman spectroscopy, GC-mass spectrometry, and Fourier transform infrared spectroscopy, LC-mass spectrometry and Nuclear magnetic resonance spectroscopy. The C50 carotenoid bacterioruberin was found to be the predominant compound. Because cancer and viral hepatitis are serious diseases, the anticancer, anti-HCV and anti-HBV potentials of these extracted carotenoids (pigments) were examined for the first time. In vitro results indicated that the caspase-mediated apoptotic anticancer effect of this pigment and its inhibitory efficacy against matrix metalloprotease 9 were significantly higher than those of 5-fluorouracil. Furthermore, the extracted pigment exhibited significantly stronger activity for eliminating HCV and HBV in infected human blood mononuclear cells than currently used drugs. This antiviral activity may be attributed to its inhibitory potential against HCV RNA and HBV DNA polymerases, which thereby suppresses HCV and HBV replication, as indicated by a high viral clearance % in the treated cells. These novel findings suggest that the C50 carotenoid of Natrialba sp. M6 can be used as an alternative source of natural metabolites that confer potent anticancer and antiviral activities.

Phylogenetic diversity and antimicrobial activity of marine bacteria associated with the soft coral Sarcophyton glaucum.

Coral reefs are the most biodiverse and biologically productive of all marine ecosystems. Corals harbor diverse and abundant prokaryotic groups. However, little is known about the diversity of coral-associated microorganisms. We used molecular techniques to identify and compare the culturable bacterial assemblages associated with the soft coral Sarcophyton glaucum from the Red sea. Different media were utilized for microbial isolation, and the phylogeny of the culturable bacteria associated with the coral was analyzed based on 16S rDNA sequencing. The coral associated bacteria were found to be representatives within the Gammaproteobacteria, Actinobacteria, and Firmicutes. Antimicrobial activities of twenty bacterial isolates were tested against four pathogenic bacteria (Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa, Vibrio fluvialis) and three fungi (Penicillium sp., Aspergillus niger, Candida albicans). A relatively high proportion of bacterial strains displayed distinct antibacterial and antifungal activities, suggesting that soft coral-associated microorganisms may aid their host in protection against marine pathogens. Members of genera Bacillus and Pseudomonas had the highest proportion of antimicrobial activity which supported the hypothesis that they might play a protective role in the coral hosts.

Hepatitis C virus-mediated angiogenesis: molecular mechanisms and therapeutic strategies.

Angiogenesis is an essential process for organ growth and repair. Thus, an imbalance in this process can lead to several diseases including malignancy. Angiogenesis is a critical step in vascular remodeling, tissue damage and wound healing besides being required for invasive tumor growth and metastasis. Because angiogenesis sets an important point in the control of tumor progression, its inhibition is considered a valuable therapeutic approach for tumor treatment. Chronic liver disease including hepatitis C virus (HCV) infection is one of the main cause for the development of hepatic angiogenesis and thereby plays a critical role in the modulation of hepatic angiogenesis that finally leads to hepatocellular carcinoma progression and invasion. Thus, understanding of the molecular mechanisms of HCV-mediated hepatic angiogenesis will help design a therapeutic protocol for the intervention of HCV-mediated angiogenesis and subsequently its outcome. In this review, we will focus on the molecular mechanisms of HCV-mediated hepatic angiogenesis and the related signaling pathways that can be target for current and under development therapeutic approaches.

Hepatitis C virus-related hepatocellular carcinoma: An insight into molecular mechanisms and therapeutic strategies.

Hepatitis C virus (HCV) infects more than 170 million people worldwide, and thereby becomes a series global health challenge. Chronic infection with HCV is considered one of the major causes of end-stage liver disease including cirrhosis and hepatocellular carcinoma. Although the multiple functions of the HCV proteins and their impacts on the modulation of the intracellular signaling transduction processes, the drive of carcinogenesis during the infection with HCV, is thought to result from the interactions of viral proteins with host cell proteins. Thus, the induction of mutator phenotype, in liver, by the expression of HCV proteins provides a key mechanism for the development of HCV-associated hepatocellular carcinoma (HCC). HCC is considered one of the most common malignancies worldwide with increasing incidence during the past decades. In many countries, the trend of HCC is attributed to several liver diseases including HCV infection. However, the development of HCC is very complicated and results mainly from the imbalance between tumor suppressor genes and oncogenes, as well as from the alteration of cellular factors leading to a genomic instability. Besides the poor prognosis of HCC patients, this type of tumor is quite resistance to the available therapies. Thus, understanding the molecular mechanisms, which are implicated in the development of HCC during the course of HCV infection, may help to design a general therapeutic protocol for the treatment and/or the prevention of this malignancy. This review summarizes the current knowledge of the molecular mechanisms, which are involved in the development of HCV-associated HCC and the possible therapeutic strategies.

Hepatitis C virus-host interactions: Etiopathogenesis and therapeutic strategies.

Hepatitis C virus (HCV) is a significant health problem facing the world. This virus infects more than 170 million people worldwide and is considered the major cause of both acute and chronic hepatitis. Persons become infected mainly through parenteral exposure to infected material by blood transfusions or injections with nonsterile needles. Although the sexual behavior is considered as a high risk factor for HCV infection, the transmission of HCV infection through sexual means, is less frequently. Currently, the available treatment for patients with chronic HCV infection is interferon based therapies alone or in combination with ribavirin and protease inhibitors. Although a sustained virological response of patients to the applied therapy, a great portion of patients did not show any response. HCV infection is mostly associated with progressive liver diseases including fibrosis, cirrhosis and hepatocellular carcinoma. Although the focus of many patients and clinicians is sometimes limited to that problem, the natural history of HCV infection (HCV) is also associated with the development of several extrahepatic manifestations including dermatologic, rheumatologic, neurologic, and nephrologic complications, diabetes, arterial hypertension, autoantibodies and cryglobulins. Despite the notion that HCV-mediated extrahepatic manifestations are credible, the mechanism of their modulation is not fully described in detail. Therefore, the understanding of the molecular mechanisms of HCV-induced alteration of intracellular signal transduction pathways, during the course of HCV infection, may offer novel therapeutic targets for HCV-associated both hepatic and extrahepatic manifestations. This review will elaborate the etiopathogenesis of HCV-host interactions and summarize the current knowledge of HCV-associated diseases and their possible therapeutic strategies.

Hepatitis C virus core protein triggers hepatic angiogenesis by a mechanism including multiple pathways.

Chronic hepatitis C virus (HCV) infection is associated with the production of serum cytokines, including transforming growth factor (TGF)-beta2. Despite the occurrence of hepatic angiogenesis in liver conditions, the role of HCV proteins in this context is currently unknown. We demonstrated that the development of hepatic neoangiogenesis in patients infected with HCV is associated with the expression of TGF-beta2 and vascular endothelial growth factor (VEGF) and with activation of endothelial cells, as evidenced by CD34 expression. The analysis of liver biopsies of HCV-positive and HCV-negative patients using immunostaining showed significant elevation of TGF-beta2, VEGF, and CD34 expression in patients who were HCV-positive. Using an HCV established culture system, we confirmed further the production of both TGF-beta2 and VEGF proteins, in the hepatoma cell lines HepG2 and Huh7 by transfection with full-length HCV RNA (JFH1) or by the regulated expression of core. In addition, regulated expression of core protein in HepG2 or Huh7 cells was found to induce expression and activation of the transcription factor E2F1 and apoptosis signal-regulating kinase 1 (ASK1), activation of c-jun-N-terminal kinase (JNK) and p38, and extracellular-regulated kinase (ERK), and transcription factors activator protein 1 (AP-1), activating transcription factor 2 (ATF-2), cyclic adenosine monophosphate response element binding (CREB), E2F1, hypoxia inducing factor 1 alpha (HIF-1alpha), and specificity protein 1. Furthermore, data obtained from inhibitor experiments revealed the importance of E2F1 and ASK1 in the modulation of core-induced activation of JNK and p38 pathways and suggested an essential role for JNK, p38, and ERK pathways in the regulation of core-induced production of TGF-beta2 and VEGF proteins. Thus, our data provide insight into the molecular mechanisms whereby core protein mediates the development of hepatic angiogenesis in patients with chronic HCV infection.

Induction of high-molecular-weight (HMW) tumor necrosis factor(TNF) alpha by hepatitis C virus (HCV) non-structural protein 3 (NS3) in liver cells is AP-1 and NF-kappaB-dependent activation.

Chronic infection of hepatitis C virus (HCV)-infected patients is associated with the production of serum and interhepatic inflammatory cytokines including tumor necrosis factor alpha (TNF-alpha). In this study, we delineated part of the mechanism whereby HCV induces the synthesis of TNF-alpha in human liver cell lines HepG2 and Huh7. HepG2 transiently transfected with the full-length HCV cDNA expressed high-molecular-weight (HMW) TNF-alpha mRNAs, which were absent in the control cells. In addition tightly regulated expression of HCV NS3 in both HepG2 and Huh7 was found to induce the expression of HMW mRNAs and subsequently the production of biologically active TNF-alpha. Interestingly, the expression of NS3 protein in HepG2-NS3 or in Huh7-NS3 resulted in the activation of kinase (IKK-alpha) of NF-alphaB inhibitor (IalphaB) and in the enhancement of the DNA-binding activity of the nuclear transcription factor NF-kappaB. The inhibition of the transcription of TNF-alpha mRNAs and subsequently TNF-alpha production following the treatment of HepG2-NS3 or Huh7-NS3 transfectants with the inhibitor of NF-kappaB, Bay 11-7082, suggesting the importance of NF-kappaB for the regulation of NS3-mediated TNF-alpha expression in HepG2 and HeLa cells. Interestingly, data obtained from luciferase assays, in liver and in non-liver cells showed the contribution of NS3 protein in the regulation of TNF-alpha promoter through the activation of AP-1 and NF-kappaB. Our data indicate that the intrahepatic TNF-alpha production induced by HCV is transcriptionally up-regulated by HCV NS3. Therefore, HCV NS3 may have a potential role in the induction of intrahepatic inflammatory processes that occur during acute and chronic hepatitis C.

Biodiversity of moderately halophilic bacteria in hypersaline habitats in Egypt.

Screening bacteria from different saline environments in Alexandria. Egypt, lead to the isolation of 76 Gram-negative and 14 Gram-positive moderately halophilic bacteria. The isolates were characterized taxonomically for a total of 155 features. These results were analyzed by numerical techniques using simple matching coefficient (SSM) and the clustering was achieved by the unweighed pair-group method of association (UPGMA). At 75% similarity level the Gram-negative bacteria were clustered in 7 phena in addition to one single isolate, whereas 4 phena represented the Gram-positive. Based on phenotypic characteristics, it is suggested that the Gram-negative bacteria belong to the genera Pseudoalteromonas, Flavobacterium, Chromohalobacter, Halomonas and Salegentibacter, in addition to the non-identified single isolate. The Gram-positive bacteria are proposed to belong to the genera Halobacillus, Salinicoccus, Staphylococcus and Tetragenococcus. This study provides the first publication on the biodiversity of moderately halophilic bacteria in saline environments in Alexandria, Egypt.