Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus,hepatitis B virus,and human immunodeficiency virus 1

BACKGROUND Hepatitis C virus(HCV),hepatitis B virus(HBV),and human immunodeficiency virus 1(HIV-1)are the most epidemic blood-borne viruses,posing threats to human health and causing economic losses to nations for combating the infection transmission.The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive,but they are more accurate than sero-logical testing.AIM To develop a rapid,cost-effective,and accurate diagnostic multiplex polymerase chain reaction(PCR)assay for simultaneous detection of HCV,HBV,and HIV-1.METHODS The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electro-phoretic molecular weight inside each viral genome.Therefore,this diagnostic method will be appropriate for application in both conventional(combined with electrophoresis)and real-time PCR facilities.Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus.Then,Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay.RESULTS The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay.The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis.Compared to related published assays,the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays.CONCLUSION This study provides a simple,cost-effective,and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses;this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Effect of ion and electron beam irradiation on surface morphology and optical properties of PVA

Polyvinyl alcohol(PVA) is a well-known friendly polymer for paper-making, textiles, and a variety of coatings, biomedical applications such as artificial pancreas, synthetic vitreous body, wound dressing, artificial skin,and cardiovascular device. In this paper, ion/electron beam is employed to get insight into the irradiation effect on surface morphology and optical properties of PVA polymer. UV-Vis spectra are recorded to investigate the effect of induced defects on the optical band gap and the formed carbon clusters size. Scanning electron microscopy(SEM) is used to relate and investigate surface morphology and optical properties of the target polymer with different doses(15, 30 and 60 min). Also, PVA polymer is subjected to theoretical studies by using semi-empirical PM7 quantum chemical method.

Optimization of double chamber microbial fuel cell for domestic wastewater treatment and electricity production

Microbial fuel cells(M FCs)represent a new approach for treating w aste w ater along w ith electricity production.The present study addressed electricity production from domestic w astew ater using a mediator-less double chamber M FC.The electricity production w as monitored under different operational conditions for both summer and w inter samples.Optimization of the anodic and cathodic chambers resulted in a maximal current of 0.784 and 0.645 m A w ith the maximal pow er intensity of 209 and 117 m W/m2in pow er duration of 24 h for the summer and w inter samples,respectively.Scanning electron microscopy show ed that the bacterial biofilm formation on the anode w as denser for the summer sample than that w hen the w inter sample w as used,so w as the total bacterial count.Therefore,samples taken during summer w ere considered better in electricity production and w aste w ater treatment than those taken during w inter basically because of the high microbial load during the hot season.In parallel,there w as a decrease in both biological oxygen demand(BOD5)and chemical oxygen demand(COD)values which reached 71.8%and 72.85%,respectively at the end of the operation process for the summer sample,w hile there w as no evident decrease for the w inter sample.Optimizing the operating conditions not only increased the potential of using domestic w aste w ater in microbial fuel cells to produce electricity,but also improved the quality of the domestic w aste w ater.

Investigation of Dyed Film Based on Quinaldine Red Dyed Poly(Vinyl Alcohol) and Poly(Vinyl Butyral) for High Dose Dosimetry Applications

Dyed polymer films, prepared by a simple technique of casting aqueous solutions of poly(vinyl alcohol) PVA or poly(vinyl butyral) PVB containing quinaldine red (QR) on a horizontal glass plate, are useful as routine high-dose dosimeters. These flexible plastic film dosimeters are bleached when exposed to gamma rays. The response of these dosimeters depends on the concentration of QR and the polymer material. The radiation chemical yield (G-value) of both PVA and PVB dyed films was calculated and was found to increase with increasing dye concentration. The effect of relative humidity during irradiation as well as pre- and post-irradiation storage, on the response of the films is examined. These films are not affected by humidity change in the intermediate range of 10% - 50%.

Cytotoxic Effect of Chitosan Based Nanocomposite Synthesized by Radiation： In Vitro Liver and Breast Cancer Cell Line

A silver nanoparticle （AgNP） is likely to provide an attractive object for combining a variety of biochemical properties with great therapeutic potential by using radiation. The present study explores the ICs0 value of chitosan-poly （vinyl alcohol） hydrogel （Cs/PVA） and Ag-doped chitosan-poly （vinyl alcohol） （Cs/PVA/Ag） nanocomposite in view of their anticancer application. The aim was to develop （Cs/PVA） based hydrogel synthesized by gamma radiation which could behave both as a nanoreactor for Ag nanoparticle with promising anticancer applications. The （Cs/PVA/Ag） nanocomposite was confirmed by FTIR （Fourier transform infrared） spectroscopy, XRD （X-ray diffraction） and EDX （energy dispersive X-ray） analysis. The anti-cancer activity of the prepared nanocomposites was demonstrated in human liver cancer cell line （HEPG2） and breast cancer cell lines （MCF7）. It has significant effects against human liver cancer cell line HEPG2 compared to breast cancer cell line MCF7. Further quantitative analysis on the molecular and protein levels is still required to confirm the impact of chitosan on genotoxic effect before reaching a final conclusion and starting its biomedical application.

Influence of gamma-ray irradiation on optical and thermal degradation of poly (ethyl-methacrylate) (PEMA) polymer

Polymer based on (polyethyl-methacrylate) (PEMA) were exposed to different doses of gamma radiation up to 100 kGy and the physical properties have been studied. The effects of gamma irradiation on the optical spectrum of PEMA films have been investigated using spectrophotometric measurements of reflectance and transmittance in the wavelength range 200-1100 nm. The structure of the sample is analyzed by X-ray diffraction technique and is found to be amorphous and partially crystalline. TGA studies revealed that the thermal stability of polyethyl methacrylate, improved after irradiation doses up to 100 kGy. On other hand driving absorption coefficient α(ω), consequently the band tail width Ee and optical band gap estimated. This behavior is believed to be associated with the generation of excess of electronic localized states. Also, Optical constants such as refractive index (n), extinction coefficient (K) have been determined using Swanepole method. Optical dispersion parameters and the dispersion parameters, such as Eo (single–oscillator energy), Ed (dispersive energy) are discussed in terms of the single-oscillator Wemple-DiDomenico model.

Toluidine Blue O-Gelatin Gel Dosimeter for Radiation Processing

In this study the spectrophotometric response of the Toluidine Blue O (TBO)-gelatin gel dosimeter irradiated with gamma rays was characterized. Preparation of dyed-gelatin gel takes place in neutral medium;to give eventually gel dosimeter has a sharp absorbance peak at 635 nm that bleaches quantitatively upon irradiation and the colour change can be measured with UV-VIS spectrophotometer. The useful dose range was 1 - 150 Gy. The radiation chemical yield (G-value) of gel dosimeter was calculated and found to increase by increasing concentration of dye. Post-irradiation storage on the response of gel is discussed. The dose response function, radiation sensitivity, and dependences of the response on environmental factors were studied.

Heuristic techniques for maximum likelihood localization of radioactive sources via a sensor network

Maximum likelihood estimation(MLE)is an effective method for localizing radioactive sources in a given area.However,it requires an exhaustive search for parameter estimation,which is time-consuming.In this study,heuristic techniques were employed to search for radiation source parameters that provide the maximum likelihood by using a network of sensors.Hence,the time consumption of MLE would be effectively reduced.First,the radiation source was detected using the k-sigma method.Subsequently,the MLE was applied for parameter estimation using the readings and positions of the detectors that have detected the radiation source.A comparative study was performed in which the estimation accuracy and time consump-tion of the MLE were evaluated for traditional methods and heuristic techniques.The traditional MLE was performed via a grid search method using fixed and multiple resolutions.Additionally,four commonly used heuristic algorithms were applied:the firefly algorithm(FFA),particle swarm optimization(PSO),ant colony optimization(ACO),and artificial bee colony(ABC).The experiment was conducted using real data collected by the Low Scatter Irradiator facility at the Savannah River National Laboratory as part of the Intelligent Radiation Sensing System program.The comparative study showed that the estimation time was 3.27 s using fixed resolution MLE and 0.59 s using multi-resolution MLE.The time consumption for the heuristic-based MLE was 0.75,0.03,0.02,and 0.059 s for FFA,PSO,ACO,and ABC,respectively.The location estimation error was approximately 0.4 m using either the grid search-based MLE or the heuristic-based MLE.Hence,heuristic-based MLE can provide comparable estimation accuracy through a less time-consuming process than traditional MLE.

Flavone improves liver damage in nicotine-exposed rats via the Nrf2/HO-1 pathway

Objective:To assess the hepatoprotective effects of flavone on nicotine-induced liver damage.Methods:Thirty-six rats were allocated into six groups:the control group,the nicotine group,the flavone alone groups(10 and 25 mg/kg/body weight),and the nicotine groups treated with flavone(10 and 25 mg/kg/body weight).Liver function,oxidative stress,Nrf2 pathway(HO-1,Nrf2,and Keap-1),and inflammatory markers(IL-17,TNF-α,and NF-κB)were evaluated.Additionally,a histopathological examination of liver tissues was performed.Results:Nicotine increased liver damage,inflammation,and oxidative stress.However,flavone suppressed nicotine-induced liver enzymes,oxidative stress,and inflammation,as manifested by increased antioxidants and decreased malondialdehyde level,liver enzymatic activities,and inflammatory markers.Flavone(10 and 25 mg/kg/body weight)also reduced the level of Keap-1 and increased HO-1 and Nrf2 levels in the liver of nicotine-exposed rats.Conclusions:Flavone has hepatoprotective properties and may slow the progression of liver injury by reducing oxidative stress,liver enzymes,and inflammation possibly via the Nrf2 pathway.

Study Bacteriocin Production and Optimization Using New Isolates of <i>Lactobacillus</i>spp. Isolated from Some Dairy Products under Different Culture Conditions

Lactobacilli belong to the group of lactic acid bacteria (LAB), that have several distinguished abilities such as production of lactic acid, enzymes such as β-Galactosidase and natural antimicrobial substances called bacteriocins. Bacteriocin is a biopreservative agent potential of suppressing growth of some contaminant bacteria in food industry but its commercial availability is limited and costly. The study aimed to select isolates of Lactobacillus spp. potential for producing bacteriocins to suppress the growth of Escherichia coli ATCC 25922 and Bacillus subtilis NCIB3610, and to optimize the process of bacteriocin production. Results obtained in this study showed that L. acidophilus isolate CH1 was selected as the best candidate for bacteriocin among the four isolates that tested. The largest amounts of the bacteriocins were synthesized only in MRS medium was supplemented with K2HPO4 (1.0%), Tween 80 (1%), Beef extract (1%), glucose, cyctein and peptone extract (1%). The optimization of culture conditions for bacteriocin production areas showed that corn steep liquor medium was the best medium for all isolates against Bacillus subtilis while no effect was observed on Escherichia coli ATCC 25922 except when used MRS medium. The optimum conditions for bacteriocin production were pH 6.0, temperature 34?C with 4% Phenyl acetamide showing the greatest growth inhibition areas.

Expression levels of pro-inflammatory interleukin-8 and certain antimicrobial peptides in concurrent with bacterial conjunctivitis

AIM:To detect the quantitative expression levels of the pro-inflammatory interleukin-8(IL8),antimicrobial peptides human beta defense-2(HBD2),and human beta defense-3(HBD3)genes in bacterial conjunctivitis.METHODS:The human conjunctival epithelial cells were obtained using the impression cytology technique from healthy controls and patients.The genes expression levels were determined utilizing a reverse transcription quantitative polymerase chain reaction(RT-q PCR).The contribution of causative agent type,the number of isolates and severity of clinical features,in the increase of genes expression was also determined.RESULTS:The RT-q PCR showed that IL8,HBD2,and HBD3 expression increased in bacterial conjunctivitis as compared to healthy control(P<0.001).In gram-negative bacterial conjunctivitis,HBD2 was highly up-regulated(P<0.001)compared to other types of bacterial conjunctivitis.In mixed bacterial conjunctivitis,a direct correlation between HBD2 up-regulation and HBD3 up-regulation was observed(P<0.05).The severity of clinical features was related to the up-regulation of IL8 and HBD2(P<0.05).CONCLUSION:IL8,HBD2,and HBD3 are immuneeffectors in infectious conjunctivitis.HBD2 is active during different bacterial conjunctivitis but is more released with gram-negative bacteria compared to gram-positive bacteria.HBD3 is an obvious defender in different bacterial conjunctivitis.

Persea americana attenuates inflammatory response associated with hyperlipidemia in ovariectomized and irradiated rats by regulating MMP-3/TIMP-1 levels

Objective:To explore the effect of Persea americana supplementation on inflammation,oxidative stress,and lipid profiles in ovariectomized rats fed with a high-fat diet and exposed to radiation.Methods:The control group was sham operated,while groups 2-5 were ovariectomized and fed a high-fat diet.Groups 4 and 5 were exposed toγ-radiation(1 Gy/week for 5 weeks)after ovariectomy.Groups 3 and 5 were treated with 1 mL/250 g/day of Persea americana for one month.Serum levels of estrogen,alanine aminotransferase,aspartate aminotransferase,cholesterol,triglycerides and lipoproteins were measured.Additionally,hepatic oxidative stress,inflammatory and fibrogenic markers were evaluated.Results:Persea americana treatment reduced the oxidative stress markers as well as the levels of triglyceride,total cholesterol,and low-density lipoprotein cholesterol,which in turn lowered hepatic fat accumulation.Moreover,it suppressed hepatic inflammatory mediators(interleukin-6,tumor necrosis factor-α,and C-reactive protein)and downregulated pro-fibrogenic markers(transforming growth factor-βand tissue inhibitor of metalloproteinase-1).Conclusions:Persea americana provides protection against ovariectomy,and gamma radiation-mediated hepatic inflammation not only through its antioxidant,anti-inflammatory,lipid-lowering effect but also by modulating the fibrogenic markers.

Electrical,thermal and electrochemical properties of γ-ray-reduced graphene oxide

The properties of γ-ray-reduced graphene oxide samples(GRGOs)were compared with those of hydrazine hydrate-reduced graphene oxide(HRGO).Fourier transform infrared spectroscopy,X-ray diffractometry,Raman spectroscopy,Brunauer-Emmett-Teller surface area analysis,thermogravimetric analysis,electrometry,and cyclic voltammetry were carried out to verify the reduction process,structural changes,and defects of the samples,as well as to measure their thermal,electrical,and electrochemical properties.Irradiation with γ-rays distorted the structure of GRGOs and generated massive defects through the extensive formation of new smaller sp^(2)-hybridized domains compared with those of HRGO.The thermal stability of GRGOs was higher than that of HRGO,indicating the highly efficient removal of thermally-labile oxygen species by γ-rays.RRGO prepared at 80 kGy showed a pseudocapacitive behavior comparable with the electrical double-layer capacitance behavior of HRGO.Interestingly,the specific capacitance of GRGO was enhanced by nearly three times compared with that of HRGO.These results reflect the advantages of radiation reduction in energy storage applications.

Band Gap Determination from Diffuse Reflectance Measurements of Irradiated Lead Borate Glass System Doped with TiO₂by Using Diffuse Reflectance Technique

Ultraviolet-visible UV-Vis diffuse reflectance measurements of irradiated lithium tetra-borate glass have been acquired using. Evolution 600 Spectrophotometer equipped with praying Mantis Diffuse Reflectance Accessory DRA base upon the onset of the diffuse reflectance spectra of the powdered or bulk materials. Also the absorption edge and band gap energies of the prepared glass were determined. The optical energy gap is calculated and found to be (3.0 - 3.5) eV. Which is in close agreement to the one calculated for r = 1/2, i.e. the transition mechanism, is accordingly direct allowed transition. The density increases from 3.1 to 3.9 gm/cm3 for the undoped and doped glass.

Bioethanol Production from Rice Straw Enzymatically Saccharified by Fungal Isolates, Trichoderma viride F94 and Aspergillus terreus F98

Egypt faces a high population growth rate nowadays, which demands for an increase in agricultural production efficiency. Consequently, agricultural field residues will increase. Rice straw is one of the main agriculture residues in Egypt. So this study was performed on rice straw as a resource for production of bioethanol. Eight microbial isolates, five yeasts and three fungi were isolated from rice straw. Yeast isolates were selected for their ability to utilize different sugars and cellulose. Chipped and grinded rice straw was subjected to different pretreatment methods physically through steam treatment by autoclaving and different doses of gamma γ irradiation (50 and 70 Mrad). Autoclaved pretreated rice straw was further enzymatically treated throughout solid state fermentation process by different fungal isolates;F68, F94 and F98 producing maximum total reducing sugars of 12.62, 13.58, 17.00 g/L, respectively. Bioethanol production by separate microbial hydrolysis and fermentation (SHF) process of rice straw hydrolysate was performed by the two selected fungal isolates;Trichoderma viride F94 and Aspergillus terreus F98 and two yeast isolates (Y26 and Y39). The two yeast isolates have been identified by 18S, RNA as Candida tropicalis Y26 and Saccharomyces cerevisiae Y39. SHF processes by F94 and Y26 produced 45 gallon/ton rice straw while that of F98 and Y39 produced 50 gallon/ton rice straw.

Characterization and Antitumor Activity of Chitosan/Poly （Vinyl Alcohol） Blend Doped with Gold and Silver Nanoparticles in Treatment of Prostatic Cancer Model

The application of nanotechnology in various fields of science has earned a great concern over the past decades. The natural products and surface-modified polymers and metallic nanoparticles （NPs） have evolved as promising nanomaterials for targeted prostate cancer treatment. In the present study, Chitosan/poly （vinyl alcohol） （Cs/PVA） blend was synthesized by gamma radiation which could behave a nanoreactor for silver （Ag） and gold （Au） nanoparticle with promising anticancer applications. （Cs/PVA/Ag） and （Cs/PVA/Au） nanocomposites were confirmed by SEM （scanning electron microscope） and TEM （transmission electron microscope） analysis. The swelling properties have been investigated as a function of time and pH. The anti-cancer activity of the prepared nanocomposites was demonstrated in prostatic cancer cell line. It has a significant effect against prostatic cancer. However, metal nanoparticles have shown a good experimental success in the field of nanomedicine especially in cancer treatment, which has always been an area of high concern. The collaboration of biomedical research in the identification and characterization of biomedical strategies using the interesting metal nanocomposite will impact the future nanomedicine greatly.

Improvement of Hematopoietic and Immunologic Findings in Sublethal Gamma Irradiated Rats Treated with Bone Marrow Transplantation and Wheat Germ Oil

The objective of this study was to evaluate the efficacy of the wheat germ oil (WGO) and bone marrow transplantation (BMT) in boosting the immuno response and protecting from oxidative stress in irradiated rats. BM was given by intravenous injection to male rats, one hour post gamma irradiation at the dose level of 5 Gy. Rats were orally administrated with 54 mg/Kg body wt of wheat germ oil daily for 2 weeks before irradiation. After 14 days, results revealed that total body irradiation induced significant decreases in RBCs, WBCs and lymphocytes, as well as Glutathione (GSH) and zinc superoxide dismutase (Zn/SOD), splenocyte count, bone marrow lymphocyte count and viability. Tumor necrosis factor alpha (TNF-α) and interleukin 2 (IL-2) also recorded significant decrease while interleukin 6 (IL-6) and lipid peroxidation marker malondialdehde (MDA) in serum and spleen were conversely elevated. In irradiated animals receiving BMT and WGO, values of MDA in serum and tissue were significantly depressed as compared with the irradiated group, while lymphocytes, bone marrow viability percentage, splenocytes percentage, IL-2, IL-6 and GSH were significantly elevated. The curative action of WGO enforcing significant innate response could trigger and augment adaptive immune response by BMT, thus protecting immune system from radiation induced damage as well as oxidative stress.