A green chemical analysis of iron (III) in water sample using a smart phone.

In this study, a smartphone was used as the photometer for determination of iron (III) by forming a complex with thiocyanate. After color formation at concentrations of, image capture with mobile phone, signal analysis of each sample was performed by the application and converted to the absorption number. The calibration curve was completely linear in the range of 10 to 80 mg L-1 used and the linear coefficient was better than 0.9833. The limits of detection (LOD) and quantification (LOQ) were 0.1 and 0.3 mg L-1, respectively. Finally, this method was successfully used to measure iron in real samples.•A smartphone was used for the determination of iron (III), showcasing its potential in color compound analysis.•The method demonstrated superior performance in terms of calibration curve range and measurement speed compared to traditional atomic absorption devices.•The technique was successfully applied in the measurement of iron in real samples, indicating its practical applicability.

Extraction and purification of oleuropein from olive leaves.

In this study, effect of some solvents (ethanol, acetonitrile and water) for the extraction of phenolic compounds was investigated. Solvent volume, temperature and duration of the extraction process have a positive correlation with the amounts of phenolic compounds and antioxidant activity. The amounts of phenolic compounds, especially oleuropein in the leaf extract is higher than the fruit extract, and the concentration of oleuropein content in leaves and fruits extract was 4900 and 1800 mg/L, respectively. The oleuropein extracted from olive leaves was purified with liquid-liquid extraction to above 98%, and its concentration in leaves was 9.8 w/w/% in dried leaves.

Optimization of Ziziphora clinopodioides L. essential oil nanoencapsulation in chitosan nanocomplex by response surface methodology.

Nano-encapsulation of essential oils, a specific area of interest, can help overcome challenges associated with their commercial use. This study aimed to evaluate the effect of different concentrations of chitosan, Ziziphora clinopodioides L. essential oil (ZcEO), and Sodium-Tri Polyphosphate (TPP), both individually and in interaction, on several properties of EO-loaded chitosan nanoparticles. These properties include particle size (PS), zeta potential (ZP), and encapsulation efficiency (EE) using a two-stage emulsion-ionic gelation approach. The optimization of the parameters was done by response surface methodology using Box-Behnken design. The chemical composition of ZcEO was analyzed as well. The primary compounds in ZcEO were found to be pulegone (29.24 %), 1,3-dimethyl-2-(2-methylpropylidene) imidazolidine (9.05 %), piperitenone (6.65 %), thymol (5.38 %), and carvacrol (5.27 %). The PS ranged from 117.33 to 4934.1 nm, the ZP varied from -1.1 to -30.83 mV, and the EE spanned from 31.74 to 87.04 %. The results showed that an increase in the initial EO content led to a decrease in PS and ZP, but an increase in EE. Moreover, increasing the TPP concentration resulted in an enhancement in PS, ZP, and EE, whereas increasing the Chs concentration led to a slight increase in PS, ZP, and EE. Furthermore, the results of this study proved the interaction effect of different parameters on the responses investigated. Under optimized conditions, the optimal concentrations of chitosan, ZcEO and TPP were attained at 6.768, 6.078, and 7.595 mg/mL respectively. This resulted in a PS of 117.331 nm, a ZP of -20.949 mV, and an EE of 75.385 %. In conclusion, the results suggest that adjusting the concentrations of Chs, EO, and TPP is an effective approach to controlling the properties of NPs and optimizing their performance.

In Vivo Toxicity of Industrial Biocide Containing 2,2-Dibromo-3-nitrilopropionamide in Adult and Zebrafish Larvae.

2,2-Dibromo-3-nitrilopropionamide (DBNPA) is a widely used biocide with potential environmental implications due to its toxicity. This study aimed to investigate the in vivo toxicity of DBNPA in zebrafish (Danio rerio), a model organism in environmental toxicology. Both adult and larval zebrafish were exposed to varying concentrations of DBNPA, and significant morphological changes and mortality rates were observed. The study found that even relatively low concentrations of DBNPA can have detrimental effects on zebrafish embryonic development, and high concentrations resulted in rapid mortality in adult zebrafish and larvae. The LC50 values calculated from this study were 9.3 ppm for adults and 9.1 ppm for larvae, indicating the high toxicity of DBNPA to these organisms. These findings underscore the potential environmental impact of DBNPA and highlight the need for further research into its effects on aquatic ecosystems.

Development and evaluation of cell membrane-based biomimetic nanoparticles loaded by Clostridium perfringens epsilon toxin: a novel vaccine delivery platform for Clostridial-associated diseases.

As Clostridium perfringens (C. perfringens) epsilon toxin (ETX) ranks as the third most potent clostridial toxin after botulinum and tetanus toxins, vaccination is necessary for creatures that can be affected by it to be safe from the effects of this toxin. Nowadays, nanostructures are good choices for carriers for biological environments. We aimed to synthesize biomimetic biodegradable nanodevices to enhance the efficiency of the ETX vaccine. For this purpose, poly(lactic-co-glycolic acid) (PLGA) copolymer loaded with purified epsilon protoxin (proETX) to create nanoparticles called nanotoxins (NTs) and then coated by RBC membrane-derived vesicles (RVs) to form epsilon nanotoxoids (RV-NTs). The resulting RV-NTs shaped smooth spherical surfaces with double-layer core/shell structure with an average particle size of 105.9 ± 35.1 nm and encapsulation efficiency of 97.5% ± 0.13%. Compared with NTs, the RV-NTs were more stable for 15 consecutive days. In addition, although both structures showed a long-term cumulative release, the release rates from RV-NTs were slower than NTs during 144 hours. According to the results of cell viability, ETX loading in PLGA and entrapment in the RBC membrane decreased the toxicity of the toxin. The presence of PLGA enhances the uptake of proETX, and the synthesized structures showed no significant lesion after injection. These results demonstrate that NTs and RV-NTs could serve as an effective vaccine platform to deliver ETX for future in vivo assays.

Catalytic degradation of propetamphos and azamethiphos using silver ion.

In this study, the degradation of two organophosphate pesticides, namely, propetamphos and azamethiphos, in the presence of Ag+ at different mole ratios was investigated. Moreover, the kinetic and degradation pathways of both chemicals in the range of 0-60 min were explored. Gas chromatography equipped with a thermionic specific detector was used to investigate the pesticide degradation kinetics and mechanism. The results show that the degradation rate of both pesticides follows first-order kinetic. The first-order rate constant and the half-life of reaction were in the range of 0.002-0.143 min-1, 187-2.1 min, and 0.005-0.164 min-1 and 60-1.8 min, for propetamphos and azamethiphos, respectively, at ambient temperature (25 ºC). Because group containing sulfur atom is a better leaving group than group containing nitrogen, the rate of degradation of azamethiphos is higher than propetamphos. In a higher mole ratio of Ag+ to pesticides, the degradation rate was increased, and it is possible to predict the rate of degradation of pesticides according to the chemical composition of leaving group.

Magnetic Beads of Zero Valent Iron Doped Polyethersolfun Developed for Removal of Arsenic from Apatite-Soil Treated Water.

The drop immerses calcium chloride aqueous solution was utilized to prepare the zero valent iron-doped polyethersulfone beads (PES/ZVI) for the efficient removal of arsenic from apatite-soil treated waters. The proposed beads can assist in promoting uptake efficiency by hindering ZVI agglomeration due to a high porosity and different active sites. The PES/ZVI beads were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and vibrating sample magnetism (VSM). The main objective of this study was to investigate the function of new PES/ZVI beads with an increased removal efficiency for the remediation of arsenic ions from the apatite-soil treated waters. A maximum adsorption removal of 82.39% was achieved when the experiment was performed with 80 mg of adsorbent for a contact time of 180 min. Based on the results, a removal efficiency >90% was obtained after 300 min of shaking time with an arsenic concentration of 20 mg·L-1. The experimental process was fitted with the Langmuir model due to the high R2 (0.99) value compared to the Freundlich model (0.91) with an adsorption capacity of 41.32 mg·g-1. The adsorption process speed was limited by pseudo-second-order (R2 = 0.999) and the adsorption mechanism nature was endothermic and physical.

Determination of furfurals in baby food samples after extraction by a novel functionalized magnetic porous carbon.

Herein, a novel polypyrrole-polyaniline functionalized magnetic porous carbon (MPC@PPy-PANI) composite material was fabricated and utilized for the separation/extraction of furfurals from baby food and dry milk samples. In this way, magnetite@silica nanoparticles were first synthesized, and then a magnetic metal-organic framework (MMIL-101(Fe)) was prepared. After that, the MMIL-101(Fe) was pyrolyzed in a neutral atmosphere to obtain MPC. Ultimately, the MPC was functionalized with a co-polymer of aniline-pyrrole via oxidation polymerization. The synthesis of MPC@PPy-PANI was confirmed with FT-IR spectroscopy, SEM, TEM, VSM, and XRD techniques. Furfural and hydroxymethyl furfural were selected as the model analytes, which were separated/quantified on an HPLC-UV instrument. The LODs, LOQs, and linear dynamic ranges (LDRs) were in the range of 0.3-0.7 µg kg-1, 1.0-2.5 µg kg-1, and 1.0-600 µg kg-1, respectively. Repeatability of the method was studied as an RSD parameter, and was located in the range of 5.5-6.8% (within-day, n = 5) and 8.2-9.4% (between-day, n = 3 days). The applicability of the proposed method was established by analyzing several baby food and dry milk samples. The relative recovery (RR%) and repeatability were located in the range of 86-111% and 3.3-10.1%, respectively, showing excellent accuracy and precision of the method.

Simian virus 40 DNA in immunocompetent children with respiratory disease.

Evidence of Simian virus 40 (SV40) DNA sequences or gene products has been reported in a variety of organ systems in humans. However, the route of transmission and the significance of SV40 polyomavirus infection in human are unknown. The aim of study was to characterize the frequency of SV40 infection in immunocompetent and immunocompromised patients with respiratory diseases. Respiratory specimens from patients with respiratory tract illness obtained from nasopharyngeal aspirates (n = 280) were screened for SV40 polyomavirus using real-time PCR; coinfection with other viruses was examined. Positive results were confirmed with sequencing. Of the 280 samples analysed, 2 (0.71%) were positive for SV40. SV40 was identified in nasopharyngeal aspirate samples from children aged 8 and 14 months who were immunocompetent. Both patients had upper or lower respiratory tract infection. Coinfections with other viruses were found in 50% of the SV40 positive samples. The data suggest that SV40 can infect respiratory tract, that respiratory tract may represent a route of transmission or a site for virus persistence, and that with the high rate of co-infection, SV40 may not involved in respiratory diseases.

Synthesis and characterization of a novel magnetic porous carbon coated with poly(p-phenylenediamine) and its application for furfural preconcentration and determination in baby food and dry milk powder samples.

The aim of the current research is to develop a MSPE method for the determination of furfural in baby food and dry milk samples. In this regard, a novel magnetic porous carbon composite coated with poly(p-phenylenediamine) was fabricated, characterized, and then applied to the preconcentration/extraction of furfurals from baby food and dry milk powder samples. Initially, magnetic nanoparticles (Fe3O4) were synthesized, and then coated with a metal-organic framework layer named MIL-101(Fe). Afterward, the magnetic MIL-101(Fe) was subjected to calcination under a nitrogen atmosphere and magnetic porous carbon was achieved. Finally, a layer of poly(p-phenylenediamine) was coated on the magnetic porous carbon. The structure of the nanocomposite was investigated with various methods, including FT-IR spectroscopy, electron microcopies (SEM and TEM), VSM, and XRD. The fabricated nanocomposite was applied in magnetic solid-phase extraction of furfural and hydroxymethyl furfural and their determination with liquid chromatography. The effect of experimental variables was explored by using an experimental design approach. The LODs and linear range for the target furfurals were 1.0-2.0 µg kg-1 and 3.0-500 µg kg-1, respectively. The method's repeatability was explored using RSD values and was found to be in the range of 5.2-6.4% (one-day, n = 5) and 9.1-10.8% (day to day, n = 3). Eventually, this new method was employed for the extraction/quantification of target compounds in baby food and dry milk powder samples.

Transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to animals: an updated review.

COVID-19 caused by a novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) originated in Wuhan (Hubei province, China) during late 2019. It has spread across the globe affecting nearly 21 million people with a toll of 0.75 million deaths and restricting the movement of most of the world population during the past 6 months. COVID-19 became the leading health, economic, and humanitarian challenge of the twenty-first century. In addition to the considerable COVID-19 cases, hospitalizations, and deaths in humans, several cases of SARS-CoV-2 infections in animal hosts (dog, cat, tiger, lion, and mink) have been reported. Thus, the concern of pet owners is increasing. Moreover, the dynamics of the disease requires further explanation, mainly concerning the transmission of the virus from humans to animals and vice versa. Therefore, this study aimed to gather information about the reported cases of COVID-19 transmission in animals through a literary review of works published in scientific journals and perform genomic and phylogenetic analyses of SARS-CoV-2 isolated from animal hosts. Although many instances of transmission of the SARS-CoV-2 have been reported, caution and further studies are necessary to avoid the occurrence of maltreatment in animals, and to achieve a better understanding of the dynamics of the disease in the environment, humans, and animals. Future research in the animal-human interface can help formulate and implement preventive measures to combat the further transmission of COVID-19.

In vivo toxicological evaluation of graphene oxide nanoplatelets for clinical application.

BACKGROUND: Graphene is considered as a wonder material; it is the strongest material on the planet, super-elastic, and conductive. Its application in biomedicine is huge, with a multibillion-dollar industry, and will revolutionize the diagnostic and treatment of diseases. However, its safety and potential toxicity is the main challenge. METHODS: This study assessed the potential toxicity of graphene oxide nanoplatelets (GONs) in an in vivo animal model using systemic, hematological, biochemical, and histopathological examinations. Normal saline (control group) or GONs (3-6 layers, lateral dimension=5-10 µm, and thickness=0.8-2 nm) at dose rate of 50, 150, or 500 mg/kg were intraperitoneally injected into adult male Wistar rats (n=5) every 48 hours during 1 week to receive each animal a total of four doses. The animals were allowed 2 weeks to recover after the last dosing. Then, animals were killed and the blood was collected for hematological and biochemical analysis. The organs including the liver, kidney, spleen, lung, intestine, brain, and heart were harvested for histopathological evaluations. RESULTS: The results showed GONs prevented body weight gain in animals after 21 days, treated at 500 mg/kg, but not in the animals treated at 150 or 50 mg/kg GONs. The biochemical analysis showed a significant increase in total bilirubin, with a significant decrease in triglycerides and high-density lipoprotein in animals treated at 500 mg/kg. Nonetheless, other hematological and biochemical parameters remained statistically insignificant in all GONs treated animals. The most common histopathological findings in the visceral organs were granulomatous reaction with giant cell formation and accumulation of GONs in capsular regions. Also, small foci of neuronal degeneration and necrosis were the most outstanding findings in the brain, including the cerebellum. CONCLUSION: In conclusion, this study shows that GONs without functionalization are toxic. The future study is a comparison of the functionalized with non-functionalized GONs.

The Role of TNF-α in Aflatoxin B-1 Induced Hepatic Toxicity in Isolated Perfused Rat Liver Model.

Aflatoxin B-1 (AFB1) is one of the major mycotoxins causing food contamination. Previous studies have shown that AFB1 can induce carcinogenicity and toxic effects in the isolated perfused rat liver and these effects are associated with its metabolites and peroxidation activity. Here we surveyed whether these pathogenic effects of AFB1 are associated with TNF-α as an inflammatory cytokine in general liver damages. In this study, we used twenty male Wistar rats (250-300 g). Rats were divided into four groups. Control group was pre-treated with LPS and then perfused with KHBB. The second group was pretreated with PTX and LPS and then perfused with KHB. The third group was pre-treated with LPS and then perfused with AFB-1 and KHB. The last group was pretreated with LPS and PTX and then perfused with AFB1 and KHB. Results revealed that aflatoxin B1 significantly increased the enzyme activity of aminotransferase and levels of lipid peroxidation. Also, the levels of Glutathione decreased in the aflatoxin group significantly. TNF-α released in perfusate and increased in aflatoxin B1 group significantly and decreased in AFB-1+PTX. Exposure to Aflatoxin B1 may induce reactive oxygen species, so these species may induce overproduction of proinflammatory cytokines such as TNF-α and may cause more damage to hepatic cells.

Effects of Nano-zinc on Biochemical Parameters in Cadmium-Exposed Rats.

Cadmium (Cd) is a toxic environmental and occupational pollutant with reported toxic effects on the kidneys, liver, lungs, bones, and the immunity system. Based on its physicochemical similarity to cadmium, zinc (Zn) shows protective effects against cadmium toxicity and cadmium accumulation in the body. Nano-zinc and nano-zinc oxide (ZnO), recently used in foods and pharmaceutical products, can release a great amount of Zn2+ in their environment. This research was carried out to investigate the more potent properties of the metal zinc among sub-acute cadmium intoxicated rats. Seventy-five male Wistar rats were caged in 15 groups. Cadmium chloride (CdCl2) was used in drinking water to induce cadmium toxicity. Different sizes (15, 20, and 30 nm) and doses of nano-zinc particles (3, 10, 100 mg/kg body weight [bw]) were administered solely and simultaneously with CdCl2 (2-5 mg/kg bw) for 28 days. The experimental animals were decapitated, and the biochemical biomarkers (enzymatic and non-enzymatic) were determined in their serum after oral exposure to nano-zinc and cadmium. Statistical analysis was carried out with a one-way ANOVA and t test. P < 0.05 was considered as statistically significant. The haematocrit (HCT) significantly increased and blood coagulation time significantly reduced in the nano-zinc-treated rats. AST, ALT, triglyceride, total cholesterol, LDL, and free fatty acids increased significantly in the cadmium- and nano-zinc-treated rats compared with the controls. However, albumin, total protein, and HDLc significantly decreased in the cadmium- and nano-zinc-treated rats compared with the controls (P < 0.05). It seems that in the oral administration of nano-zinc, the smaller sizes with low doses and the larger sizes with high doses are more toxic than metallic zinc. In a few cases, an inverse dose-dependent relationship was seen as well. This research showed that in spite of larger sizes of zinc, smaller sizes of nano-zinc particles are not suitable for protection against cadmium intoxication.

Comparison of the Effects of Iron Oxide, as a New Form of Iron Supplement, and Ferrous Sulfate on the Blood Levels of Iron and Total Iron-Binding Globulin in the Rabbit.

Iron oxide is an important biological agent that has a key role in medical processes; however, the mechanism whereby it provides iron for human and animal cells and its biological uses remains unclear. We aimed to evaluate the effects of oral iron oxide on serum iron status and compare the results with those of iron sulfate as a reference salt. Fifteen adult rabbits were divided into 3 groups of 5 each: control group, iron sulfate group, and iron oxide group. The groups received doses of 3.3, 10, and 33 mg/kg in 3 experiments. Venous blood samples were obtained just before the oral administration of iron sulfate and iron oxide (3.3 mg/kg). More blood samples were taken 3 times at the time points of 1, 6, and 12 hours after the administration of the solutions. Serum was separated for the measurement of iron (Fe) and total iron-binding globulin (TIBG) with routine methods. One week later, the same experiment was repeated with 10 mg/kg of iron sulfate and iron oxide; and 1 week later after the second experiment, again the same experiment was repeated with 33 mg/kg of iron sulfate and iron oxide. The results showed that 33 mg/kg of iron sulfate 1 hour after treatment caused a significant difference in the Fe and TIBG levels between all the groups (P=0.014 for Fe and P=0.027 for TIBG). Our data showed that the absorption of iron oxide was similar to that of ferrous sulfate and in high doses was as useful as iron supplement.

Attenuation of Withdrawal Signs, Blood Cortisol, and Glucose Level with Various Dosage Regimens of Morphine after Precipitated Withdrawal Syndrome in Mice.

Morphine withdrawal usually results in unsuccessful outcomes. Despite partial benefits from alternative substances such as methadone, its use may not lead to the desired result due to the lack of mental tranquility during the withdrawal period. In this study, by means of an animal model, morphine itself was used to manage morphine dependence. Forty mice were divided into 5 groups, in which 4 groups became dependent by increasing daily doses of morphine for 7 days (15-45 mg/kg). Afterwards, the animals received morphine for 14 days by either of the following regimens: Once daily 45 mg/kg (positive controls)Increasing the interval (each time 6 hours longer than the previous interval)Irregular interval in every 36, 12 and 24 hours until the 21(th) day12, 24, 36 hours decreasing doses (each time 2.5 mg/kg less than the former dosage). Negative controls received saline solution only. On day 22, total withdrawal index (TWI) was determined by injecting 3 mg/kg of naloxone. Thereafter, blood samples were taken for the measurement of cortisol and glucose levels. TWI significantly decreased in all test groups in comparison with the positive control animals (P<0.001). Cortisol levels significantly decreased when either the dosage or the administration frequencies were decreased on a regular and gradual basis (P<0.005). Blood glucose levels significantly decreased in animals that received decreasing doses of morphine (P<0.005). This study suggests that no other measures may be required in clinical practice except for changing the dosage regimen of morphine for the cessation of self-administration.

Malathion induces anxiety in the male adult mouse.

INTRODUCTION: The cholinergic nervous system and acetylcholine esterase are involved in chronic intoxication with organophosphorous insecticides. The present study aims to investigate the influence of the chronic toxicity of these chemicals on behaviors related to anxiety, using the elevated plus maze (EPM), in the male adult mouse. MATERIAL AND METHODS: Either water or 1% concentration of malathion was applied dermally to the male adult mice (10 s, once daily for 28 days) and, on day 29, the EPM test was done. RESULTS: Time spent in the open arms (TSOA) in intoxicated animals was decreased by over 50% compared to the controls (p = 0.047). In contrast, time spent in closed arms was significantly higher in the malathion-exposed mice (p = 0.025). Percentage of open arm entries (OAE) was slightly smaller in the malathion-treated group in comparison to the control animals. Percentage of closed arm entries (CAE) in the treated group was slightly higher than the value in the control animals. CONCLUSIONS: The results showed that chronic toxicity of malathion may lead to an anxiety-like behavior in the animal model used in this study. It is difficult to extend these findings to clinical situations. However, more experimental work in different animal species as well as epidemiological studies in human subjects in this regard are highly recommended.

Amelioration of cardio-respiratory perturbations following Mesobuthus eupeus envenomation in anesthetized rabbits with commercial polyvalent F(ab')2 antivenom.

Immunotherapy is the only specific treatment for scorpion sting. In the present study, protective effects of polyvalent antivenom against hemodynamic disturbances, biomarkers (troponin T, creatinine kinase isoenzyme MB, Lactate dehydrogenase) changes, electrocardiogram abnormalities and histopathological complications in heart and lung induced by Mesobuthus eupeus scorpion venom was investigated in anesthetized rabbits. Twenty four rabbits were randomized into four equal groups: six rabbits in control group received 1 ml ultra-pure water subcutaneously (group 1). Group two received LD50 of venom (4.5 mg/kg). In the third and fourth groups, 5 ml of scorpion antivenom was administrated intravenously simultaneous with venom injection and 60 min following envenomation, respectively. Results of the present study indicate significant decrease in hemodynamic parameters following envenomation in the second group of animals. Venom injection caused edema, myocytolysis, coagulation necrosis, hemorrhage in heart as well as edema, hemorrhage and vascular thrombus in lungs. Although envenomed rabbits presented rises in LDH and TnT but no alteration in CK-MB was observed. Electrocardiogram monitoring of rabbits showed ST elevation and inverted T waves. Simultaneous administration of antivenom and venom prevented entirely the clinical signs, hemodynamic disturbances, markers changes, ECG abnormalities and histopathological damages. Delayed immunotherapy gradually ameliorated clinical signs, hemodynamic disturbances and markers changes related to envenomation. Histopathological evaluation showed slight alterations such as mild myocytolysis in heart and mild edema in lung following delayed immunotherapy. In conclusion, scorpion antivenom administration has preventive, neutralizing and curative properties for M. eupeus scorpion envenomation, if it would be applied at optimum time, dose and route.

Development of DNA-designed avian IgY antibodies for quantitative determination of bovine interferon-gamma.

Interferon-gamma (IFN-γ), a cytokine produced by sensitized T lymphocytes, is one of the key elements in defining T helper 1 lymphocyte immune responses. Quantitative evaluation of IFN-γ expression could provide an important analytical tool for measurement of cell-mediated immunity and investigating immune responses to infectious diseases. Method of DNA-designed avian IgY antibodies was used for production of monospecific polyclonal antibodies that allows quantification of the recombinant bovine IFN-γ protein. IFN-γ cDNA was subcloned and expressed in mammalian expression plasmid (pcDNA3.1(+)) under the control of the human cytomegalovirus promoter. Chickens were immunized by plasmid DNA, and egg yolk antibodies extracted from eggs were collected after immunization. IgY-specific antibodies were evaluated by an antigen capture enzyme-linked immunosorbent assay (ELISA) using recombinant IFN-γ. Based on the results, developed bovine IFN-γ capture ELISA could detect up to 1 ng/ml of IFN-γ by 64-fold diluted IgY. Monospecific anti-bovine IFN-γ antibodies generated in chickens are useful for quantifying different concentrations of recombinant bovine IFN-γ, which is expressed in cell culture.

A novel molecular assay to discriminate transcriptional effects caused by xenoestrogens.

A phenotypic definition of the term estrogen has become increasingly problematic due to the multiple modes of estrogen action which can now be defined by differing nuclear and membrane receptors for the classic ligand, 17beta-estradiol, and by the multiple signalling pathways that are consequently addressed. This has led to the term xenoestrogen being largely determined by whatever assay system is used for its definition. Here we describe a novel and simple matrix for a transfection system using MBA-MD231 and MCF-7 breast cancer cells as hosts. This matrix is able to vary the type of nuclear estrogen receptor used, and by varying the promoter-reporter construct between one using a classic estrogen response element (ERE) enhancer, and one using an enhancer element derived from the bovine oxytocin gene promoter binding an orphan nuclear receptor, direct classical effects can be neatly discriminated from non-classical and non-genomic actions of test substances. This assay matrix has been used to examine a selection of phytoestrogens and xenobiotics, thereby providing new information on the mechanism of action of some of these substances in breast cancer cells.