Interaction of lithium and sleep deprivation on memory performance and anxiety-like behavior in male Wistar rats.

Sleep plays a crucial role in the modulation of physiological and cognitive functions. Many studies have reported the impairment effect of sleep deprivation (SD) on cognitive functions such as learning and memory. On the other hand, lithium as one of the oldest drugs used for the treatment of psychiatric disorders, affects cognitive functions and mood state. In this study, we aimed to assess the effect of lithium, SD (for 24 h), and the interaction effect of SD and lithium, on memory function and anxiety-like behavior. The water box, the shuttle box, elevated plus maze, and the three-chamber paradigm test were used to evaluate rat's behavior. Also, lithium was injected intraperitoneal at the doses of 10 and 20 mg/kg, for three consecutive days. The results showed that SD impaired passive avoidance memory and social interaction memory, and decreased anxiety-like behavior. Lithium also impaired passive avoidance memory and induced an anxiolytic effect, while it improved social interaction memory and reversed the impairment effect of SD on social interaction memory. In conclusion, we suggested that interaction effect of SD and lithium on the function of brain-derived neurotrophic factor (BDNF) and glycogen synthase kinase3-ß (GSK3-ß) may be involved in the modulation of cognitive functions. As a limitation of this research, it was declared that we did not evaluate the function of GSK3-ß and BDNF in the brain of rats, especially in the hippocampus. We suggested conducting more studies focusing on the interaction of SD and lithium on the function of BDNF and GSK3-ß, and on different cognitive functions.

Rapid and Sensitive Detection of Staphylococcal Enterotoxin B by Recombinant Nanobody Using Phage Display Technology.

Staphylococcal enterotoxin B, from Staphylococcus aureus (S. aureus), is one of the most potent bacterial superantigens with profound toxic effects on the immune system. It is associated with food poisoning, toxic shock, atopic dermatitis, asthma, and nasal polyps in humans. The current diagnostic methods for staphylococcal enterotoxin are mainly based on traditional monoclonal antibodies which hardly meet the requirements for clinical applications, and hybridoma clones lose their ability to secrete antibodies during time. The present study investigates the development of a novel, highly specific, low-cost, and sensitive nanobody capable of being used in immunoassays for Staphylococcal enterotoxin B (SEB) detection in suspicious foods. For this purpose, Camelus dromedarius was immunized against SEB toxin. After obtaining acceptable titration, a high-quality phage display nanobody library (4 × 1010 PFU/ml) was constructed. High-affinity SEB-specific nanobodies were retrieved from constructed libraries. After phage rescue and five round of biopanning, clone screening was performed by phage ELISA. Recombinant nanobodies which were expressed from C7 and C21 clone showed the highest affinity for SEB. The presence of high quality and pure nanobody band at ~ 15 kDa was confirmed by SDS-PAGE and western blotting. The affinity constant which was measured by ELISA was calculated to be around 10-9 M. The results suggest that the proposed detection method by nanobodies is an alternative diagnostic tool enabling a rapid, inexpensive, and specific detection of the SEB.

Comparison of different solvents for extraction of polyhydroxybutyrate from Cupriavidus necator.

Polyhydroxybutyrate (PHBs) have attracted much attention due to their biodegradability and biocompatibility properties. The main drawback to the commercial production of them is their high cost. The recovery of PHB from bacterial cytoplasm significantly increases total processing costs. Efficient, economical, and environment-friendly extraction of PHB from cells is required for its industrial production. In the present study, several nonhalogenated organic solvents (ethylene carbonate, dimethyl sulfoxide, dimethyl formamide, hexane, propanol, methanol, and acetic acid) were examined for their efficacy regarding recovery at different temperatures from culture broth containing Cupriavidus necator cells. The highest recovery percentage (98.6%) and product purity (up to 98%) were seen to be those of ethylene carbonate-assisted extraction at 150°C within 60 min of incubation time. Average molecular weight of the recovered PHB (1.3 × 106) was not significantly affected by the extraction solvent and conditions. The melting point of PHB extracted using ethylene carbonate was measured to be 176.2°C with an enthalpy of fusion of 16.8% and the corresponding degree of crystallinity of 59.2%. NMR and GC analyses confirmed that the extracted biopolymer was PHB. The presented strategy can help researchers to reduce the cost to obtain the final product.

Evaluation of the hemocompatibility of RADA 16-I peptide.

RADA 16-I is an ionic self-assembling peptide that can form macroscopic scaffolds through ß-sheet structures which are used in favor of cell growth and tissue engineering. This peptide has also the ability to stop bleeding effectively and quickly (∼20 seconds) when applied directly to the injuries. This study is focused on coagulation process, platelet aggregation, C3 and C4 concentrations, CBC counting, hemolysis, and white blood cell morphology tests to analyze hemocompatibility of RADA 16-I at different concentrations - 0.1, 0.2, 0.3 and 0.5%. According to the results, RADA 16-I hydrogel decreased the number of blood cells, slightly increased clot formation time and platelet aggregation, and yielded negligible hemolysis and only small changes in C3 and C4 concentrations and white blood cell morphology. All by all, the in vitro tests of hemocompatibility showed no perturbation in the blood composition when the peptides were in contact with the blood. The observed rapid hemostasis might be a result of increasing local concentrations of molecules involved in the formation of clot near the peptide hydrogel, thereby making a barrier which ended up with complete hemostasis. In conclusion, our experiments strongly supported further development of biomaterials based on RADA 16-I peptide.

Evaluation of Apoptosis in Multipotent Hematopoietic Cells of Bone Marrow by Anthracycline Antibiotics.

Anthracycline antibiotics are potent anticancer drugs widely used in the treatment of solid tumors and hematological malignancies. Because of their extensive clinical use and their toxic effect on normal cells, in the present study the effect of these drugs on multipotent hematopoietic bone marrow cells was investigated employing, viability tests, PARP cleavage, Hoechst 33258 staining, DNA fragmentation and superoxide anion production techniques. The results revealed that daunorubicin and doxorubicin exhibited time and dose dependent cytotoxicity against the cells and upon increasing the drugs concentrations, apoptosis was occurred after 4 h of incubation and at low concentration of the drugs. The cleavage of poly ADP-ribose polymerase (PARP) demonstrated by daunorubicin and doxorubicin treatment of the cells, suggest that the apoptotic process is PARP dependent. The drugs induced DNA fragmentation and also anion superoxide production was increased upon rising drugs concentrations. From the results it is concluded that anthracycline antibiotics represent cytotoxic effect on hematopoietic progenitor/stem cells of bone marrow, inducing apoptosis and in this process toxicity of daunorubicin is more pronounced compared to doxorubicin.

An efficient method for the application of PHA-poor solvents to extract polyhydroxybutyrate from Cupriavidus necator.

There are many published studies presenting ethanol and acetone as PHAs-poor solvents, where these two solvents are shown to dissolve <2% (w/v) of PHAs at low temperatures. In this study, the suitability of ethanol and acetone for the recovery of PHB at different temperatures (from room temperature to near boiling point) in Cupriavidus necator was investigated. Experiments were performed using response surface methodology to examine the effects of different temperatures and heating incubation times on recovery percentage using the two solvents. The highest recovery percentage (92.3%) and product purity (up to 99%) were obtained with ethanol-assisted extraction at 76°C for 32 min of incubation time. Under these conditions the extracted PHB exhibited a molecular mass of 1.2 × 106 . The present strategy showed that at temperatures near its boiling point, ethanol, as a nonhalogenated solvent, represents a good alternative to halogenated solvents, like chloroform, when PHB recovery is concerned. DSC analysis showed good thermal properties for ethanol- and acetone-extracted biopolymers. GC and 1 H NMR analysis confirmed the extracted biopolymer to be polyhydroxybutyrate of good purity. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1480-1486, 2016.

Effective enhancement of hydroxyvalerate content of PHBV in Cupriavidus necator and its characterization.

This study investigated the effects of using a combination of substrates (fructose, propanol, citric acid, acetic acid, propionic acid and beef extract) on the synthesis of poly (3-hydroxybutyrate-co-3-hydroxyvalerate), which is also known as PHBV. The results of batch PHBV optimization by RSM, indicated that PHB was synthesized only when fructose was used as the sole carbon source. Fructose in combination with propanol and beef extract, exhibited better PHBV production performance, as compared to that of propionic acid which was used in previous studies. GC, (1)H and (13)C NMR spectroscopy revealed that the best copolymer composition was 54.1 mol% of 3-hydroxyvalerate (3HV) which was obtained in a basal medium combined with 0.3% (v/v) propanol and 0.0172% (w/v) beef extract. This is the first report on the production of PHBV with HV content of 54.1% by a wild-type Cupriavidus necator. The Fourier Transform Infrared Spectroscopy (FTIR) spectra of the PHAs indicated absorption bands at 1723, 1274, 1373, 1453, 2932cm(-1) corresponding to CO, CO stretching, CH3, -CH2 and -CH groups, respectively. This study showed that the recovered PHBV biopolymer from the optimized culture to have a melting temperature of 103.6°C, melting enthalpy of 55.67J/g and crystallization temperature of 76.1°C.

High production of bacteriorhodopsin from wild type Halobacterium salinarum.

Bacteriorhodopsin (bR) is a trans-membrane proton pump found in the purple membrane of Halobacterium salinarum. This protein has high photochemical and photoelectric conversion efficiency and thermal stability, allowing it to withstand high temperatures, high salinity, and nutritionally-limited environments. The ability of this protein to convert light energy into chemical energy has applications that are mainly therapeutic/diagnostic and research-oriented. There is increasing demand for bacteriorhodopsin production in different fields. The present study maximized bacteriorhodopsin production using H. salinarum. The physical parameters of illumination, agitation speed, temperature, and nitrogen source were studied using a fractional factorial design to determine the optimal levels of each. The most suitable nitrogen source was determined to be peptone from meat. The optimal temperature was 39 °C, agitation speed was 150 rpm, and light intensity was 6300 lux for bR production. Under these conditions, the maximum bR yield was 196 mg/l, which is about 4.23 fold greater than those obtained with basal medium. The proposed strategies could be used for bR production using this archaeobacterium; the results are the highest reported thus far from a batch culture of H. salinarum.

An Environmentally Friendly and Efficient Method for Extraction of PHB Biopolymer with Non-Halogenated Solvents.

The present study developed an efficient and environmentally friendly method for recovering polyhydroxybutyrate (PHB) from Cupriavidus necator. Several non-halogenated solvents were tested and it was found that butyl acetate and ethyl acetate are powerful solvents for the biopolymer. Testing was performed to examine the effects of temperature (25 °C until temperature below solvent boiling points) and heating incubation time (0-60 min) on the two solvents. Butyl acetate had a higher recovery level (96%) and product purity (up to 99%) than ethyl acetate at 103 °C and a heating incubation time of 30 min. Under these conditions, PHB recorded the highest molecular weight of 1.4 × 10(6) compared with the standard procedure (i.e., recovery using chloroform). The proposed strategy showed that butyl acetate is a good alternative to halogenated solvents such as chloroform for recovery of PHB.

An in vitro study on the effect of vinca alkaloid, vinorelbine, on chromatin histone, HMGB proteins and induction of apoptosis in mice non-adherent bone marrow cells.

CONTEXT: Vinoreline is a vinca alkaloid anticancer drug widely used in cancer therapy. Drugs are not target specific, therefore might affect normal tissues/cells, in which bone marrow is the important one. OBJECTIVE: To elucidate the cytotoxic and genotoxic effect of vinca alkaloid anti cancer drug, vinorelbine, on mice non-adherent bone marrow cells in vitro. MATERIALS AND METHODS: Non-adherent bone marrow cells were isolated and exposed to various concentrations (0-160 µg/ml) for 4 h at 23 °C. The chromatin proteins were analyzed by SDS PAGE and western blot. Fluorescent dye staining of the cells, anion superoxide and DNA fragmentations assays were also employed. RESULT: The results from MTT and trypan blue exclusion assays represented reduction of the cells viability. Extractability of histones and HMG proteins contrasted with difficulty as their content was decreased on SDS-gel upon increasing drug concentration as western blots confirmed it. The amount of degradation form of PARP (89 KD) increased significantly in a dose dependent manner. Increase in anion superoxide production and DNA fragmentation together with cytological detection of chromatin condensation and cellular damage upon exposure of the cells to vinorelbine were indicative of apoptosis induction in these normal cells. CONCLUSION: Vinorelbine is genotoxic in non-adherent bone marrow cells as affects chromatin components, DNA, histone and HMGB1 proteins and induces apoptosis.

Evidence for the genotoxic effect of daunomycin in multipotent hematopoietic cells of mouse bone marrow: chromatin proteins analysis.

Severe bone marrow suppression and anemia are the main side effects of using chemotherapeutic agents like daunomycin. In this study the effect of daunomycin on viability and chromosomal proteins of multipotent hematopoietic cells (MHCs) of mouse bone marrow was investigated. The cells were separated from mature cells according to their adherence, incubated in the absence and presence of various concentrations of daunomycin and viability was determined by trypan blue exclusion and MTT assay. The histones and high mobility group (HMG) proteins were extracted by acid and salt, respectively and analyzed on SDS-PAGE and immunoblot. The results revealed that daunomycin exhibited time and dose dependent cytotoxicity on the cells and upon increasing the concentrations of drug, the extractability of histone and HMG proteins decreased. Agarose gel electrophoresis showed aggregation of DNA in the presence of drug. Flow cytometry analysis suggested that there are differences in type and amount of histones H2A and H2B of these cells in comparison with thymus histones. Moreover histone H3 was not subjected to trimethylation at lysine 9 and daunomycin decreased H3K9 dimethylation and H3K9 acetylation in these cells in a dose-dependent manner. From these results it is concluded that the binding of daunomycin to chromatin precedes chromatin of multipotent hematopoietic cells of bone marrow into compaction/aggregation and beside DNA, histones and HMG proteins also play an important role in this process.