Transplanting P75-Suppressed Bone Marrow Stromal Cells Promotes Functional Behavior in a Rat Model of Spinal Cord Injury

Background: Bone marrow stromal cells (BMSC) have been successfully employed for movement deficit recovery in spinal cord injury (SCI) rat models. One of the unsettled problems in cell transplantation is the relative high proportion of cell death, specifically after neural differentiation. According to our previous studies, p75 receptor, known as the death receptor, is only expressed in BMSC in a time window of 6-12 hours following neural induction. Moreover, we have recently reported a decreased level of apoptosis in p75-suppressed BMSC in vitro. Therefore, our objective in this research was to explore the functional effects of transplanting p75:siRNA expressing BMSC in SCI rats. Methods: Laminectomy was performed at L1 vertebra level to expose spinal cord for contusion using weight-drop method. PBS-treated SCI rats (group one) were used as negative controls, in which cavitations were observed 10 weeks after SCI. pRNA-U6.1/Hygro- (group two, as a mock) and pRNA-U6.1/Hygro-p75 shRNA- (group three) transfected BMSC were labeled with a fluorescent dye, CM-DiI, and grafted into the lesion site 7 days after surgery. The Basso-Beattie-Bresnehan locomotor rating scale was performed weekly for 10 weeks. Results: There was a significant difference (P≤0.05) between all groups of treated rats regarding functional recovery. Specifically, the discrepancy among p75 siRNA and mock-transfected BMSC was statistically significant. P75 siRNA BMSC also revealed a higher level of in vivo survival compared to the mock BMSC. Conclusion: Our data suggest that genetically modified BMSC that express p75:siRNA could be a more suitable source of cells for treatment of SCI. Key Words: Spinal cord injury (SCI), Apoptosis, Bone marrow stromal cells (BMSC)     

Extraction, characterization, and nematicidal activity of chitin and chitosan derived from shrimp shell wastes

Chitin has been chemically extracted from the Egyptian shrimp shell waste. The obtained chitin was transformed into the more soluble chitosan. Chitin and chitosan were characterized using Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectrometry. These products were also characterized by their nematicidal potential against the root-knot nematode, Meloidogyne incognita, infecting tomato in a glasshouse. Chitin or chitosan was incorporated into the soil at the rate of 1, 3, 5, and 10 g/kg, and their nematicidal activity was compared with that of the synthetic nematicide oxamyl at the rate of 0.01 g a.i./kg. The effects of the treatments on the growth of tomato were also examined. The obtained results show that chitin and chitosan reduced tomato root galls and J₂ of M. incognita in the soil in a dose-dependent manner. Chitosan was more effective in the reduction of galls and J₂ in the soil than chitin. However, the efficacy of chitin and chitosan in reducing the number of J₂ in the soil was less than that of oxamyl. As compared to the untreated inoculated plants, the length of shoots and roots was significantly (p ≤ 0.05) increased in soil amended with either chitin or chitosan, whereas both treatments did not significantly (p > 0.05) differ with respect to the inoculated control on the effect on the dry weight of shoot and root systems. Furthermore, neither chitin nor chitosan treatments at the tested rates were phytotoxic to tomato plants.     

The effects and roles of PVP on the phase composition, morphology and magnetic properties of cobalt ferrite nanoparticles prepared by thermal treatment method

Cobalt ferrite nanocrystals were prepared from an aqueous solution containing metal nitrates and various of concentrations of poly(vinyl pyrrolidone), followed by calcinations temperature. X-ray diffraction (XRD) analysis was performed to determine the degree of crystallinity of the ferrite nanoparticles. By transmission electron microscopy (TEM), the morphology and average particle size of the cobalt ferrite nanoparticles were evaluated which had good agreement with XRD results. Fourier transform infrared spectroscopy (FT-IR) suggested the presence of metal oxide bands in all samples as well as the effective elimination of organic constituents after calcinations. Vibrating sample magnetometer (VSM), was utilized to evaluate the magnetic properties of the cobalt ferrite nanoparticles.     

The role of Toll‐like Receptors in innate immunity and infectious diseases of teleost

The immune defence mechanism depends mainly on germ‐line encoded pattern‐recognition receptors (PRRs). These PRRs respond to many exogenous pathogens and/or endogenous serious signals, by recognizing some highly conserved structures such as pathogen‐associated molecular patterns (PAMPs) and danger/damage associated molecular patterns (DAMPs). Till date, the most studied PRRs are Toll‐like receptors (TLRs). Upon activation of TLRs, there is production of inflammatory cytokines and type I interferons (IFNs) via myeloid differentiation primary response gene 88 (MyD88)‐dependent or ‐independent signalling, respectively, modulating innate and adaptive immunity, as well as inflammatory responses. In fish species studied to date, there are more than 17 TLRs that are identified with some showing homology to mammals, and some are unique for teleost. In the present review, more light are to be shed on the classification, structure and specific ligands of TLRs, with focuses on their signal pathways and different biological activities. Studies of TLRs and their role in the innate immune will potentially have implications for the prevention and treatment of fish diseases.     

Evaluating interspecific wheat hybrids based on heat and drought stress tolerance

Three durum and three bread wheat genotypes were crossed to produce three tetraploid, three hexaploid and nine interspecific (pentaploid) F₁ hybrids. All genotypes were evaluated for heat tolerance in the field and for drought using polyethylene glycol in vitro. Chromosome numbers and meiotic behavior in pentaploid F₁ hybrids (2n=5x=35, genomes AABBD) were confirmed. Heat stress significantly reduced grain yield/plant and 1000-kernel weight (1000-KW), while grain protein content (GPC) was increased. Drought caused a significant reduction in root length, shoot length and seedling fresh weight, whereas root/shoot ratio was increased. P₃ (durum), P₄ (bread) and their pentaploid F₁ hybrid could be considered as the most heat-tolerant genotypes. However, P₂ (durum), P₆ (bread) and their F1 were most tolerant to drought. The addition of a D genome single dose into pentaploid F1 hybrids obviously reduced grain yield/plant, 1000-KW and seedling traits, however GPC was increased. Moderate to high broad-sense heritability and genetic advance were obtained for the most investigated traits. Grain yield/plant was strongly positively correlated with stress tolerance index (STI), yield index (YI), mean productivity (MP), geometric mean productivity (GMP) and harmonic mean (HM) under heat stress and with root length under drought condition, suggesting that STI, YI, MP, GMP and HM are powerful indices for heat tolerance, while root length is most effective for drought. Successful interspecific hybridization obtained in the study is only an initial step for desired genes introgression. Successive progenies are going to be evaluated for further genetic studies aiming at improving abiotic stress tolerance in wheat.     

Effect of genotype and growing season on nitrate accumulation and expression patterns of nitrate transporter genes in potato (Solanum tuberosum L.)

Nitrate (NO₃^?) is the most important source of mineral nitrogen (N) affecting the vegetative growth, the tuber yield and the quality of potato (Solanum tuberosum L.). The screening of 16 potato cultivars for low nitrate content was reported and the effects of genotype and experimental factors on nitrate accumulation were studied. Both genotype and growing season had a highly significant effect on dry weight, number of tubers/plant, yield and nitrate accumulation in potato which varied significantly from one genotype to another. Furthermore, nitrate accumulation varies with the season, being higher in the first season (autumn) than in the second season (spring), for all the genotypes. The cultivars Safrane and Universa followed by Actrice and Alaska produced the highest tuber yield/plant during the first and second season, respectively, accumulating higher quantities of nitrate compared to other cultivars. Ponomera cultivar was of interest from a breeding perspective due to both combining best tuber yield/plant and low nitrate accumulation for both growing season. Expression pattern of nitrate transporter genes (StNRT1.2, StNRT1.5 and StNRT2.1) has been reported. Thus, these genes were differentially expressed which could be explained in part by the difference in nitrate accumulation rate between the studied cultivars. The results showed that high accumulator genotype got significant higher expression level of StNRT1.2, StNRT1.5 or StNRT2.1 which lead to the higher nitrate uptake rate and consequently higher nitrate accumulation than low accumulator genotype.     

Evaluation of drought and heat tolerance in wheat based on seedling traits and molecular analysis

Polyethylene glycol and cell membrane stability (CMS) assay were used to evaluate drought and heat tolerance, respectively, among 14 wheat lines based on seedling traits and molecular analysis. Significant variation was evidenced for all the investigated seedling traits. Different levels of heritability and genetic advance were found among the tested traits, indicating whether the trait is controlled by additive or non-additive gene action. Drought caused a significant reduction in root and shoot lengths. However, root/shoot ratio under drought stress was increased. Root length showed a highly significant negative correlation with drought susceptibility index (DSI) under drought conditions. Cluster analysis based on seedling traits separated lines mainly by DSI and CMS. Some lines showed drought and heat tolerance by exhibiting a low DSI with high CMS. Sequence-related amplified polymorphism (SRAP) generated a total of 135 bands, with a level of polymorphism ranging from 30 to 86% among the tested lines. SRAP showed its efficiency in discriminating wheat genotypes by gathering all high-DSIlines in one sub-cluster and generating 10 and 3 unique and specific bands for high-DSI-lines and low-DSI-lines, respectively. These bands could be used for further work as SRAP markers associated with drought tolerance in wheat.     

Pathology and molecular diagnosis of paratuberculosis of camels

Camels are the prime source of meat and milk in many desert regions of the world including Saudi Arabia. Paratuberculosis of camels, locally called Silag, is a serious and invariably fatal disease in the Arabian camel. Six camels were used in this study. Five camels with clinical paratuberculosis were used to study the pathology of the disease and confirm its aetiology. The sixth camel was clinically healthy and used as a control. The camels were examined clinically and bled for haematological and blood chemistry analysis. They were then humanely killed with a high intravenous dose of thiopental sodium (10 mg/kg) for pathological studies as well as obtaining tissues for microbiological and molecular studies. The clinical signs of the disease were emaciation, diarrhoea, alopecia, wry neck and pale mucous membranes. Laboratory diagnosis showed reduced haemoglobin concentration, low haematocrit and high activity of the serum enzyme alanine aminotransferase. Serum creatinine concentration was normal. These results indicated the infected camels were anaemic and the function of their livers was affected. Postmortem examination showed thickened and corrugated intestinal mucosa, enlarged granulomatous mesenteric lymph nodes, miliary and diffuse granulomas in the liver (in four camels), generalized lymph node granulomas (in one camel), splenic granuloma (in one camel) and mediastinal lymph node granuloma (in two camels). Histopathological examination showed diffuse infiltration of macrophages in all organs showing lesions. Ziehl–Neelsen staining of tissue scraping and tissue sections showed masses of acid fast bacilli, except for the spleen. Infection with Mycobacterium avium subsp. paratuberculosis was confirmed by PCR by targeting the IS900 gene.     

Astaxanthin Mitigates Thiacloprid-Induced Liver Injury and Immunotoxicity in Male Rats

Thiacloprid (TCP) is a widely used neonicotinoid insecticide with a probable toxic hazard to animals and human beings. This hazard has intensified the demand for natural compounds to alleviate the expected toxic insults. This study aimed at determining whether astaxanthin (ASX) could mitigate the hepatotoxic effect of TCP and diminish its suppressive effect on immune responses in rats. Animals received TCP by gavage at 62.1 mg/kg (1/10th LD₅₀) with or without ASX at 40 mg/kg for 60 days. Intoxicated rats showed modulation of serum transaminases and protein profiles. The hemagglutination antibody titer to sheep red blood cells (SRBC) and the number of plaque-forming cells in the spleen were reduced. The cell-mediated immunity and phagocytosis were suppressed, while serum interleukins IL-1β, IL-6, and IL-10 were elevated. Additionally, malondialdehyde, nitric oxide, and 8-hydroxy-2′-deoxyguanosine levels were increased in the liver, spleen, and thymus, with depletion of glutathione and suppression of superoxide dismutase and catalase activities. The expressions of inducible nitric oxide synthase and the high mobility group box protein 1 genes were upregulated with histomorphological alterations in the aforementioned organs. Cotreatment with ASX markedly ameliorated the toxic effects of TCP, and all markers showed a regression trend towards control values. Collectively, our data suggest that the protective effects of ASX on the liver and immune system of TCP-treated animals depend upon improving the antioxidant status and relieving the inflammatory response, and thus it may be used as a promising therapeutic agent to provide superior hepato- and immunoprotection.