A Signature of Three microRNAs Is a Potential Diagnostic Biomarker for Glioblastoma

Background:Glioblastoma is the most common primary malignant neoplasm of the central nervous system. Despite progress in diagnosis and treatment, glioblastoma still has a poor prognosis. This study aimed to examine whether a signature of three candidate miRNAs (i.e. hsa-let-7c-5p, hsa-miR-206-5p, and hsa-miR-1909-5p) can be used as a diagnostic biomarker for distinguishing glioblastoma from healthy brain tissues. Methods:In this study, 50 FFPE glioblastoma tissue samples and 50 healthy tissue samples adjacent to tumor were included. The expression of each candidate miRNA (i.e. hsa-let-7c-5p, hsa-miR-206-5p, and hsa-miR-1909-5p) was measured using RT-qPCR. To show the roles of each miRNA and their biological effects on glioblastoma development and clinicopathological characteristics, in silico tools were used. ROC curves were performed to assess the diagnostic accuracy of each miRNA. Results:Based on the results, hsa-let-7c-5p and hsa-miR-206-5p were downregulated, while hsa-miR-1909-5p was upregulated in glioblastoma tumors compared to healthy samples. No association was detected between the expression of each candidate miRNA and sex. Except for hsa-let-7c-5p, other miRNAs did not correlate with age status. ROC curve analysis indicated that the signature of candidate miRNAs is a potential biomarker distinguishing between glioblastoma and healthy samples. Only hsa-miR-206-5p suggested the association with poor prognosis in glioblastoma patients. Conclusion:Our findings revealed that the signature of three miRNAs is capable of distinguishing glioblastoma tumor and healthy tissues. These results are beneficial for the clinical management of glioblastoma patients.Key Words: Inflammation, Gastric cancer, Gene expression, Pattern recognition receptors, RNA-seq     

Origin and genetic diversity of Tunisian grapes as revealed by microsatellite markers

Ten SSR loci, previously developed for grapevine, were analyzed to evaluate the genetic variability, cultivar relatedness, and parentage in a collection of 61 autochthonous Vitis vinifera cultivars from Tunisia.The number of alleles per locus ranged from 6 to 11, while the number of genotype patterns varied between 10 and 21. The expected heterozygosity varied between 0.621 and 0.855 and the observed heterozygosity was higher than 0.9 at 4 loci (VVMD28, VVMD5, VVIP31 and VVS2) indicating that the SSRs were highly informative.Cluster analysis using unweighted pair group method with arithmetic averaging (UPGMA) suggested 14 groups among studied cultivars and 53 grapevine denominations out of 61 were unequivocally distinguished, with all accessions showing at least one-specific combination of alleles.On the other hand, in order to overcome the existing confusion in Tunisian grapevine nomenclature, of the analyzed homonymous pairs of cultivars, only ‘Balta 2’ and ‘Balta 3’ have shown identical allelic profiles, consistent with their being the same genotype. Hence, nomenclature distinction is meaningless and only one denomination should be retained.Due to the high overall power of exclusion (Q) (greater than 99.99%) and to the absence of null alleles, the set of microsatellite loci used is appropriate to determine parentage in Tunisian grapevines beyond any reasonable doubt. The analysis of fingerprints indicated that the Tunisian grape vines have evolved through out crossing between five possible parents: Balta 1, Beldi Baddar, Beldi Rafraf, Beldi Local Rafraf and Khedhiri 3.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

Genetic structure and differentiation among grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) accessions from Maghreb region

Three gene pools representative of Vitis vinifera L. subsp. vinifera (=subsp. sativa Beger) growing in the Maghreb regions (North Africa) from Tunisia (44), Algeria (31) and Morocco (18) and 16 wild grape accessions (Vitis vinifera L. subsp. sylvestris (Gmelin) Beger) from Tunisia were analysed for genetic diversity and differentiation at twenty nuclear microsatellites markers distributed throughout the 19 grape chromosomes. 203 alleles with a mean number of 10.15 alleles per locus were observed in a total of 109 accessions. Genetic diversities were high in all populations with values ranging from 0.6775 (Moroccan cultivars) to 0.7254 (Tunisian cultivars). F _st pairwise values between cultivated grapevine populations were low but found to be significantly different from zero. High F _st pairwise values were shown between wild and cultivated compartments. Two parent offspring relationships, two synonyms and two clones of the same cultivar were detected. The rate of gene flow caused by vegetative dissemination of cultivated grapevine plants was not sufficient to genetically homogenise the pools of cultivars grown in different regions. The Neighbour Joining cluster analysis showed a clear separation according to geographical origins for the cultivated grapevines gene pools and revealed a high dissimilarity between cultivated and wild grapevine. However, three cultivars (Plant d’Ouchtata 1, Plant de Tabarka 3 and Plant d’Ouchtata 3) are very close to wild accessions and may result from a hybridisation between cultivated and wild accessions. The high level of differentiation between cultivated and wild accessions indicates that the cultivated accessions do not derive directly from local wild populations but could mostly correspond to imported materials introduced from others regions during historical times or derived from crossing between them.     

Quercetin attenuates H2O2‐induced toxicity of rooster semen during liquid storage at 4°C

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H₂O₂ (40 μM) and combination groups which incubated with constant dose of H₂O₂ (40 μM) plus various levels of quercetin (20, 40 and 80 μM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 μmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 μmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 μmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co‐administration of quercetin (especially at 40 and 80 μM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide‐mediated damage during cold liquid storage of rooster semen.     

Mucoadhesive Chitosan Electrospun Nanofibers Containing Tetracycline and Triamcinolone as a Drug Delivery System

The mucoadhesive Chitosan (CS) nanofibers as a drug delivery system were developed. Chitosan was modified via the immobilization of thiol groups from L-cysteine as a mucoadhesive reagent. The mucoadhesive properties of the chitosan nanofibers were evaluated by tensiometer set and via tensile studies. Drug and mucoadhesive agent loading lead to decrease diameters and increased porous of nanofibers. The release of Tetracycline (Tet) and Triamcinolone (Tri) were increased with increasing immersion time and it became constant at long immersion times. Mucoadhesion studies were done at pH 2–7 and in pH 6 maximum mucoadhesive properties observed. Release studies demonstrated a sustained release of both drug continued up to 48 hours. Microbial studies were performed on the nanofibers. The drug delivery system represented a novel tool for improve the therapeutic efficacy of various drugs that are poorly absorbed from the gastrointestinal tract. Also it is an efficient system for treatment of oral ulceration.     

Response of European seabass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) to canola oil diets: effect on growth performance,fish health and liver and intestine histomorphology

This study was undertaken to assess the effects of feeding European seabass (Dicentrarchus labrax) canola oil-added diets on growth, health status and liver and intestine histomorphology. Seabass (56.18 ± 0.16 g initial body weight) were fed one of three fish meal-based diets with ~48 % crude protein and ~16.0 % lipid, combining fish oil (FO) and canola oil (CO) for 12 weeks. The diets contained: zero (control, CTRL), 45 (CO50) or 63 (CO70) g CO kg^?1 assigned in triplicates to three dietary groups. The results indicated that neither dietary oil type (FO or CO) nor CO level adversely affected (P > 0.05) the growth, feed utilization or major blood constituents’ composition as an indicator of the overall health status of fish. Despite the CO diets influence on head kidney macrophage activity being unappreciable (P > 0.05), there was a reducing trend with an increase in CO level incorporation. The CO70 diet induced a minor fat infiltration in hepatocytes and leucocytes infiltration, hyperplasia of the basal nuclei and supranuclear vacuolization of the enterocytes of the distal intestine. The present observations suggest that it is possible to incorporate up to 63 g CO Kg^?1 in the feed for European seabass juveniles without major negative effects on growth, health status or liver and intestinal histomorphology.     

The Effects of Estrogen Receptors' Antagonist on Brain Edema,Intracranial Pressure and Neurological Outcomes after Traumatic Brain Injury in Rat

Background:

In previous studies, the neuroprotective effect of 17β-estradiol in diffuse traumatic brain injury has been shown. This study used ICI 182,780, a non-selective estrogen receptor antagonist, to test the hypothesis that the neuroprotective effect of 17β-estradiol in traumatic brain injury is mediated by the estrogen receptors.

Methods:

The ovariectomized rats were divided into eight groups. Brain injury was induced by Marmarou’s method. Estrogen was injected 30 minutes after traumatic brain injury, and ICI 182,780 was injected before traumatic brain injury and also before estrogen treatment. In one group only ICI 182,780 was injected. The brain water content and Evans blue dye content were measured 24 and 5 hours after traumatic brain injury, respectively. The neurologic outcomes and intracranial pressure were assessed before, 4, and 24 hours after traumatic brain injury.

Results:

Brain water content and Evans blue content were less in estrogen-treated group comparison to vehicle group. ICI 182,780 eliminated the effects of estrogen on brain edema and brain blood barrier permeability. Intracranial pressure was increased significantly after trauma, and estrogen decreased intracranial pressure at 4 and 24 hours after traumatic brain injury in comparison to vehicle. This inhibitory effect was also eliminated by treatment with ICI182,780. ICI 182,780 also inhibited the estrogen induced increase in neurologic outcomes following traumatic brain injury. However, the use of ICI 182,780 alone had no neuroprotective effect after traumatic brain injury.

Conclusion:

The results suggest that classical estrogen receptors have probably a role in the neuroprotective function of estrogen following traumatic brain injury.Key Words: Estrogens, Intracranial pressure (ICP), Brain edema     

Germination and growth response of flax (Linum usitatissimum) to salinity stress by different salt types and concentrations

This investigation was conducted to explore the effects of salt types with different concentrations on germination and growth parameters of flax seeds. The experiment was set out as a factorial experiment based on a completely randomized design with three replications. We used six kinds of salts (NaCl, CaCl₂, CaCO₃, Na₂SO₄, Na₂CO₃ and KCl) with concentrations of 0, 50, 100 and 200 mM. According to the results, the inhibitory effects of the five salt types differed substantially, especially in the case of CaCO₃, Na₂CO₃ and Na₂SO₄. Inhibitory effects of these salts were very strong compared to those of NaCl and CaCl₂. Germination of flax seeds by various salts was in the order of NaCl > CaCl₂ > KCl > Na₂CO₃ > Na₂SO₄ > CaCO₃. The effect of salt concentration was obvious, too. Seeds of flax were able to germinate even in 200 mM NaCl, but they only germinated in distilled water or at very low CaCO₃, Na₂CO₃ and Na₂SO₄ concentrations (50 mM).