Pelagia noctiluca (Scyphozoa) Crude Venom Injection Elicits Oxidative Stress and Inflammatory Response in Rats

Cnidarian toxins represent a rich source of biologically active compounds. Since they may act via oxidative stress events, the aim of the present study was to verify whether crude venom, extracted from the jellyfish Pelagia noctiluca, elicits inflammation and oxidative stress processes, known to be mediated by Reactive Oxygen Species (ROS) production, in rats. In a first set of experiments, the animals were injected with crude venom (at three different doses 6, 30 and 60 µg/kg, suspended in saline solution, i.v.) to test the mortality and possible blood pressure changes. In a second set of experiments, to confirm that Pelagia noctiluca crude venom enhances ROS formation and may contribute to the pathophysiology of inflammation, crude venom-injected animals (30 µg/kg) were also treated with tempol, a powerful antioxidant (100 mg/kg i.p., 30 and 60 min after crude venom). Administration of tempol after crude venom challenge, caused a significant reduction of each parameter related to inflammation. The potential effect of Pelagia noctiluca crude venom in the systemic inflammation process has been here demonstrated, adding novel information about its biological activity.     

4H12, a Murine Monoclonal Antibody Directed against Myosin Heavy Chain-9 Expressed on Acinar Cell Carcinoma of Pancreas with Potential Therapeutic Application

Background:PACC is a rare type of pancreatic exocrine neoplasm that is frequently diagnosed at late stages with a high rate of metastasis. Identification of new biomarkers for PACC can improve our knowledge of its biology, early detection, or targeted therapy. In this study, hybridoma technology was used to generate mAbs against Faraz-ICR, a pancreatic acinar cell carcinoma cell line. Methods: Cell ELISA and flow cytometry were used for screening, and the 4H12 hybridoma clone was selected for further analysis. The 4H12 mAb was specific for MYH9 as determined by Immunoprecipitation, Western blot, and mass spectrometry. Results:This antibody reacted variably with other cancer cells, in comparison to Faraz-ICR cell. Besides, by immunohistochemical staining, the acinar cell tumor, which was the source of Faraz-ICR, showed high MYH9 expression. Among 21 PDAC cases, nine (42.8%) expressed MYH9 with low intensity, while 10 (47.8%) and 2 (9.5%) cases expressed MYH9 with moderate to strong intensities, respectively. The 4H12 mAb inhibited the proliferation of Faraz-ICR cells in a dose-dependent manner from 0.75 to 12.5 μg/ml concentrations (p < 0.0001 and p < 0.002). IC₅₀ values were achieved at 12.09 ± 4.19 µg/ml and 7.74 ± 4.28 µg/ml after 24- and 48-h treatment, respectively. Conclusion:Our data suggest that the 4H12 mAb can serve as a tool for investigating the role of MYH9 pancreatic cancer biology and prognosis. Key Words: Acinar cell carcinoma, Biomarkers, Monoclonal antibody, Pancreas     

Impact of Methylenetetrahydrofolate Reductase C677T Polymorphism on the Risk of Gastric Cancer and Its Interaction with Helicobacter pylori Infection

Background

Attempts for early detection of gastric cancer have recently focused on host''s genetic susceptibility factors and gene-environment interactions. We have, herein, studied the association of MTHFR C677T single nucleotide polymorphism (SNP) and its interaction with Helicobacter pylori infection, smoking, age and gender on the risk of gastric cancer among an Iranian population.

Methods

Gastric cancer patients (n = 450) and cancer-free controls (n = 780) were studied for serum H. pylori-specific IgG antibodies by ELISA and MTHFR C677T polymorphism (SNP) by PCR-RFLP. Demographic and life style data were collected through patient interviews. Unconditional logistic regression model estimated odds ratio (OR) and the corresponding 95% confidence intervals (CI).

Results

The interactions of MTHFR genotype with H. pylori infection (P = 0.03), age (P = 0.049) and gender (P = 0.007) were statistically significant. Accordingly, MTHFR C677T carriers who were also positive for H. pylori infection exhibited 80% (OR = 1.8, 95% CI = 1.0-2.9) significant excess risk of non-cardia gastric cancer. Furthermore, subjects over the age of 50 or female subjects carrying MTHFR C677T SNP showed 40 (OR = 1.4, 95% CI = 1.0-2.0) and 100 (OR = 2.0, 95% CI = 1.2-3.2) percent increased risk of gastric cancer, respectively.

Conclusion

MTHFR C677T SNP seems to increase the risk of gastric cancer and the effect is significantly inflated by interactions with H. pylori infection, age and gender. Key Words: Helicobacter pylori, Smoking, Gender identity, Age group, Methylenetetrahydrofolate reductase     

Production performance,immune response and blood biochemical parameters in broiler chickens fed diet incorporated with prebiotics

This experiment was conducted to identify the suitable replacer of broiler feed antibiotics with prebiotics (mannan oligosaccharides‐MOS or fructooligosaccharide‐FOS). Two hundred and forty (240)‐day‐old chicks were randomly divided into 30 groups (6 treatments x 5 replicates/treatment x 8 chicks/replicate). Six experimental diets T₁, T₂, T₃, T₄, T₅ and T₆ were formulated to contain an additional 0, antibiotic, that is, bacitracin methylene di‐salicylate (BMD) @20 mg/kg, MOS (0.1% and 0.2%) and FOS (0.1% and 0.2%) respectively. Body weight gain was significantly (p < 0.05) increased in MOS‐0.2% supplemented group at 0–21 d and 0–42 d of broiler chicken. Humoral and in vivo cell‐mediated immune response were significantly improved (p < 0.05) in BMD, MOS @0.1% or 0.2% treated group. Significant (p < 0.05) increase was recorded in total protein (except 21 d), albumin and aspartate amino transferase (AST) and decrease (p < 0.05) in alanine amino transferase (except 42 d), cholesterol and uric acid concentration. The weight of breast, thigh, back, drumstick bursa of Fabricius and thymus were higher (p < 0.05) in the birds given the MOS @0.2% (T₄). It is concluded that MOS @0.2% may be suitable replacer of antibiotic growth promoter, and it has a beneficial effect on production performance, immune responses, blood biochemical parameters and cut up parts in broiler chickens.     

Dyeing behavior of polyacrylonitrile nanofibers: Physicochemical parameters of dyeing with basic dye in comparison with regular polyacrylonitrile fibers

Polyacrylonitrile nanofibers were produced using the electrospinning method and dyed with a basic dye alongside regular polyacrylonitrile fibers. In order to investigate the effect of high surface area to volume ratio of nanofibers on their adsorption behavior in comparison with regular fibers, the dyeing conditions for both types of fibers were kept just the same. Physiochemical parameters of dyeing such as adsorption isotherm, standard affinity, enthalpy change, rate of dyeing constant, diffusion coefficient, and activation energy of diffusion were investigated for both types of fibers. The results showed that the adsorption process can be well described with the Langmuir adsorption isotherm for both types of fibers whereas the standard affinity of dye to nanofibers was higher than regular fibers and the higher negative values of enthalpy changes were obtained for regular fibers. The nanofibers rate of dyeing was faster than regular fibers with higher amounts of diffusion coefficients and lower amounts of activation energy of diffusion. This study also revealed that in spite of the approximately same amount of dye exhaustion for both types of fibers, the color strength of regular fibers was noticeably higher than nanofibers.     

Antimicrobial Susceptibility of 41 Burkholderia mallei Isolates From Spontaneous Outbreaks of Equine Glanders in Punjab,Pakistan

Minimum inhibitory concentrations (MICs) of 20 antimicrobial agents for 41 isolates of Burkholderia mallei from natural outbreaks of equine glanders were determined by agar dilution. All isolates were intrinsically resistant to ampicillin (MIC₉₀ ≥128). Resistance to other antimicrobials was as follows: 95.1% to amoxicillin and cephradine, 85.4% to cefuroxime and norfloxacin, 68.3% to ceftizoxime and ceftriaxone, 61.1% to ceftiofur, 58.5% to oxytetracycline, 41.5% to ciprofloxacin, 58.5% to roxithromycin, 17.1% cefotaxime, and 12.2% clarithromycin. Overall resistance patterns revealed that 17% of isolates were simultaneously resistant to amoxicillin, cephradine, cefuroxime, ceftizoxime, ceftriaxone and norfloxacin. None of the isolates were resistant to amoxicillin-clavulanic acid, doxycycline, chloramphenicol, gentamicin or trimethoprim-sulphadiazine. Mode MICs for these antimicrobials were 2, 1, 8, 4 and 1 μg/ml, respectively. A majority of the isolates (∼ 94%) were susceptible to both enrofloxacin and ofloxacin. These data provide an updated perspective on susceptibility profiles of current strains of B. mallei in an endemic area.     

Ursene type nortriterpene from Debregeasia salicifolia

A new triterpene, 3beta-19alpha-dihydroxy-30-norurs-12-ene (1), has been isolated from the methanolic extract of Debregeasia salicifolia stems. Its structure has been elucidated with the help of one- and two-dimensional NMR and NOE studies. Lupeol, beta-sitosterol, stigmasterol and oleanolic acid are also reported for the first time from this species.     

Morphometrical study of polysialylated neural cell adhesion molecule positive cells in rat pups hippocampus following induction of seizure during pregnancy

Background:The polysialylated neural cell adhesion molecule (PSA-NCAM) is expressed in developing brain. Fetal brain damage is caused by different conditions such as seizure and hypoxia. The present study was designed to investigate the effect of maternal seizures on the number of PSA-NCAM positive cells in pup''s hippocampus. Methods: Female Wistar rats were divided into four groups: (a) kindled rats which received PTZ (40 mg/kg, i.p.) during pregnancy from embryonic day 14-19 (E14-E19) every 48 h, (b) kindled rats which did not receive PTZ during pregnancy, (c) non-kindle, pregnant rats which received PTZ injection (40 mg/kg, i.p.) during pregnancy from E14 to E19 every 48 h, and (d) non-kindle, pregnant rats which received injection with an equal volume of normal saline as sham controls. At postnatal day 14 (PD₁₄), rat pups were perfused, and their brain were fixed, embedded and coronal sections stained by immunohistochemistry method. The number of PSA-NCAM positive cells per unit area in the pup''s hippocampus was counted. Results: The number of PSA-NCAM positive cells in the CA1, CA3, and DG fields of pup''s hippocampus, which was obtained from mothers who experienced PTZ injection during pregnancy, was decreased approximately 2.6 (P = 0.001), 2 (P = 0.001), and 2.1 (P = 0.001) times compared with non-PTZ treated maternal groups, respectively. Conclusions: Our study showed that maternal seizures reduced the number of neurons and also PSA-NCAM positive cells per unit area in the offspring hippocampus that it may cause impairment in hippocampal functions.Key Words: Epilepsy, Polysialylated neural cell adhesion molecule (PSA-NCAM), Hippocampus, Seizures     

Human Granulocyte Colony-Stimulating Factor (hG-CSF) Expression in Plastids of Lactuca sativa

Background: Human granulocyte colony-stimulating factor (hG-CSF) can serve as valuable biopharmaceutical for research and treatment of the human blood cancer. Transplastomic plants have been emerged as a new and high potential candidate for production of recombinant biopharmaceutical proteins in comparison with transgenic plants due to extremely high level expression, biosafety and many other advantages. Methods: hG-CSF gene was cloned into pCL vector between prrn16S promoter and TpsbA terminator. The recombinant vector was coated on nanogold particles and transformed to lettuce chloroplasts through biolistic method. Callogenesis and regeneration of cotyledonary explants were obtained by Murashige and Skoog media containing 6-benzylaminopurine and 1-naphthaleneacetic acid hormones. The presence of hG-CSF gene in plastome was studied with four specific PCR primers and expression by Western immunoblotting. Results: hG-CSF gene cloning was confirmed by digestion and sequencing. Transplastomic lettuce lines were regenerated and subjected to molecular analysis. The presence of hG-CSF in plastome was confirmed by PCR using specific primers designed from the plastid genome. Western immunoblotting of extracted protein from transplastomic plants showed a 20-kDa band, which verified the expression of recombinant protein in lettuce chloroplasts. Conclusions: This study is the first report that successfully express hG-CSF gene in lettuce chloroplast. The lettuce plastome can provide a cheap and safe expression platform for producing valuable biopharmaceuticals for research and treatment.Key Words: Plastid, Human granulocyte colony-stimulating factor (hG-CSF), Biolistics, Gene targeting