Effect of Bone Marrow Mesenchymal Stem Cell Sheets on Skin Capillary Parameters in a diabetic wound model: A Novel Preliminary Study

Background:Treatment with BMMSCs has anti-inflammatory, tissue regenerative, angiogenic, and immune-stimulating effects. When using as sheets or accumulate, BMMSCs causes the development of neoangiogenesis in damaged skin tissue. Diabetes, a metabolic disorder, can negatively affect many physiological functions, including the process of skin injury repair. This adverse impact may increase the risk of skin surgery. RSF is commonly used in reconstructive surgery. The terminal part of the RSF is often affected by necrosis because of impaired blood flow, which is exacerbated in diabetes. This study investigated the effect of stem cells, applied as accumulated or cell sheets, along with RSF surgery on skin capillaries in STZ-induced diabetic rats. Methods:Thirty male Wistar rats were divided into three groups (n = 10): diabetes-RSF control, diabetes-RSF local applied stem cells (loc-BMMSCs), diabetes-RSF applied stem cells as accumulated or cell sheets (ac-BMMSCs). Two weeks after the STZ injection, RSF surgery and stem cell therapy (6 × 10⁹) were carried out (day zero). Furthermore, stereological methods were used to investigate the capillary patterns among the groups. Anti-CD31/PCAM1 immunohistochemistry was also used for further confirmation of changes in capillary parameters. Results:The results demonstrated that capillaries were protected by MSC sheets in the flap tissue, and the thickness of the epidermal layer was improved, indicationg the possible beneficial effects of MSC sheets on diabetic wound treatment. Conclusion:Stem cells, as ac-BMMSCs, may decrease the levels of wound healing complications in diabetes and can be considered as a cell therapy option in such conditions.Key Words: Neoangiogenesis, Skin flap, Transplantation, Wound healing     

A new ENU-induced mutant mouse with defective spermatogenesis caused by a nonsense mutation of the syntaxin 2/epimorphin (Stx2/Epim) gene

Repro34 is an N-ethyl-N-nitrosourea (ENU)-induced mutation in mice showing male-specific infertility caused by defective spermatogenesis. In the present study, we investigated pathogenesis and molecular lesions in relation to spermatogenesis in the repro34/repro34 homozygous mouse. Histological examination of the testis showed that the seminiferous epithelium of the repro34/repro34 mouse contained spermatogonia and spermatocytes but no round and elongating spermatids. Instead of these haploid cells, multinucleated giant cells occupied the niche of the seminiferous tubules. Immunohistochemical staining for Hsc70t, an elongating spermatid specific protein, confirmed the absence of elongating spermatids. Furthermore, RT-PCR showed that there were significantly reduced expressions of the marker genes specifically expressed in the spermatid and that there was no difference in the expressions of the spermatocyte specific marker genes. These findings indicated interruption of the spermatogenesis during transition from the spermatocyte to spermatid in the repro34/repro34 mouse. The repro34 locus has been mapped on a 7.0-Mb region of mouse chromosome 5 containing the Syntaxin 2/Epimorphin (Stx2/Epim) gene, and targeted disruption of this gene has been reported to cause defective spermatogenesis. We therefore sequenced the entire coding region of the Stx2/Epim gene and found a nucleotide substitution that results in a nonsense mutation of this gene. The expression pattern of the Stx2/Epim gene during the first wave of spermatogenesis, increased expression at later stages of spermatogenesis, was in agreement with the affected phase of spermatogenesis in the adult repro34/repro34 testis. We therefore concluded that the male infertility of the repro34/repro34 mouse is caused by the interruption of spermatogenesis during transition from the spermatocyte to spermatid and that the nonsense mutation of the Stx2/Epim gene is responsible for the interruption of spermatogenesis.     

The ameliorating effects of polyamine supplement on physiological and biochemical parameters of Stevia rebaudiana Bertoni under cold stress

Stevia rebaudiana Bertoni is a perennial medicinal herb indigenous to tropical regions of South America. The present study evaluated some physiological changes of the plant under cold in the absence and the presence of polyamine conditions. The results showed photosynthetic pigments and photochemical efficiency of PSII reduced in cold treatment conditions than control, significantly. However, a considerable increase in the parameters was observed in all of polyamine treated goups. Leaf compatible solutes such as proline and glycine betaine that were not shown significantly increasing at cold treatment groups, elevated in the 2 days cold-polyamine-treated group. The similar results were seen for free amino acid and total protein contents of the treated plants. It was also revealed that specific activity of some antioxidant enzymes such as catalase and ascorbate peroxidase increased in cold-treated or cold + polyamine-treated groups than related controls. It caused a significant decrease in H₂O₂ and malondialdehyde contents of the treated plants. The results indicated an increasing in saturated long chain fatty acids after 4 days of cold at 4 °C. It was concluded that the polyamine supplement can induced a considerable tolerance in stevia plants at cold condition.     

Effects of fertilizers on plant diversity,density, and uniformity in Golestan rangelands

One of the management operations in rangeland ecosystems for pastures restoration and improvement is fertilizer application. This study was designed to investigate the effect of fertilizer (50 kg/ha urea and ammonium phosphate fertilizers) on species diversity changes in a 100-ha experimental field of Salmechal Rangeland, Golestan Province. Plant species mean canopy cover was recorded using a 50-m² sampling plot. Statistical analysis was performed using Excel 2007 and PAST software. Fertilizer application increased the number of species present in the region from 21 to 51 and the plant families from 11 to 23. The forbs had the highest proportion (54.91%) and bushes and shrubs had the lowest percentage in the region. Fertilizer application increased plant diversity, uniformity, and coverage of the grasses. There was a significant difference (p < 0.01) between the grass and forbs coverage before and after fertilizer treatments. The results showed that the biological and physiological growth periods of the annual and perennial plants were significantly different before and after fertilizer application (p < 0.01). Also, their proportion in rangeland plant species composition significantly increased after fertilizer application.     

Effect of chronic intracerebroventricluar administration of lipopolysaccharide on connexin43 protein expression in rat hippocampus

Background: Hippocampal damages, which are accompanied by inflammation, are among the main causes of epilepsy acquisition. We previously reported that chronic intracerebroventricular (i.c.v.) injection of lipopolysaccharide (LPS) modulates epileptogenesis in rats. There is a network of gap junction channels in the hippocampus that contribute to epileptogenesis. Gap junction channels are formed by oligomeric protein subunits called connexins (Cx). Astrocytic Cx43 and neuronal Cx36 are expressed in the hippocampus. In order to find out the possible role of gap junctions in seizure-modulating effect of LPS and neuroinflammation, we studied the effect of central administration of LPS on expression of Cx36 and Cx43 in rat hippocampus. Methods: LPS, 2.5 µg/rat/day, was injected i.c.v. to male Wistar rats for 14 days. mRNA and protein abundance of Cx36, Cx43 and IL1-β were measured in rat hippocampus by real time-PCR, Western blot and ELISA techniques, at the beginning, in the middle, and at the end of the treatment period. Results: IL1-β protein level was significantly increased 6 h after first injection of LPS. Cx36 and Cx43 mRNA expression did not alter during chronic administration of LPS. A selective decrease in Cx43 protein expression was observed after 7 injections of LPS. Conclusion: It is suggested that Cx43 containing gap junctions in the hippocampus is down-regulated in response to chronic injection of LPS. This event can inhibit propagation of toxic and noxious molecules to neighboring cells and modulate hippocampal excitability and epileptogenesis. Key Words: Connexin36 (Cx36), Connexin43 (Cx43), Interleukin-1β (IL1-β )     

Electrodeposition of homogeneous and adherent polypyrrole/Na<Superscript>+</Superscript>-cloisite nanocomposite on iron electrodes

In this work homogeneous and adherent nanocomposite of polypyrrole (PPY) with Na⁺-Cloisite nanoclays was successfully prepared by insitu electropolymerization of pyrrole monomer in the presence of Na⁺-Cloisite nanoclays. Formation of nanocomposite and incorporation of nanoclays in polypyrrole matrix was endorsed and characterized by FTIR spectroscopy studies, X-ray diffraction (XRD) pattern and scanning electron microscopy (SEM). The X-ray difraction patterns showed that PPY was intercalated between clay layers in the order of nanoscale. The morphology of the resulted nanocomposite was studied by scanning electron micrograph (SEM). Cyclic voltammetry experiments were carried out in different electrolytes and showed that nanocomposite is electroactive, while conductivity of the nanocomposite derived from four point probe technique indicated that the resulted nanocomposite is conductive. The anticorrosive properties of a thin layer coating of PPY/Na⁺-Cloisite nanocomposite on iron coupons was evaluated and compared with pure polypyrrole coating. According to the results in various corrosive environments PPY/Na⁺-Cloisite nanocomposite has enhanced corrosion protection effect in comparison to pure polypyrrole coating. It was observed that the corrosion protection effect was due to the dispersion of nanoclays in polymer matrix, which results in the increase in the corrosion potential and decrease in corrosion current and corrosion rate.     

Use of Selection Indices Based on Multivariate Analysis for Improving Grain Yield in Rice

In order to study selection indices for improving rice grain yield, a cross was made between an Iranian traditional rice (Oryza sativa L.) variety, Tarommahalli and an improved indica rice variety, Khazar in 2006. The traits of the parents (30 plants), F1 (30 plants) and F2 generations (492 individuals) were evaluated at the Rice Research institute of Iran (RRII) during 2007. Heritabilities of the number of panicles per plant, plant height, days to heading and panicle exsertion were greater than that of grain yield. The selection indices were developed using the results of multivariate analysis. To evaluate selection strategies to maximize grain yield, 14 selection indices were calculated based on two methods (optimum and base) and combinations of 12 traits with various economic weights. Results of selection indices showed that selection for grain weight, number of panicles per plant and panicle length by using their phenotypic and/or genotypic direct effects (path coefficient) as economic weights should serve as an effective selection criterion for using either the optimum or base index.     

Ileal relaxation induced by Mentha longifolia (L.) leaf extract in rat

The effect of Mentha longifolia (L.) leaf hydroalcoholic extract (MLE) was examined on rat ileal smooth muscle contractions. Last portion of ileum from male adult Wistar rat was mounted in an organ bath containing Tyrode solution. The tissue was contracted by carbachol (CCh, 10 microM), KCl (60 mM) and BaC12 (4 mM) and then MLE (0.0625-1 mg mL(-1)) was added to the bath cumulatively. The effect of MLE on KCl-induced contraction was examined after tissue incubation with propranolol (1 microM), naloxone (1 microM) and N omega-nitro-L-arginine methyl ester (L-NAME, 100 microM). The effect of MLE on CaCl2-induced ileal contraction in Ca(2+)-free with high potassium Tyrode solution was also evaluated. The role of potassium channels was examined by ileum incubation (5 mim) with tetraethylammonium (TEA, 1 mM). The results showed that KCl-, CCh and BaCl2-induced ileal contractions were inhibited (p < 0.001) by cumulative concentrations of MLE with the same potency. In addition, MLE (0.25-1 mg mL(-1)) inhibited (p < 0.01) ileal contractions induced by CaCl2 (0.45-2.7 mM) in a concentration-related manner. The antispasmodic effect of MLE was affected neither by propranolol, L-NAME nor by naloxone. The MLE concentration-response curve was shifted to the right (p < 0.05) by tissue incubation with TEA. From results it may be suggested that Mentha longifolia hydroalcoholic leaf extract induces its spasmolytic activity mainly through disturbance in calcium mobilization and partly by potassium channels activation. Present results show that Mentha longifolia leaf extract exerts relaxant effects on intestinal smooth muscle, consistent with the traditional use of the plant to treat gastrointestinal disorders such as diarrhea and colic.     

Interaction of phosphodiesterase 5 inhibitor with malathion on rat brain mitochondrial-bound hexokinase activity

The main objective of this study was to investigate the possible protective effect of pentoxifylline as a phosphodiesterase 5 inhibitor used as a cardiovascular medication on malathion-induced changes on rat mitochondrial-bound hexokinase activity.Animals in four various groups received moderate toxic dose of malathion (200 mg/kg/day), effective dose of pentoxifylline (50 mg/kg/day) alone and in combination, and the control group that received only vehicle. All administrations were done intraperitoneally for one week. At the end of the experiment, the brain was removed and the mitochondria were isolated. Hexokinase (HK) activity, cellular lipid peroxidation (LPO) and total antioxidant capacity (TAC) were analyzed in brain mitochondria.Malathion noticeably decreased TAC and increased HK activity and LPO in the mitochondria whereas pentoxifylline significantly restored malathion-induced changes in LPO, HK, and TAC.The results of the present study indicate that phosphodiesterase 5 inhibition remarkably protects brain mitochondria from malathion-induced changes on HK activity and oxidative stress.     

Effects of Artemisia annua L. (Asteracea) on nutritional physiology and enzyme activities of elm leaf beetle, Xanthogaleruca luteola Mull. (Coleoptera: Chrysomellidae)

Methanolic extract of Artemisia annua L., a weed collected around paddy fields in north of Iran, was investigated for its toxic effects on: feeding, growth, fecundity, fertility including the biochemical characteristics of elm leaf beetle Xanthogaleruca luteola Mull. The toxicity tests indicate that insecticidal effect was 2.4-fold higher in 3rd instar larvae than in adults. The LC₅₀ was 48% and 43.77% of a methanolic leaf extract at 24 and 48 h, respectively. The feeding deterrence was highest at 10% concentration and lowest at 0.625% for 3rd instar larvae and adults during the first 24 h and decreased thereafter. Larval duration significantly increased at higher concentrations. Larval treatment with 10% methanolic extract resulted in deformed adults (adultoids) with drastic changes in gross morphology of female reproductive system. Food digestion and assimilation significantly decreased in adults. Larval duration and adult fecundity and fertility were also significantly reduced. Twenty-four hours after treating 3rd instar larva with the extract the levels of glucose, protein, urea, uric acid, α-amylase, alkaline phosphatase, alanine amino transferase and aspartate amino transferase significantly changed. However, at 48 h the extract lost its potency. The extract did not effect the level of cholesterol in treated larva. These results indicate that Artemisia annua L. extract contains inhibitors of key metabolic pathways that may be useful in future control of the elm leaf beetle.