Electrospun triple-layered PLLA/gelatin. PRGF/PLLA scaffold induces fibroblast migration

The function of fibroblast cells in wounded areas results in reconstruction of the extra cellular matrix and consequently resolution of granulation tissue. It is suggested that the use of platelet-rich plasma can accelerate the healing process in nonhealing or slow-healing wounds. In this study, a simple and novel method has been used to fabricate an electrospun three-layered scaffold containing plasma rich in growth factor with the aim of increasing the proliferation and migration of fibroblast cells in vitro. First, plasma rich in growth factor was derived from platelet rich plasma, and then a three-layered scaffold was fabricated using PLLA nanofibers as the outer layers and plasma rich in growth factor-containing gelatin fibers as the internal layer. The growth morphology of cells seeded on this scaffold was compared to those seeded on one layered PLLA scaffold. The study of the cell growth rate on different substrates and the migration of cells in response to the drug release of multilayered scaffold was investigated by the cell quantification assay and a modified under agarose assay. Scanning electron microscopy and fluorescence images showed that cells seeded on multilayered scaffold were completely oriented 72 hours after seeding compared to those seeded on PLLA scaffold. The cell quantification assay also indicated significant increase in proliferation rate of cells seeded on three-layered scaffold compared to those seeded on PLLA scaffold and finally, monitoring cell migration proved that cells migrate significantly toward the three-layered scaffold up to 48 to 72 hours and afterwards start to show a diminished migration rate toward this scaffold.     

The effects of preservation procedures on amniotic membrane's ability to serve as a substrate for cultivation of endothelial cells

Amniotic membrane (AM) has been used as a scaffold for the ex vivo expansion of different types of cells and a cell delivery matrix in regenerative medicine. Since the preservation procedures can influence the AM properties for experimental and clinical purposes, this study was established to investigate the feasibility of using the AM after different preservation methods to serve as substrates for endothelial cell expansion ex vivo. The effects of cryopreservation and lyophilization were evaluated on mechanical and histological characteristics of the AM, and the results were compared with the fresh AM. The ECM components of the basement membrane were well conserved in all groups. Although lyophilization resulted in more histological changes and lower level of physical variables including thickness, F_max, elongation at break and suture retention than the fresh and cryopreserved AM, endothelial cells grown on the lyophilized AM were better attached to the basement membrane. Cytotoxicity assay by MTT showed that the lyophilized AM is a compatible substrate for endothelial cells cultivation. The findings of this study suggest that the lyophilized AM is a suitable matrix for cultivation of endothelial cells due to this fact that lyophilization led to exposure of basement membrane of the AM.     

Lithium reverses the effect of opioids on eNOS/nitric oxide pathway in human umbilical vein endothelial cells

The main challenge of pain management with opioids is development of acute and chronic analgesic tolerance. Several studies on neuronal cells have focused on the molecular mechanisms involved in tolerance such as cyclic AMP (cAMP) activation, and nitric oxide (NO) pathway. However, the effects of opioids on non-neuronal cells and tolerance development have been poorly investigated. Lithium chloride is a glycogen synthase kinase 3β (GSK-3β) inhibitor and exert its effects through modulation of nitric oxide pathway. In this study we examined the effect of lithium on acute/chronic morphine and methadone administration in endothelial cells which express mu opioid receptors. Human umbilical vein endothelial cells (HUVECs) were treated with different doses of morphine, methadone, and lithium for six and 48 h. Then we evaluated cell viability, nitrite and cyclic AMP levels, as well as the expression of endothelial nitric oxide synthase (eNOS) protein using Immunocytochemistry (ICC) assay and phosphorylated GSK-3β enzyme by western blot analysis in cells. Both chronic morphine and methadone treatment increased NO level and eNOS expression in HUVECs. Morphine induced cAMP overproduction after 48 h exposure with cells. Lithium pretreatment (10 mM) in both morphine and methadone received groups significantly reduced nitrite and cAMP levels as well as eNOS expression as compared to the control. The decreased amount of phospho GSK-3β due to the opioid exposure was increased following lithium treatment. Tolerance like pattern may occur in non-neuronal cells with opioid receptors and this study clearly revealed the attenuation of morphine and methadone tolerance like behavior by lithium treatment in HUVECs.

     

Overexpression of reactive oxygen species modulator 1 is associated with advanced grades of bladder cancer

Molecular Biology Reports - Reactive Oxygen Species Modulator 1 (ROMO1) plays a pivotal role in the regulation of mitochondrial structure integrity, and the production of reactive oxygen species...     

Association of the colorectal cancer and <Emphasis Type="Italic">MDR1</Emphasis> gene polymorphism in an Iranian population

The human multidrug resistance (MDR1) gene product P-glycoprotein is highly expressed in intestinal epithelial cells, where it constitutes a barrier against xenobiotics, bacterial toxins, drugs and other biologically active compounds, possibly carcinogens. In this study, an association of MDR1 gene polymorphism and the occurrence of colorectal cancer were evaluated. In this case-control-designed 118 unrelated colorectal cancer and 137 sex-and-ages matched healthy controls were enrolled. The C3435T MDR1 gene polymorphism was identified using the polymerase chain reaction-restriction fragment length polymorphism method. Significantly increased frequencies of the 3435T allele and the 3435TT were observed in patients with colorectal cancer compared with controls (P = 0.03; OR, 95% CI; 1.46 for 3435T allele and P = 0.003; OR, 95% CI; 2.2 for 3435TT genotype). In contrast, frequency of genotype TT was significantly higher in controls compared to colorectal cancer (P = 0.006; OR, 95% CI; 0.49 for TC genotype). In this study suggest that C3435T MDR1 polymorphism has an association with colorectal cancer. The results support that the presence of allele C results in decreased susceptibility to colorectal cancer.     

Length‐weight relationships of Turcinoemacheilus kosswigi Bănărescu and Nalbant, 1964 (Teleostei:Nemacheilidae) in southwestern Iran and its relation to habitat structure

Length‐weight relationships of Turcinoemacheilus kosswigi, based on 56 individuals from two newly discovered localities in the Iranian Tigris drainage, are reported for the first time. These new localities have the same structural characters as already‐reported habitats, but the species was found in much higher altitudes than before.     

A study on common bean cultivars to identify sources of resistance against the black bean aphid,Aphis fabae Scopoli (Hemiptera: Aphididae)

Black bean aphid is an important common bean pest in the world. Aphids damage crops both directly by their feeding and by spreading viruses. Also, they indirectly damages with excretion honeydew and smokes moulds growth through some physiological processes and transmission of viruses. Resistant cultivars application is the main strategy to control Aphis fabae Scopoli. In this experiment, different lines and cultivars were infested with five wingless adult aphids. After 7 and 14?days, plants were evaluated and aphid population was counted and recorded. After 45?days when plants reached to the flowering stage, test was repeated. Result showed that there was a significant difference among cultivars in number of adult aphid in two-leaf stage. Most of the number of adult at the first week in two-leaf stage observed on Goynok cultivar that had significant difference to other lines and cultivars. Number of adults and nymphs among the line and cultivar at flowering stage did not show significant difference. Number of deployed aphids after two weeks was significantly higher than the first week that observed in all cultivars. Totally, the most resistant cultivar was Sayad and the most susceptible cultivar was Goynok.     

CFTR haplotypes in northern Iranian population

Background

Cystic fibrosis (CF) is a multiorganic autosomal recessive disorder, caused by mutation in cystic fibrosis transmembrane conductance regulator (CFTR). CF is highly heterogeneous in Iranian population and molecular diagnosis based on direct identification of mutations is not completely efficient. The use of polymorphic intragenic markers not only can facilitate phenotype prediction in prenatal diagnosis by gene tracking, but also can lead to the demonstration of possible associations between haplotypes and specific mutations.

Methods

60 CF patients and 53 fertile normal subjects originating from North of Iran were analyzed for F508del mutation and c.1210-12T(5_9), c.1408A>G and c.744-33GATT(6_8) polymorphisms.

Results

c.1210-12T[7] is the most prevalent allele in normal individuals and CF non-F508del patients with 87.7%and 86.7% frequencies respectively. c.1408A>G survey showed that frequency of allele G and A is nearly equal in both non-F508del CF patients and normal individuals. c.744-33GATT(6_8) study showed that 7 repeat is the most prevalent allele in normal individuals and non-F508del CF patients with 80.2% and 82.1% frequencies respectively. The [c.1408A; c.1210-12T[9]; c.744-33GATT[6]] haplotype was only associated with mutant alleles including F508del.

Conclusions

The allelic distribution and heterozygosity results suggest that c.1408A>G, c.1210-12T(5_9) and c.744-33GATT(6_8) can contribute to carrier detection and prenatal diagnosis of CF in Iranian families with previous history of the disease.     

Effects of Simple and Microencapsulated <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> With or Without Inulin on the Broiler Meat Quality Infected by Avian Influenza Virus (H<Subscript>9</Subscript>N<Subscript>2</Subscript>)

The aims of the present study were to investigate the effects of simple and microencapsulated Lactobacillus acidophilus as probiotic with or without inulin as prebiotic on meat quality of broiler infected by avian influenza virus (H₉N₂). Two hundred-day-old chicks were randomly allocated into 14 groups based on simple, microencapsulated probiotic and prebiotic and based on vaccination and challenge with H₉N₂ virus. Groups 1–7 contained 20 chicks, and groups 8–12 and 14 contained 10 chicks. Group 13 was derived from group 1 with 10 chicks at challenge day with avian influenza virus (AIV). Half of the groups were vaccinated by H₉N₂ vaccine on day 5. All groups except the negative control and positive vaccine control were challenged with 10^6.5 EID₅₀ of low-virulence H₉N₂ AIV at day 21. Each bird was received 10⁹ CFU of simple or microencapsulated probiotic on days 0 and 17 by gavage. Prebiotic as dose as 0.1 % of feed weight was used daily. Increase in water-holding capacity, dry matter, ash and protein content, and decrease in dripping loss plus beneficial changes in lightness and redness of breast meat were detected in response to probiotic especially microencapsulated synbiotic. In conclusion, probiotic alone or with prebiotic was able to improve the physicochemical properties of chicken breast muscle in both healthy and AIV-infected chickens.     

Determination of human albumin in serum and urine samples by constant‐energy synchronous fluorescence method

A sensitive spectrofluorimetric method using constant‐energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1–220.0 µg mL^–1 of albumin with a detection limit of 7.0 × 10^–3 µg mL^–1. The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL^–1 albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples. Copyright © 2014 John Wiley & Sons, Ltd.