ABSTRACT
Background: vascular endothelial growth factor [VEGF] plays an important role in development of new blood vessel and angiogenesis. Human VEGF121is smallest member of VEGF family. Production of active and correct form of VEGF is the m6st challenging issue
Methods: here we described a method for optimization of refolding of VEGF121 which was expressed in bacterial host. Enzyme linked immunosorbent assay and proliferation assay of human endothelial cells was performed to monitor refolding and functional assay ofVEGFlz1
Results: using described method VEGF was in correct fold and detected by antibody in ELISA. Furthermore, VEGF stimulated proliferation of human endothelial cells in dose-dependent manner
Conclusion: refolded VEGF has potential for stem cell differentiation
ABSTRACT
<p><b>OBJECTIVE</b>Scorpion (Hemiscorpius lepturus) stings are a public health concern in Iran, particularly in south and southwestern regions of Iran. The gold standard for the treatment of a scorpion sting is anti-venom therapy. However, immunotherapy can have serious side effects, such as anaphylactic shock (which can sometimes even lead to death). The aim of the current study was to demonstrate the protective effect of ozone against toxicity induced by Hemiscorpius lepturus (H. lepturus) venom in mice.</p><p><b>METHODS</b>Eight hours after the injection of ozone to the experimental design groups, the male mice were decapitated and mitochondria were isolated from five different tissues (liver, kidney, heart, brain, and spinal cord) using differential ultracentrifugation. Then, assessment of mitochondrial parameters including mitochondrial reactive oxidative species (ROS) production, mitochondrial membrane potential (MMP), ATP level, and the release of cytochrome c from the mitochondria was performed.</p><p><b>RESULTS</b>Our results showed that H. lepturus venom-induced oxidative stress is related to ROS production and MMP collapse, which is correlated with cytochrome c release and ATP depletion, indicating the predisposition to the cell death signaling.</p><p><b>CONCLUSION</b>In general, ozone therapy in moderate dose can be considered as clinically effective for the treatment of H. lepturus sting as a protective and antioxidant agent.</p>
Subject(s)
Animals , Male , Mice , Brain , Metabolism , Cytochromes c , Metabolism , Heart , Kidney , Metabolism , Liver , Metabolism , Membrane Potential, Mitochondrial , Mice, Inbred BALB C , Muscle, Skeletal , Metabolism , Myocardium , Metabolism , Ozone , Pharmacology , Scorpion Venoms , Toxicity , Scorpions , Physiology , Spinal Cord , MetabolismABSTRACT
OBJECTIVE@#To express human vascular endothelial growth factor121 (VEGF121) in insect cells.@*METHODS@#A gene construct containing VEGF was cloned in the pFastBac-HTA vector, followed by transformation in DH10BAC. The recombinant bacmid was then extracted, and transfected into Sf9 insect cells. The transfected cells were harvested, and then VEGF expression was confirmed by western blotting using specific antibodies. The tube formation assay was used for functional assessment of VEGF.@*RESULTS@#Our results showed that VEGF could be successfully expressed in the baculovirus system. Purified VEGF was able to stimulate in vitro tube formation of human endothelial cells.@*CONCLUSIONS@#Results from this study demonstrated that the recombinantly-produced VEGF can be considered as a promising candidate for therapeutic purposes.
ABSTRACT
Objective To express human vascular endothelial growth factor121 (VEGF121) in insect cells. Methods A gene construct containing VEGF was cloned in the pFastBac-HTA vector, followed by transformation in DH10BAC. The recombinant bacmid was then extracted, and transfected into Sf9 insect cells. The transfected cells were harvested, and then VEGF expression was confirmed by western blotting using specific antibodies. The tube formation assay was used for functional assessment of VEGF. Results Our results showed that VEGF could be successfully expressed in the baculovirus system. Purified VEGF was able to stimulate in vitro tube formation of human endothelial cells. Conclusions Results from this study demonstrated that the recombinantly-produced VEGF can be considered as a promising candidate for therapeutic purposes.
ABSTRACT
Background: Baculovirus expression system is one of the most attractive and powerful eukaryotic expression systems for the production of recombinant proteins. The presence of a biomarker is required to monitor transfection efficiency or protein expression levels in insect cells
Methods: The aim of this study was to construct a baculovirus expression vector encoding a copepod super green fluorescent protein [copGFP]. In this light, the resultant vector was constructed and used for transfection of Spodoptera frugiperda cells
Results: Expression of the copGFP protein in insect cells was confirmed by fluorescent microscopy and Western-blot analysis
Conclusion: The application of copGFP control bacmid can be considered as an appropriate control for insect cell transfection
ABSTRACT
Background: During the recent years, significant progress has been achieved on development of novel anti-viral drugs. Natural products are assumed as the potential sources of novel anti-viral drugs; therefore, there are some previous studies reporting the anti-viral compounds from venomous animals. Based on the significant value for tracing of non-toxic anti-viral agents from natural resources, this study was aimed to investigate the anti-viral activity of some HPLC purified fractions derived from the venom of Iranian scorpion, Hemiscorpius lepturus, against human immunodeficiency virus 1 [HIV-1] and herpes simplex virus 1 [HSV-1]
Methods: H. Lepturus crude venom was subjected to reverse phase HPLC analysis to determine its active components precisely where four dominant fractions obtained at retention time of 156-160 minutes. The phospholipase A2 and hemolytic activities of the purified fractions were first evaluated. Then the anti-viral activity was measured using single cycle HIV [NL4-3] replication and HSV [KOS] plaque reduction assays
Results: The H. lepturus crude venom inhibited HIV replication by 73% at the concentration of 200 micro g/ml, while it did not show significant anti-HSV activity. It also inhibited the cell-free viral particles in a virucidal assay, while it showed no toxicity for the target cells in a proliferation assay. The four HPLC fractions purified from H. lepturus inhibited HIV with IC50 of 20 micro g/ml
Conclusion: H. lepturus venom contains components with considerable anti-HIV activity insofar as it has virucidal activity that offers a novel therapeutic approach against HIV infection. Our results suggest a promising pilot for anti-HIV drug discovery with H. lepturus scorpion venom
ABSTRACT
OBJECTIVE@#To evaluate a) heart place and tail length, b) their correlations with other biometrics, c) sexual differences in those features, in regard to cardiovascular system in a number of snakes from Iran, about which there is little information.@*METHODS@#We studied the fresh mortalities of snakes including 14 Naja oxiana (N. oxiana), 23 Macrovipera lebetina, and one male Montivipera latifii acquired from the Serpentatium of Pasteur Institute of Iran. In this respect, each specimen first was weighted, and then its ventral side of the integument was incised, and heart place was measured. Subsequently, other biometrical features such as total length, TAL, and snout-vent length were measured.@*RESULTS@#The results showed that heart place in N. oxiana, Macrovipera lebetina and Montivipera latifii was about 18%, 32% and 30%, and also TAL constituted about 16%, 11% and 7% of total body length, respectively. Moreover, females indicated anterior heart place and shorter tail than males. Furthermore, the measures and correlations indicated few differences between N. oxiana and typical terrestrial species.@*CONCLUSIONS@#The results denoted that in order to overcome hemocirculatory perturbations in vertical orientation while hooding and head raising behavior, N. oxiana need to have more important features than short heart to head distance and long tail. In addition, it gave the sexual differences in heart place and tail length between males and females. It is suggested that in ophidian cardiovascular studies the animals be grouped based upon their sex.
ABSTRACT
In this study we co-administered melittin along with HBsAg/alum vaccine to investigate if it helps elicitation of Th1/Th2 response. Hepatitis B virus [HBV] infection is a life-threatening liver infection, which can lead to chronic liver disease. Vigorous T cell responses are stimulated at acute, self-limiting HBV infection, while chronic HBV infection elicits very weak T cell responses. The prevalence of HBV infection has been decreased by the approved vaccination approach using recombinant HBs antigen [HBsAg] and alum i.e. HBV vaccine. Alum, a strong Th2 stimulator, is usually used as adjuvant to increase HBsAg immunogenicity. The present vaccine does not induce protective and/or prophylactic immune response in some groups. Melittin, major active component in the venom of honeybee, induces Th1 development. Experimental mice were immunized with melittin plus hepatitis B vaccine on day 0 following by two booster doses with the same injections. Lymphocyte proliferation, IFN-gamma, and IL-4 level, total antibody and isotyping of IgG1, IgG2a IgG2b, and IgM were measured using ELISA. Administration of melittin and HBV vaccine had no effect on lymphoproliferation and total antibody responses, but increased IFN-gamma response and induced Th1 response. The present study proposed that administration of melittin along with conventional vaccine shifts T cell responses towards Th1/Th2 dominated with Th1 response. The resultant immune response leads to activation of both cell-mediated and humoral immune responses, both of which required for clearance of HBV infection.
ABSTRACT
Objective: To evaluate a) heart place and tail length, b) their correlations with other biometrics, c) sexual differences in those features, in regard to cardiovascular system in a number of snakes from Iran, about which there is little information. Methods: We studied the fresh mortalities of snakes including 14 Naja oxiana (N. oxiana), 23 Macrovipera lebetina, and one male Montivipera latifii acquired from the Serpentatium of Pasteur Institute of Iran. In this respect, each specimen first was weighted, and then its ventral side of the integument was incised, and heart place was measured. Subsequently, other biometrical features such as total length, TAL, and snout-vent length were measured. Results: The results showed that heart place in N. oxiana, Macrovipera lebetina and Montivipera latifii was about 18%, 32% and 30%, and also TAL constituted about 16%, 11% and 7% of total body length, respectively. Moreover, females indicated anterior heart place and shorter tail than males. Furthermore, the measures and correlations indicated few differences between N. oxiana and typical terrestrial species. Conclusions: The results denoted that in order to overcome hemocirculatory perturbations in vertical orientation while hooding and head raising behavior, N. oxiana need to have more important features than short heart to head distance and long tail. In addition, it gave the sexual differences in heart place and tail length between males and females. It is suggested that in ophidian cardiovascular studies the animals be grouped based upon their sex.
ABSTRACT
Background: Gastric cancer [GC] is one of the most common cancers worldwide and in Iran. Conventional therapies are surgery and chemotherapy. Current studies are evaluating natural compounds in inhibiting growth of cancer cell. In this study isolated peptide melittin with 26 amino acids from bee venom and its impact on the viability and proliferation of gastric cancer cells was investigated
Methods: At first melittin was purified from honeybee venom using a reversed-phase high performance liquid chromatography [RP- HPLC] and C18 column. In order to investigate whether melittin, a 26 amino acids peptide which is the main components of honeybee venom, inhibits proliferation of human gastric adenocarcinoma cell line [AGS cells], MTT [[3-[4, 5-dimethylthiazol-2-yl]-2, 5- diphenyltetrazolium bromide] assay was performed. Hemolytic assay carried out in order to confirm the biologic activity of the isolated melittin. AGS cells were plated in a 96-well plate and treated with serially diluted concentrations of melittin for 6 and 12 hours. The mortality of the cells was measured via MTT assay at 540 nm
Results: The obtained chromatogram from RP-HPLC showed that melittin comprises 50% of the studied bee venom. SDS-PAGE analysis of melittin fraction confirmed purity of isolated melittin. Hemolytic activity assay indicates that isolated melittin shows a strong hemolytic activity [HD50=0.5]. MTT assay showed that melittin strongly inhibits proliferation of gastric cancer cells at concentrations more than 2microg/ml. This inhibitory effect is dependent to melittin concentration and incubation time
Conclusion: This study provides evidence that melittin inhibits proliferation of the gastric cancer cells. Results showed that isolated melittin from honey bee venom have cytotoxic effect on AGS cell line with a trend of increasing cytotoxicity with increasing concentration and incubation time
ABSTRACT
Ritalin has high tendency to be abused. It has been the main indication to control attention deficit hyperactivity disorder. The college students may seek for it to improve their memory, decrease the need for sleep [especially during exams], which at least partially, can be related to serotonergic system. Therefore, it seems worthy to evaluate the effect of Ritalin intake on mature brain. There are many studies on Ritalin effect on developing brain, but only few studies on adults are available. This study was undertaken to find Ritalin effect on serotonin transporter [SERT] density in medial frontal cortex [MFC] of mature rat. Thirty male Wistar rats were used in the study. Rats were assigned into five groups [n = 6 per group]: one control, two Ritalin and two vehicle groups. Twelve rats received Ritalin [20 mg/kg/twice a day] orally for eleven continuous days. After one week of withdrawal and another two weeks of rest, in order to evaluate short-term effects of Ritalin, six rats were sacrificed. Another six rats were studied to detect the long-term effects of Ritalin; therefore, they were sacrificed 12 weeks after the previous group. The immunohistochemistry was performed to evaluate the results. Immunohistochemistry studies showed a higher density of SERT in both 2 and 12 weeks after withdrawal from Ritalin intake in MFC of rat and there was no significant difference between these two groups. Our findings demonstrated both short- and long-term effects of Ritalin on frontal serotonergic system after withdrawal period
Subject(s)
Male , Animals, Laboratory , Serotonin , Rats, Wistar , Frontal Lobe , Cerebral CortexABSTRACT
Sequence analysis and phylogenetic study of hemagglutinin [HA] gene of H9N2 subtype of avian influenza virus isolates [outbreaks of 1998-2002] in Tehran province [Iran] were studied. Two sets of forward and reverse primers in highly conserved regions, based on sequences of HA gene in Genbank, were designed. PCR products of a 430-bp fragment of 16 isolates were sequenced and then were aligned with the reported sequences in Genbank. Nucleotide sequence comparisons of HA gene from Iranian isolates showed 97-99% identity within the group, and 98% homology with the two isolates [A/Parakeet/Narita/92A/98 [H9N2]] and [A/Parakeet/Chiba/1/97 [H9N2]] from Pakistani parakeets imported to Japan. On the basis of phylogenetic evidence, it is proposed that the emergence of H9N2 avian influenza infection in Iran originated in Pakistan, and it was due to low quarantine measures in the international boundaries. Due to the high percentage of H9N2 homology isolates of Iran with other isolates, namley A/quail/HongKong/G1, in Genbank and based on published reports for high similarity with infecting human H5N1 isolates, it seems that the potential of Iranian avian influenza isolates to infect human should be considered