NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2016 | Volume:6 | Issue:22|Papers Count:16

Aim and Background: Non-tuberculous mycobacteria have long been identified as capable of causing human disease. Several reports have suggested increasing trend to available treatment regimes. The aim of this study was to evaluate antibiotic susceptibility of clinically significant NTM using standard micro broth dilution test.Material and methods: The antimicrobial susceptibility testing was performed following National Committee for Clinical Laboratory Standards (NCCLS) method for 88 clinical isolates of slowly growing mycobacteria (SGM) and 154 clinical isolates of rapidly growing mycobacteria (RGM).Results: Among the 40 isolates of M. kansasii, all were susceptible to Ethambutol, Isoniazid, Clarithromycin, Moxifloxacin and Linezolid. The isolates also were resistant to Doxycycline and 50% isolates were resistant to Rifampicin and Ciprofloxacin.Conclusion: The presence of high variations in susceptibility among clinically important NTM to the currently available antimicrobial agents confirms the need for accurate identification as well as performing standard susceptibility testing of any clinically important isolate.

Aim and Background: The envelope glycoprotein E2 in Hepatitis C virus is required for host-cell entry. The nucleotide sequence diversity in carboxyl-terminus (C-terminal) of this gene results in induction of neutralizing antibody that makes this gene an important target for vaccine development studies. In this study we tried to make a recombinant pET28a (+) containing full length of the E2 gene from genotype 2a (JFH1) followed by being transformed into the E-coli (DH5a).Material and Methods: Full-length of HCV E2 region from genotype 2a (JFH1) was amplified by PCR reaction using the specific primers including the restriction sites for EcoR1 and Nde1. The confirmed PCR product was cloned into the pET28a (+) vector and transformed into the E-coli (DH5a) using the heat shock method.Results: The HCV E2 gene was successfully cloned into the pET28a (+) vector in full length size. Transformation of the E-coli strain DH5a by this recombinant vector was confirmed.Conclusion: Transformation of the E-coli (DH5a) using the recombinant vector pET28a- HCV 2a (JFH1) E2 gene provides useful tools for further expression of this gene in full length size in different expression system.

Aim and Background: Proteases is family of enzymes which has valuable commercial applications due to their very crucial physiological roles. Alkaline proteases is produced by Bacillus species which are of particular importance because of their thermal stability at different pHs. This study aimed to evaluate and compare two methods of package cultivation and semiconductor in immersed culture medium for the production of alkaline protease enzyme using 2, 10 and 20 percent of glucose substrate.Materials and methods: Bacteria of the genus Bacillus were isolated from alkaline soils of Guilan province and after screening for enzyme production being simultaneously evaluated in both. closed and semi-continuous culture methods.Results: Comparison of two methods showed that enzyme production in semi-continuous culture was 2.3 times more than package culture using 2% glucose (0.207U / ml vs. 0.088 U / ml) indicating that the former is more effective.Conclusion: isolated Bacillus strains could produce alkaline protease enzyme in both culture methods, but enzyme activity in semi-continuous culture was 3.2 times higher compared to packageculture.

Aim and Background: The recent discovery about the “miRNA expression is frequently altered in cancer,” and since it is now the second leading cause of cancer mortality in human society, it has opened a new direction of research. In the past several years, a very different approach to the development of various methods for the detection of miRNA biomarkers has been used. Over the past decade, an enormous progress has been made in the use of nanotechnology for molecular diagnostics.Materials and Methods: For the preparation of gold nanoparticles, a certain proportion of sodium citrate and HAuCl4 3 H2O were combined with each other. The mean diameter of the gold nanoparticles was calculated using the Zeta sizer. The ratios of specific oligonucleotide were added to the nanoparticles, nanoprobes made were exposed to target sequences, and the fluctuations of surface plasmons were investigated by UV-visible spectrometer.Results: The mean diameter of the gold nanoparticles synthesized was measured by Zeta sizer about 20 nm, respectively. The optimal concentration of oligonucleotide for binding to nanoparticles was about 5pM measured by UV-visible spectrometer. The detection limit of this method for the synthesis of a single strand of miRNA with the 23mer, is about 5pM.Conclusion: The study showed that gold nanoparticles prepared by citrate reduction method for the detection of miRNA biomarkers by using SPR properties of gold nanoparticles is possible.

Background: To study the effect of irrigation and zinc nutrition on yield and essential oil production of (Salvia officinalis L.), a field experiment was conducted at the College of Agriculture, Islamic Azad University of Khoy during spring 2013.Material and methods: The experiment was conducted as a factorial based on complete randomized block design with four replications. The effects of irrigation in four levels including 100, 80, 60, 40 % (FC) and zinc nutrition in four levels including 0 (control), 2.5, 5, 10 mg/kg (ZnSO4) were investigated.Results: The results showed that the effect of irrigation on plant height, number of leaves, stem diameter, fresh weight and dry weight were significant (p£1%). The highest oil yield (20.91 kg/ha) was obtained with 100% humidity. The results also showed that zinc spraying on the height, number of branches, leaves, internodes, fresh and dry weight, and percentage of essential oil was significant (p£0.01). Zinc Spraying and watering on only the main stem diameter and plant height was also significant (p£0.05). The highest essential oil yield (15.81) was obtained in 2.5 mg/kg (ZnSO4).Conclusions: According to our results it is recommended to increase the essential oil yield using 100% humidity and 2.5 mg/kg (ZnSO4) in (Salvia officinalis L.).

Aim and Background: Hypericin is a photosensitive Naphthodianthrones with photodynamic activities. In recent years, the pharmacology studies have shown that extracts and compounds obtained from Hypericum perforatum have some properties such as: antidepressants, anti-cancer, anti-antioxidant, anti-bacterial and anti-microbial and recently, there is great attention to this herb due to its effect for treatment of AIDS. In this work, the chemical stimuli effects of methyl-jasmonate and salicylic acid in medium stage (MS) on Hypericin production stimulation during a 42-day period was examined.Material and Methods: Stem and leaf explants were isolated from planted sterile seedlings in medium and were transferred to MS condition including methyl-jasmonate stimulus with concentration of (0, 50, 100, 150, 250 and 500 micro-Molar) and salicylic acid stimuli with concentration of (0, 50, 100, 150, 200 and 400 micro-Molar) and then were kept in temperature about 21±3oC in 16 hours lighting and 8 hours dark light condition. The methanol extracts of this plant were analyzed with HLPC apparatus for studying the changes in Hypericin amount.Conclusion: The results showed that the concentration of 100mM methyl- jasmonate produces more Hypericin (%34, 66) than other producers. Also these results showed that salicylic acid stimuli have no tangible effect on Hypericin production and using concentrations more than 100 mM of salicylic acid causes destruction of plant tissue cultures of Hypericum perforatum.Results: The chemical stimulus, methyl -jasmonate is more effective in Hypericum production of plant Hypericin tissue culture. The chemical irritant effect of salicylic acid did not significantly increase the amount Hypericin in the plant Hypericum.

Aims and Background: Different studies have shown that despite the expanding number of antifungal agents, death rate caused by Aspergillus species has been increased during the recent decades due to drug-resistance occurrence, increased minimum inhibitory concentration (MIC) and cross-resistance among the isolated species. Regarding the lack of effective response to conventional treatments and antifungal susceptibility patterns of the most common isolated Aspergillus species.Material and methods: In this study, during 9 months; 20 clinical isolates and 20 environmental strains were studied. In the next stage, drug sensitivity test was done according to the NCCLS-M38P method. A fungal suspension was made with 1-4*104 cfu/ml cellular range using 530nm spectrophotometer. Serial dilution of Itraconazol, Voriconazol (0.015-16mg/ml) and Caspofungin (0.007-8mg/ml) were performed and MIC was determined after 48Hr of incubation on 35oc.Results: In studied strains we did not find any drug resistance gene. MIC range for Aspergillus flavus in Itraconazol was determined as 0.125-2 mg/ml which MIC50= 0.5 and MIC90=1mg/ml. also for vericonazol range was 0.25-2 mg/ml and MIC 50=0.5 and MIC90=1 determined and caspofungin 0.0131-0.125 mg/ml and MIC50=0.063 and MIC50=0.063 and MIC90=0.125.Conclusion: According to MIC range for Itraconazol, voriconazol and caspofungin, the studied aspergillus flavus strains were drug sensitive; also, we did not find any drug resistance strain.

Aim and Background: Nowadays, using nanocarriers has increased in drug delivery industry. Niosome is one of the nanocarriers with non-ionic surfactant which increases the stability of drug in body. Since, using cisplatin in the treatment of cancers has some side effects, it was nanoniosomed.Materials and Methods: Nanoniosomes were synthesized by reverse phase evaporation and ether injection methods. The mean diameter of particles was determined using Zetasizer. The encapsulation and drug loading efficiency of two formulations which were prepared from both reverse phase evaporation and ether injection methods was measured by spectrophotometric method. Drug release pattern was examined by dialysis in a period of 48 hours. Cytotoxicity effect on nanoniosomal cisplatin was determined on C6 cell line by MTT assay.Results: The mean diameter of particles was synthesized by reverse phase evaporation and ether injection methods was determined 242.1±5.10 nm and 218.6±3.42 nm, respectively. The encapsulation and drug loading efficiency of two formulations were measured 43.6±4.59%, 52.8±3.22%; 4.36±0.08% and 5.28±0.17%, respectively. The results of drug release showed that released drug from prepared formulation using reverse phase evaporation was 97.53±0.55% and using ether injection was 81.23±1.24%. In addition, the results of cytotoxicity showed that the cytotoxicity of prepared formulation using ether injection more than prepared formulation applying reverse phase evaporation.Conclusion: This study showed that using reverse phase evaporation to synthesize nanoniosomal cisplatin is more appropriate method than ether that of injection method.

Aims and Background: Since the birth of the first baby using In vitro fertilization (IVF) in 1978, huge changes have been established in the field of reproductive biology and infertility treatment. Although IVF is considered as a useful technique, but studies have shown that during IVF, some stresses induced by osmosis, oxygen, temperature and pH changes in ectopic conditions can cause morphological, genetic and epigenetic changes in embryos derived from IVF compared to the In vivo. The purpose of this study was to assess the effect of IVF in embryo quality regarding the survival and hatching rates and the expression level of Oct4 and Nanog genes. These genes are effective in the pluripotency of blastocyst inner cell mass.Materials and Methods: In this experiment 3.5 to 4.5 days old mouse blastocysts were studied in two groups. In treated group, In vitro blastocysts were obtained by passing 100 hours after In vitro fertilization of sperms and oocytes. Fresh blastocysts were also considered as control group. Survival and hatching rates and also Oct4 and Nanog genes expression level were evaluated using Real Time PCR technique in both groups.Results: Although the hatching rate of blastocysts decreased significantly in treated group (67.73±5.80 vs. 86.88±10.40%), the survival rate was remained 100% as well. In addition Oct4 and Nanog genes expression level was significantly increased in, In vitro blastocyst compared to the control group.Conclusion: According to the results, in vitro fertilization can change gene expression and quality in the embryo.

Aim and Background: Formation of the blood clot inside a blood vessel is known as thrombose or thrombosis. This complication usually appears through the blood platelets. The tendency to clot originates from external and genetic factors which in turn, is a result of changing in the clotting mechanism.With the aim of applying a method known as ‘’Taq man Allele- Specific Real-Time’’ for diagnosis and screening of V617F mutation in JAK2 gene, this study was carried out in those patients being susceptible to thrombosis.Materials and Methods: In this study, through referring to the Pathobiology Laboratory of RASHT city situated in Guilan province, 110 perfect bloods from the afflicted individuals and 105 perfect bloods from the ordinary individuals (lacking past record of the disease) were provided out of the referrers in order to study the thrombosis factors and, after DNA extraction from the whole samples, required measures to study presence of V617F mutation in JAK2 gene were taken by a method known as Taq man Allele-Specific Real-Time PCR, then, using statistical methods, the results were analyzed.Results: After confirming the accuracy of the desired technique applied on the control samples, the results of screening this method in the population of individual referring to study the factors of thrombosis, including factors of 2 and 5, presence of this change was suggested. Also, none of the V617F mutation in JAK2 was identified by screening on the samples of the control population.Conclusion: Our findings showed that the applied technique is sufficiently capable of diagnosing the V617F mutation in JAK2 gene. Also, lack of identification of V617F mutation in JAK2 gene in the population of those referring to the Pathobiology Laboratory for thrombosis in order to investigate factors of 2 and 5 showed that V617F mutation in JAK2 gene in the studied population is not propounded as one of the thrombosis factors.

Aims and Background: Diabetes mellitus is a chronic disease caused by lack of insulin secretion or inability of cells to use insulin. Various methods are used to treat the disease; however they are associated with many problems. Therefore, access to novel methods of treatment and prevention of the disease is of great importance. This study investigated the impacts of a native probiotic of Iran and cumin plant extract on reducing blood sugar levels in diabetic rats.Materials and Methods: Thirty-five male Wistar rats aged 4 weeks, with same weighs of 110-130 g were selected. After 2 weeks of adaptation, they were made diabetic by streptozotocin and divided randomly into normal control, diabetic control, probiotic receiving diabetic, cumin receiving diabetic, and probiotic plus cumin receiving diabetic groups. The probiotic used was Bacillus subtilis JQ61819, purchased as lyophilized powder (1×109 cfu/g) from Takgene Co., and cumin extracts were purchased from Barij Essence pharmaceutical Co. The rats were daily fed with the probiotic and cumin through gavage using a needle. After the treatment period, the rats were anesthetized and blood samples were taken from their hearts, and sugar and lipids were measured in serum using appropriate kits.Results: Bacillus subtilis and cumin extract lowered blood sugar by 61% and cholesterol by 44% and increased triglycerides by 14% in diabetic Wistar rat models.Conclusion: The use of indigenous probiotic Bacillus subtilis alone or in combination with cumin extract may be useful in controlling blood sugar in patients with diabetes.

Aim and background: One of the world’s important pharmaceutical resources is Greater Celandine. Since the genetic diversity of this plant is not assessed in the global studies, this study investigates the genetic diversity in Greater Celandine via molecular markers ISSR in 14 ecotypes.Materials and methods: DNA was extracted from young leaves of all the genotypes using the CTAB method. The ISSR products were scored for the presence (1) and absence (0) of each primer. Clustering result of molecular data was divided distinctly into 7 quite genetic groups.Results: primers (CA) 8G provided the most polymorphism (92.8) among the primers. The lowest genetic similarity was 0.16 between two samples Shirgah (C16) and Siahkolah (C30).Conclusion: The results of classified cluster clearly indicated that there is no relation between molecular diversity and geographical variety. It seems that it might be possible that one sample of a geographical region is migrated to another region being distanced from its primary source.

Aim and Background: Chromosomal information of different species, in addition to improving classification and biological recognitions; it can be effective in breeding and genetic studies.Material and Methods: Bred stocks were preyed from the east and west of Jask Harbour. Different growth stages were stained by Cholchicine and hypotonic KCL in different concentration and duration. Metaphase chromosome spreads were studied based on Campos-Ramos and Crash tissue methods and stained by Giemsa and distilled water.Results: The testis tissue showed the best obtained spread. The number of diploid chromosomes (2n=86) was confirmed by the modal haploid chromosome number (n=43) in which 27 and 16 chromosome pairs were counted in meta/submetacentric and acro/telocentric, respectively.Discussion: despite observed chromosome variations in karyotyping further Penaeidae family, the achieved diploid chromosome number of this study (2n=86) exits within the range of diploid chromosome numbers previously reported for other species of shrimp (86 to 92).Conclusion: treatments of testis tissue, staining by Giemsa 10% and crashing tissue method recognized as the best methods for karyotyping and chromosome spread analysis of Penaeidae family.

Aim and Background: Silver nanoparticles have many applications in different industries. In original methods nanoparticles are synthesized by chemical methods. While, in this research we synthesized Silver nanoparticles (AgNPs) within Poly (N-vinyl-2-pyrrolidone) (PVP) hydrogel matrix by Electron beam irradiation.Materials and Methods: For hydrogel preparation, PVP (7% wt) was dissolved in distilled water and heated at 80oC under stirring. Then 70,100 and 200 ppm amount of AgNO3 solution was added into solution. The samples were then irradiated with 33 kGy. Ag/PVP hydrogel was characterized using UV-Vis spectroscopy and transformed infrared spectroscopy (FTIR), Atomic force microscopy, swelling ratio, gel content and antibacterial activity test.Results: The Plasmon Absorption spectra of Ag/PVP nanocomposite confirmed the formation of AgNPs. The Atomic Force microscopy (AFM) analysis of Ag/PVP nanocomposite showed AgNPs in the range of 5-10 nm. FTIR spectrum indicated that, the coordination between N in PVP and silver was the main reaction. Ag/PVP nanocomposite and pure PVP exhibited the same values of swelling ratio and Gel fraction. Antimicrobial test indicated Ag/pvp hydrogel has antimicrobial properties.Conclusion: In this research, it was shown the electron irradiation is a suitable technique for synthesis of AgNPs into hydrogel matrix and insitu synthesis of AgNPs simultaneously with sterilization, making this method very attractive for biomedical applications.