Original Article

Cloning and Expression of Recombinant Nucleoprotein of Influenza H1N1

Somaie Tavakoli, Mojgan Bandehpour, Zeinab Soleimanifar, Mehdi Goudarzi, Bahram Kazemi

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 53-56

Background: Influenza virus is the major cause of lower respiratory tract illnesses on the worldwide. Vaccination can be an effective tool to prevent its outbreak. Highly conserved viral nucleoprotein is an effective vaccine candidate to provide heterosubtypic immunity, offering resistance against various influenza virus strains.

Materials and Methods: In present research NP gene was inserted in pET-22b expression vector. New construct (pET-22b/NP) was transformed into E. coli BL21 (DE3) strain and the expression of nucleoprotein was induced by IPTG. It was analyzed by SDS-PAGE and confirmed by Western blotting.

Results: Western blotting confirmed the expression and production of recombinant Influenza nucleoprotein.

Conclusion: These results suggest that the codon-optimized influenza A virus NP gene can be efficiently expressed in E. coli.

Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin

Mehdi Azad, Mehdi Goudarzi, Ali Dehghanifard, Mousa Vatanmakanian, Mehdi Sahmani

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 57-62

Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin.

Materials and Methods: After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively.

Results: Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique.

Conclusion: It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation.

Background: The rodent somatosensory barrel cortex is an ideal model to examine the effect of experience-dependent plasticity on developing brain circuitry. Sensory deprivation such as whisker deprivation may affect neuroanatomical aspects of the brain during developmental processes. The present study designed to investigate the possible effects of whisker deprivation on the morphometric characteristics of NADPH-d positive neurons in the barrel field cortex of adolescent rats.

Materials and Methods: Pups were divided into the intact (n=4) and whisker-deprived groups (n=4). In whisker-deprived group, the total whiskers of subjects were trimmed every other day from postnatal day (PND) 0 to PND 60. NADPH-d histochemistry reaction was processed to quantitatively analyze the feature of NADPH-d containing neurons of barrel cortex.

Results: Our results showed that the number of NADPH-d positive neurons remained unchanged in whisker-deprived group compared to controls. The mean soma diameter, dendritic length and the number of 3rd order processes were significantly decreased in the whisker-deprived rats (p<0.05).

Conclusion: Our results indicate that postnatal whisker deprivation possibly alter NADPH-d/NOS neuronal features in the barrel cortex. The functional implications of these data may relate the plasticity of synaptic receptive field and developmental brain circuits.

Background: Acanthamoeba-related disease have a poor prognosis according to many previous studies. Thus researches regarding biochemical and molecular aspects of this organism are a high priority. To this end achieving high amount of amoebae in culture is the first step for such studies. The main aim of the present research was to address the usage of TYI-S-33 (Tripticase, Yeast extract, iron-serum) medium as an enrichment component for achieving high and fast growth of trophozoites in agar culture medium within 24 hours.

Materials and Methods: Overall, 10 Acanthamoeba strains were cultured, cloned and genotyped and the cultures were then enriched with addition of TYI-S-33 medium. Amoebae growth was then monitored daily. Ten plates also were used without addition of TYI-S-33 medium.

Results: The result of the present research revealed that addition of TYI-S-33 medium is a promising approach for obtaining 100% trophozoites within 24 hours of culture.

Conclusion: To the best of our knowledge this is the first report of successful achieving high amount of trophozoites within short time that able researchers to arrange molecular and biochemical assays

The Prevalence of Ureaplasma Urealyticum and Mycoplasma Genitalium in Patients with Prostate Cancer in Shohada Hospital in Tehran, Iran

Gita Eslami, Hossein Goudarzi, Neda Baseri, Zohreh Ghalavand, Arezou Taherpour, Haniye Zhaam, Roghayeh Samadi, Eznolah Azargashb

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 73-78

Background: Prostate cancer (PCa) is an important health problem in the aging male population in the world. It is the third most common cancer in the world. Despite of its importance, relatively little is known about its etiology. Sexually transmitted infections (STI) and urogenital pathogens such as Mycoplasma and Ureaplasma, have been proposed as a risk factor for prostate cancer development. This study aimed at detecting the prevalence of Ureaplasma urealyticum (U. urealyticum) and Mycoplasma genitalium (M. genitalium) in PCa and the controls group with benign prostate hyperplasia (BPH) in Shohada hospital.

Materials and Methods: A total of 124 paraffin-embedded prostate tissues (62 PCa patients and 62 controls with BPH) were included in this study. The subjects'specimens were investigated by the polymerase chain reaction method for the presence of U. urealyticum and M. genitalium DNA.

Results: U. urealyticum was detected by standard PCR in 1.61% of the 62 PCa patients and there was no DNA U. urealyticum in the 62 controls with BPH. No M. genitalium was detected by standard PCR in the prostates of 124 paraffin-embedded prostate tissues.

Conclusion: According to our results, there is no association between M. genitalium and U.urealyticum with PCa. We recommend further studies using a large sample to determine role of Ureaplasma and Mycoplasma in PCa because understanding the role of infectious agents on PCa might be useful for developing new therapeutic approaches and prevention of PCa.

Background: Streptococcus group B (GBS) or Streptococcus agalactiae is typically associated with neonatal disease and infection in pregnant women. Mortality of GBS sepsis in neonates is over 50% and is particularly high in preterm infants. GBS also causes invasive infection in pregnant and non-pregnant women including urinary tract infection (UTI). Penicillin-derived antibiotics remained as choice drugs for treatment of GBS infection; however, Erythromycin and Clindamycin are useful in cases of allergic to Penicillin. The aim of this study was to investigate the resistance to Erythromycin and Clindamycin, especially inducible Clindamycin resistance, in GBS isolated from urinary samples of women who attended medical offices in Tehran, Iran.

Materials and Methods: This study was conducted on 5000 urine samples from Jan. 2011 to Oct. 2012 that 104 GBS were isolated. The isolates were identified as GBS using laboratory criteria. Antimicrobial susceptibility test was done by Erythromycin disk 15µg and Clindamycin disk 2µg for observation inducible resistant D-zone test by double-disk diffusion method with Erythromycin and adjacent Clindamycin.

Results: Among the 5000 urine samples 104 (2.08%) were Beta hemolytic GBS. Of the 104 isolated GBS, 22 (21.2%) were resistance, 24 (23%) were intermediate, and 58 (55.8%) were susceptible to Erythromycin; however, 24 (23%) were resistance, 5 (4.8%) were intermediate, and 75 (72.2%) were susceptible to Clindamycin. Of the 22 Erythromycin-resistant isolates, 10 (9.5% in total GBS isolated) displayed the D zone; it means they have inducible Erythromycin resistant to Clindamycin.

Conclusion: Various studies in other countries report lower rates of inducible Clindamycin resistance; it indicates the use of more macrolides in the treatment of UTI.

Bacterial Infection of Pacemaker in Patients with Endocarditis

Fariba Fayaz, Sara Rahmati roodsari, Latif Gachkar, Bita Pourkaveh

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 84-88

Background: The advancement of technology in recent decades has been lead to use the electrophysiology cardiac devices. Although these devices are used increasingly, but the frequency of subclinical infection is unknown. We investigate bacterial infections due to implantable cardioverter defibrillator (ICDs) in patients with endocarditis.

Materials and Methods: Population of the study was considered among all adult patients in whom the cardiac electrophysiology device was removed. Associated infection endocarditis defined by the Duke criteria. 35 pacemakers (PM) were aseptically removed from these patients during January 2012 to November 2014. Intraoperative swabs from the different part of devices were collected, cultured in BHI (Brain Heart Infusion Broth) and then bacterial classical cultures were done under aerobic and anaerobic conditions. Biochemical and differential media were used to detect the bacteria species. Data analysis was performed by using SPSS version 16 software.

Results: 13 cases of 35 patients with endocarditis diagnosed by modified Duke Criteria and removed pacemaker had positive culture. Of the 13 cases with infection 43% were identified as gram positive and 57% had gram negative bacteria.

Conclusion: Based on our study and similar studies, bacteria can colonize in electrophysiology devices which can lead to bacterial infections.


Fibrinolytic Activity of Recombinant Mutant Streptokinase

Mahboobeh Mobarrez, Mojgan Bandehpour, Massoud Salehmoghadam, Bahram Kazemi

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 89-93

Background: Streptokinase is a bacterial protein produced by different beta hemolytic streptococci and widely used in thrombolytic treatment. The main disadvantage of using streptokinase is antibody formation which causes allergic reaction to neutralize effects of streptokinase therapy. Aim of this study was investigate of recombinant mutant streptokinase fibrinolytic activity.

Materials and Methods: In this study recombinant mutant streptokinase without 42 amino acids from the C terminal region was purified by affinity S-Tag column chromatography and its fibrinolytic activity was studied.

Results: The concentration of expressed and purified protein was 10 mg/ml. Its enzyme activity was assayed using zymography, radial caseinolytic activity and fibrin plate test methods and estimated quantitatively by casein digestion method compared to a commercial form.

Conclusion: It was found that this product had the more volume and more enzymatic activity.

Case Report

Familial Dysautonomia, Report of 3 Cases from Iran and a Discussion about Their General and Anaesthesia Care

Badiozaman Radpay, Mahtab Poor Zamany Nejat Kermany, Ali Asghar Halimi-Asl

Novelty in Biomedicine, Vol. 3 No. 2 (2015), , Page 94-98

Background: Familial Dysautonomia (FD) is a rare hereditary syndrome which is an autosomal recessive trait that typically affects Jewish children. Important signs and symptoms of the disorder include; diminished pain perception, absence of overflow tears, hypotonia, fainting cardiac arrhythmias and autonomic crisis.

Cases Report: In this article we reported 3 cases of FD syndrome which had presented for surgical operation followed by a discussion about general care of these patients as well as Anesthesia considerations.