Biotechnology and Health Sciences Biotechnology and Health Sciences Biotech Health Sci http://www.Biotech-health.com 2383-0271 2383-028X 10.5812/bhs en jalali 2017 5 27 gregorian 2017 5 27 1 3
en 10.17795/bhs-25362 Epidemiological Study of Non-Melanoma Skin Cancers Qazvin Province, Iran Epidemiological Study of Non-Melanoma Skin Cancers Qazvin Province, Iran research-article research-article Background

Non-melanoma skin cancers (NMSC) are the most common cancers worldwide. These cancers are not accompanied by high mortality however they lead to various complications. Evaluation of NMSC predisposing factors could result in preventive measures, improvement of quality of life and reduction of medical costs.

Objectives

This study aimed to determine predisposing factors of non-melanoma skin cancers in the province of Qazvin (Iran).

Materials and Methods

A total of 484 proven cases of NMSC were evaluated for demographic characteristics, clinical findings, pathological type, location of lesion, and the existence of known or possible predisposing factors such as skin type, exposure to sunlight, and family and drug history within a ten-year period from 2001 to 2011. Data were recorded in a questionnaire through interview and clinical examination of patients by a physician and a dermatologist, respectively. Data were analyzed using the SPSS version 16 software and statistical tests including t-test and chi-square tests.

Results

Of the 484 patients, 294 (60.7%) were male and 190 (39.3%) were female. Most cases of carcinoma were found in basal cells (77.7%) and the rest of squamous cells. The most common sites of involvement were the head and scalp. The most frequent predisposing factors were working in the open air (70.7%), history of previous radiation for treatment of tenia capitis (26.1%), and chronic skin diseases such as burn eschar, chronic lesion, fistula and actinic keratosis (35.7%). The most prevalent Fitzpatrick skin phenotypes were II or III (75.4%). There were significant correlations between the incidence of NMSC and hookah smoking and oral contraceptive pill (OCP) consumption.

Conclusions

Skin phenotypes II and III were the most common types found in the present study and this could be due to the higher frequency of these phenotypes among the study population. The lower incidence of NMSC in areas other than the neck and scalp could be associated with the importance of covering style used by the population under study. Known risk factors for the incidence of NMSC were also observed in the present study. A history of radiotherapy for treatment of tenia capitis was observed in a considerable percentage of patients, which could lead to the incidence of cancer several decades later. Further studies are needed to determine the role of hookah smoking and the use of OCP in the occurrence of NMSC.

Background

Non-melanoma skin cancers (NMSC) are the most common cancers worldwide. These cancers are not accompanied by high mortality however they lead to various complications. Evaluation of NMSC predisposing factors could result in preventive measures, improvement of quality of life and reduction of medical costs.

Objectives

This study aimed to determine predisposing factors of non-melanoma skin cancers in the province of Qazvin (Iran).

Materials and Methods

A total of 484 proven cases of NMSC were evaluated for demographic characteristics, clinical findings, pathological type, location of lesion, and the existence of known or possible predisposing factors such as skin type, exposure to sunlight, and family and drug history within a ten-year period from 2001 to 2011. Data were recorded in a questionnaire through interview and clinical examination of patients by a physician and a dermatologist, respectively. Data were analyzed using the SPSS version 16 software and statistical tests including t-test and chi-square tests.

Results

Of the 484 patients, 294 (60.7%) were male and 190 (39.3%) were female. Most cases of carcinoma were found in basal cells (77.7%) and the rest of squamous cells. The most common sites of involvement were the head and scalp. The most frequent predisposing factors were working in the open air (70.7%), history of previous radiation for treatment of tenia capitis (26.1%), and chronic skin diseases such as burn eschar, chronic lesion, fistula and actinic keratosis (35.7%). The most prevalent Fitzpatrick skin phenotypes were II or III (75.4%). There were significant correlations between the incidence of NMSC and hookah smoking and oral contraceptive pill (OCP) consumption.

Conclusions

Skin phenotypes II and III were the most common types found in the present study and this could be due to the higher frequency of these phenotypes among the study population. The lower incidence of NMSC in areas other than the neck and scalp could be associated with the importance of covering style used by the population under study. Known risk factors for the incidence of NMSC were also observed in the present study. A history of radiotherapy for treatment of tenia capitis was observed in a considerable percentage of patients, which could lead to the incidence of cancer several decades later. Further studies are needed to determine the role of hookah smoking and the use of OCP in the occurrence of NMSC.

Skin Cancers;Predisposing Factors;Radiation Skin Cancers;Predisposing Factors;Radiation http://www.Biotech-health.com/index.php?page=article&article_id=25362 Akram Beheshtiroy Akram Beheshtiroy Department of Dermatology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Dermatology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran Fatemeh Hajmanoochehri Fatemeh Hajmanoochehri Department of Pathology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Pathology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel/Fax: +98-2833348217 Department of Pathology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Pathology, Bouali Hospital, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel/Fax: +98-2833348217
en 10.17795/bhs-24733 Evaluation of Antioxidant, Antibacterial, and Antifungal Properties of Satureja hortensis Essential Oil Evaluation of Antioxidant, Antibacterial, and Antifungal Properties of <italic>Satureja hortensis</italic> Essential Oil research-article research-article Conclusions

The data of the study clearly indicated that the EO of S. hortensis has a strong antioxidant, antibacterial, and antifungal activity.

Results

Total phenolic content was determined by using Folin-Ciocalteu reagent (32.65 mg/g), which was presented as gallic acid equivalent in 1 g of sample. IC50 of EO and ascorbic acid in DPPH method were respectively 277.9 and 19.34 µg. Minimum inhibitory concentration of the EO of S. hortensis against Salmonella typhimurium, Listeria monocytogenes, Escherichia coli, and Bacillus cereus was respectively 2.5%, 2.5%, 5%, and 2.5%. The inhibition zone of EO in disk and agar well diffusion method showed that inhibitory zone on B. cereus was higher than that on S. typhimurium in both methods and B. cereus was more sensitive to EO. MIC to minimal fungicidal concentration (MFC) ratio of S. hortensis EO against Candida albicans, Candida parapsilosis, and Candida krusei in broth microdilution method were respectively 0.048%:0.048%, 0.024%:0.024%, and 0.012%:0.012%.

Objectives

The aim of this study was to investigate antimicrobial and antioxidant activity of essential oil (EO) of Satureja hortensis (Lamiaceae) that grows in Sabalan Mountain (Ardebil province, Iran).

Materials and Methods

This EO was tested in vitro against two bacterial species by disk and agar well diffusion methods and against four bacterial species and three Candida strains by broth microdilution method. Total phenol, flavonoid, and free radical scavenging activity of EO were evaluated.

Background

Satureja is a genus belonging to the aromatic plants of Lamiaceae family. The genus Satureja L. (Lamiaceae) comprises more than 30 species of aromatic herbs and shrubs, widely distributed over the Mediterranean region. The genus is represented by 14 species in Iran of which, eight are endemic. Many species of the genus Satureja are reported to have aromatic and medicinal properties. The leaves, flowers, and stems of this plant are used as herbal tea and in treatment of various ailments in traditional medicine.

Conclusions

The data of the study clearly indicated that the EO of S. hortensis has a strong antioxidant, antibacterial, and antifungal activity.

Results

Total phenolic content was determined by using Folin-Ciocalteu reagent (32.65 mg/g), which was presented as gallic acid equivalent in 1 g of sample. IC50 of EO and ascorbic acid in DPPH method were respectively 277.9 and 19.34 µg. Minimum inhibitory concentration of the EO of S. hortensis against Salmonella typhimurium, Listeria monocytogenes, Escherichia coli, and Bacillus cereus was respectively 2.5%, 2.5%, 5%, and 2.5%. The inhibition zone of EO in disk and agar well diffusion method showed that inhibitory zone on B. cereus was higher than that on S. typhimurium in both methods and B. cereus was more sensitive to EO. MIC to minimal fungicidal concentration (MFC) ratio of S. hortensis EO against Candida albicans, Candida parapsilosis, and Candida krusei in broth microdilution method were respectively 0.048%:0.048%, 0.024%:0.024%, and 0.012%:0.012%.

Objectives

The aim of this study was to investigate antimicrobial and antioxidant activity of essential oil (EO) of Satureja hortensis (Lamiaceae) that grows in Sabalan Mountain (Ardebil province, Iran).

Materials and Methods

This EO was tested in vitro against two bacterial species by disk and agar well diffusion methods and against four bacterial species and three Candida strains by broth microdilution method. Total phenol, flavonoid, and free radical scavenging activity of EO were evaluated.

Background

Satureja is a genus belonging to the aromatic plants of Lamiaceae family. The genus Satureja L. (Lamiaceae) comprises more than 30 species of aromatic herbs and shrubs, widely distributed over the Mediterranean region. The genus is represented by 14 species in Iran of which, eight are endemic. Many species of the genus Satureja are reported to have aromatic and medicinal properties. The leaves, flowers, and stems of this plant are used as herbal tea and in treatment of various ailments in traditional medicine.

Essential Oils;Antioxidants;Traditional Medicine Essential Oils;Antioxidants;Traditional Medicine http://www.Biotech-health.com/index.php?page=article&article_id=24733 Soghra Valizadeh Soghra Valizadeh Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Urmia University, Urmia, IR Iran Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Urmia University, Urmia, IR Iran Tayebeh Fakheri Tayebeh Fakheri Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran Razzagh Mahmoudi Razzagh Mahmoudi Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran; Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran. Tel: +98-4136378743, Fax: +98-4136378743 Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran; Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran. Tel: +98-4136378743, Fax: +98-4136378743 Farzad Katiraee Farzad Katiraee Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran Payman Gajarbeygi Payman Gajarbeygi Department of Public Health, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Public Health, Qazvin University of Medical Sciences, Qazvin, IR Iran
en 10.17795/bhs-24729 Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles research-article research-article Background

Cucumber mosaic virus (CMV) has isometric particles with a diameter of about 28–29 nm. A useful method to recognize regions from mutagenesis in the coat protein (CP) gene are important for particle assembly and the expression of viral in CP expression systems like bacteria such as E.coli or yeast as well as their assembly into virus-like particles (VLP).

Objectives

In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene.

Materials and Methods

In this study, the CMV-CP gene was released from a previously prepared cloning vector. Then, the CMV-CP was ligated into the expression vector. Sequencing was done by Marcrogen, Inc. (South Korea). A recombinant plasmid was transferred to E.coli isolate Rosetta. After inducing by isopropyl thiogalactosides, the molecular weight of the expressed protein was determined by SDS-PAGE. The extraction of proteins was done by NATURE method to see the possible presence of CMV-like particles.

Results

CMV-CP was detected by Western blotting by a CMV specific polyclonal antibody and conjugate. The protein extracted from the CP producing clone was studied under a JEOL 100-CXII transmission electron microscope with 100000× magnification at an acceleration voltage of 100 kV.

Conclusions

The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMV-like particle.

Background

Cucumber mosaic virus (CMV) has isometric particles with a diameter of about 28–29 nm. A useful method to recognize regions from mutagenesis in the coat protein (CP) gene are important for particle assembly and the expression of viral in CP expression systems like bacteria such as E.coli or yeast as well as their assembly into virus-like particles (VLP).

Objectives

In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene.

Materials and Methods

In this study, the CMV-CP gene was released from a previously prepared cloning vector. Then, the CMV-CP was ligated into the expression vector. Sequencing was done by Marcrogen, Inc. (South Korea). A recombinant plasmid was transferred to E.coli isolate Rosetta. After inducing by isopropyl thiogalactosides, the molecular weight of the expressed protein was determined by SDS-PAGE. The extraction of proteins was done by NATURE method to see the possible presence of CMV-like particles.

Results

CMV-CP was detected by Western blotting by a CMV specific polyclonal antibody and conjugate. The protein extracted from the CP producing clone was studied under a JEOL 100-CXII transmission electron microscope with 100000× magnification at an acceleration voltage of 100 kV.

Conclusions

The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMV-like particle.

Cucumovirus;Gene Expression;Recombinant Proteins;Virus Assembly;Electron Microscopy Cucumovirus;Gene Expression;Recombinant Proteins;Virus Assembly;Electron Microscopy http://www.Biotech-health.com/index.php?page=article&article_id=24729 Afshin Rostami Afshin Rostami Department of Plant Protection, University of Zanjan, Zanjan, IR Iran; Department of Plant Protection, University of Zanjan, Zanjan, IR Iran. Tel/Fax: +98-2632238529 Department of Plant Protection, University of Zanjan, Zanjan, IR Iran; Department of Plant Protection, University of Zanjan, Zanjan, IR Iran. Tel/Fax: +98-2632238529 Nemat Sokhandan Bashir Nemat Sokhandan Bashir Department of Plant Protection, University of Tabriz, Tabriz, IR Iran Department of Plant Protection, University of Tabriz, Tabriz, IR Iran Parviz Pirniakan Parviz Pirniakan Department of Agriculture, Payame Noor University, IR Iran Department of Agriculture, Payame Noor University, IR Iran Nahid Masoudi Nahid Masoudi Department of Plant Protection, University of Guilan, Rasht, IR Iran Department of Plant Protection, University of Guilan, Rasht, IR Iran
en 10.17795/bhs-24879 Evaluation of AmpC Gene Expression in Carbapenem Resistant Pseudomonas aeruginosa Strains Isolated from Severely Burned Patients With Secondary Infection in Hospitals Evaluation of AmpC Gene Expression in Carbapenem Resistant <italic>Pseudomonas aeruginosa</italic> Strains Isolated from Severely Burned Patients With Secondary Infection in Hospitals research-article research-article Background

Pseudomonas aeruginosa is an opportunistic pathogen mostly affecting hospitalized and immunodeficient patients. Considering the extent of attention paid to the issue of antibiotic resistance and the origin of such resistance and the growing number of treatment failures in patients with burn injury, it seems that investigating the degree of ampC gene expression could be beneficial for the treatment of patients with severe burn injury.

Objectives

The present study focused on identifying different antibiotic patterns, detecting carbapenem resistant Pseudomonas aeruginosa strains isolated from severely burned patients and investigating AmpC gene expression as one of the important mechanisms associated with drug resistance.

Materials and Methods

A total of 189 clinical isolates of carbapenem resistant Pseudomonas aeruginosa, isolated from patients with severe burn injuries, were identified by bacteriological methods followed by determination of their antibiotic resistance patterns by the standard protocol of Kirby–Bauer. The expression of ampC gene was determined by quantitative real-time PCR.

Results

Based on our findings, 94.2% of the isolates were resistant to imipenem, 99.5% were resistant to meropenem, and all were resistant to ertapenem. The level of ampC gene expression in carbapenem resistant Pseudomonas aeruginosa isolates, compared to standard carbapenem sensitive strains, was significantly increased.

Conclusions

Our data showed that the over-expression of ampC gene is an important mechanism of resistance in carbapenem resistant Pseudomonas aeruginosa strains isolated from severely burned patients with secondary infections.

Background

Pseudomonas aeruginosa is an opportunistic pathogen mostly affecting hospitalized and immunodeficient patients. Considering the extent of attention paid to the issue of antibiotic resistance and the origin of such resistance and the growing number of treatment failures in patients with burn injury, it seems that investigating the degree of ampC gene expression could be beneficial for the treatment of patients with severe burn injury.

Objectives

The present study focused on identifying different antibiotic patterns, detecting carbapenem resistant Pseudomonas aeruginosa strains isolated from severely burned patients and investigating AmpC gene expression as one of the important mechanisms associated with drug resistance.

Materials and Methods

A total of 189 clinical isolates of carbapenem resistant Pseudomonas aeruginosa, isolated from patients with severe burn injuries, were identified by bacteriological methods followed by determination of their antibiotic resistance patterns by the standard protocol of Kirby–Bauer. The expression of ampC gene was determined by quantitative real-time PCR.

Results

Based on our findings, 94.2% of the isolates were resistant to imipenem, 99.5% were resistant to meropenem, and all were resistant to ertapenem. The level of ampC gene expression in carbapenem resistant Pseudomonas aeruginosa isolates, compared to standard carbapenem sensitive strains, was significantly increased.

Conclusions

Our data showed that the over-expression of ampC gene is an important mechanism of resistance in carbapenem resistant Pseudomonas aeruginosa strains isolated from severely burned patients with secondary infections.

Pseudomonas aeruginosa;AmpC beta-lactamases;Drug Resistance Pseudomonas aeruginosa;AmpC beta-lactamases;Drug Resistance http://www.Biotech-health.com/index.php?page=article&article_id=24879 Fatemeh Attaran Rezaei Fatemeh Attaran Rezaei Department of Microbiology, Zanjan Branch Islamic Azad University, Zanjan, IR Iran Department of Microbiology, Zanjan Branch Islamic Azad University, Zanjan, IR Iran Taghi Naserpour Farivar Taghi Naserpour Farivar Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-9128801401, Fax: +98-2813324971 Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-9128801401, Fax: +98-2813324971 Masoumeh Aslanimehr Masoumeh Aslanimehr Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Reza Shapouri Reza Shapouri Department of Microbiology, Zanjan Branch Islamic Azad University, Zanjan, IR Iran Department of Microbiology, Zanjan Branch Islamic Azad University, Zanjan, IR Iran Akram Azimi Akram Azimi Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Pooran Johari Pooran Johari Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Cellular and Molecular Research Center, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran
en 10.17795/bhs-24683 Producing Probiotic Peach Juice Producing Probiotic Peach Juice research-article research-article Background

Probiotics have been used for dairy products such as yogurt and yogurt drinks, however cholesterol content and lactose intolerance are important drawbacks. Recently, consumption of non-dairy probiotic food especially for probiotic drink products has been intensified.

Objectives

This research was conducted to determine the suitability of peach as a raw material for producing probiotic peach juice by lactic acid bacteria.

Materials and Methods

Peach juice was inoculated with a 24-hour-old lactic acid bacteria culture and incubated at 30°C. Changes in the pH, titratable acidity, sugar content, and viable cell counts during fermentation under controlled conditions were monitored. Viability of lactic acid bacteria cultures in fermented peach juice was also measured during four weeks of cold storage at 4°C.

Results

Lactobacillus delbrueckii grew well in peach juice, reached nearly 10 × 109 CFU/mL, after 48 hours of fermentation at 30 °C and was capable of more sugar consumption, pH inclination and production of lactic acid during fermentation. After four weeks of cold storage at 4 °C, the viable cell counts of L. delbrueckii were 1.72 × 107 CFU/mL, in fermented peach juice. Lactobacillus casei could not survive in fermented fruit juice after the cold storage.

Conclusions

In conclusion, L. delbrueckii in peach juice was appropriate to produce a probiotic beverage. Therefore, this juice can serve as a healthy beverage for vegetarians and lactose-allergic consumers.

Background

Probiotics have been used for dairy products such as yogurt and yogurt drinks, however cholesterol content and lactose intolerance are important drawbacks. Recently, consumption of non-dairy probiotic food especially for probiotic drink products has been intensified.

Objectives

This research was conducted to determine the suitability of peach as a raw material for producing probiotic peach juice by lactic acid bacteria.

Materials and Methods

Peach juice was inoculated with a 24-hour-old lactic acid bacteria culture and incubated at 30°C. Changes in the pH, titratable acidity, sugar content, and viable cell counts during fermentation under controlled conditions were monitored. Viability of lactic acid bacteria cultures in fermented peach juice was also measured during four weeks of cold storage at 4°C.

Results

Lactobacillus delbrueckii grew well in peach juice, reached nearly 10 × 109 CFU/mL, after 48 hours of fermentation at 30 °C and was capable of more sugar consumption, pH inclination and production of lactic acid during fermentation. After four weeks of cold storage at 4 °C, the viable cell counts of L. delbrueckii were 1.72 × 107 CFU/mL, in fermented peach juice. Lactobacillus casei could not survive in fermented fruit juice after the cold storage.

Conclusions

In conclusion, L. delbrueckii in peach juice was appropriate to produce a probiotic beverage. Therefore, this juice can serve as a healthy beverage for vegetarians and lactose-allergic consumers.

Fermentation;Probiotic;Cultures Fermentation;Probiotic;Cultures http://www.Biotech-health.com/index.php?page=article&article_id=24683 Babak Pakbin Babak Pakbin Department of Food Science and Technology, Faculty of Biosystems Engineering, Campus of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran Department of Food Science and Technology, Faculty of Biosystems Engineering, Campus of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran Seyyed Hadi Razavi Seyyed Hadi Razavi Department of Food Science and Technology, Faculty of Biosystems Engineering, Campus of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran Department of Food Science and Technology, Faculty of Biosystems Engineering, Campus of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran Razzagh Mahmoudi Razzagh Mahmoudi Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran; Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran. Tel: +98-9127868571 Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran; Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, IR Iran. Tel: +98-9127868571 Payman Gajarbeygi Payman Gajarbeygi Department of Food Health and Safety, School of Public Health, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Food Health and Safety, School of Public Health, Qazvin University of Medical Sciences, Qazvin, IR Iran
en 10.17795/bhs-26375 Novel Real Time Polymerase Chain Reaction Approach for Rapid Detection of the Residual Escherichia coli Genomic DNA in Biopharmaceutical Products Establishment of Real Time Polymerase Chain Reaction to Detect Residual gDNA Novel Real Time Polymerase Chain Reaction Approach for Rapid Detection of the Residual <italic>Escherichia coli</italic> Genomic DNA in Biopharmaceutical Products Establishment of Real Time Polymerase Chain Reaction to Detect Residual gDNA research-article research-article Background

Contamination of therapeutic recombinant proteins with residual host cell DNA must be controlled under the regulatory standards.

Objectives

The current study established a new rapid, sensitive real time polymerase chain reaction (PCR) approach to measure the reliably of the residual Escherichia coli (E. coli) host cell genomic DNA in the recombinant streptokinase and alfa interferon preparations.

Materials and Methods

In this assay, a specific primer pair was utilized to amplify a 115 base pair sequence inside the E. coli 16S rRNA using SYBR Green Chemistry. This method enabled the authors to detect a very small quantity of the residual genomic DNA, as low as 0.8 pg, in the protein-based drugs. This method can, therefore, offer a dependable way to quantitatively analyze the major contaminant of biopharmaceutical products, the host cell DNA, during the manufacturing process.

Results

SYBR Green PCR master mix may contain a source of DNA contamination during its manufacturing process.

Conclusions

The current study data showed that E. coli host cell DNA contamination in streptokinase and alfa interferon manufactured in the Pasteur institute of Iran is much lower than the safety limits suggested by the FDA.

Background

Contamination of therapeutic recombinant proteins with residual host cell DNA must be controlled under the regulatory standards.

Objectives

The current study established a new rapid, sensitive real time polymerase chain reaction (PCR) approach to measure the reliably of the residual Escherichia coli (E. coli) host cell genomic DNA in the recombinant streptokinase and alfa interferon preparations.

Materials and Methods

In this assay, a specific primer pair was utilized to amplify a 115 base pair sequence inside the E. coli 16S rRNA using SYBR Green Chemistry. This method enabled the authors to detect a very small quantity of the residual genomic DNA, as low as 0.8 pg, in the protein-based drugs. This method can, therefore, offer a dependable way to quantitatively analyze the major contaminant of biopharmaceutical products, the host cell DNA, during the manufacturing process.

Results

SYBR Green PCR master mix may contain a source of DNA contamination during its manufacturing process.

Conclusions

The current study data showed that E. coli host cell DNA contamination in streptokinase and alfa interferon manufactured in the Pasteur institute of Iran is much lower than the safety limits suggested by the FDA.

Streptokinase;Alfa interferon;Real Time PCR Streptokinase;Alfa interferon;Real Time PCR http://www.Biotech-health.com/index.php?page=article&article_id=26375 Taghi Naserpour Farivar Taghi Naserpour Farivar Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran Babak Mamnoon Babak Mamnoon Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Mohsen Karimi Arzenani Mohsen Karimi Arzenani Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, IR Iran Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, IR Iran Dariush Ilghari Dariush Ilghari Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran. Fax: +98-2833331007 Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Clinical Biochemistry and Genetics, Qazvin University of Medical Sciences, Qazvin, IR Iran. Fax: +98-2833331007
en 10.17795/bhs-25836 Prevalence of Helicobacter pylori in Formaldehyde Fixed Paraffin Embedded Gastric Tissues of Gastric Cancer Patients by Scorpion Real-Time PCR Assay Prevalence of <italic>Helicobacter pylori</italic> in Formaldehyde Fixed Paraffin Embedded Gastric Tissues of Gastric Cancer Patients by Scorpion Real-Time PCR Assay research-article research-article Background

Gastric cancer is the second leading cause of cancer-related deaths worldwide and it seems that environmental and lifestyle factors and infection with Helicobacter pylori (H. pylori) have a major role in the etiology of gastric cancer.

Objectives

The aim of this study was to investigate the presence of H. pylori DNA in archival gastric tissues of patients with gastric cancer disease by rapid, sensitive and specific technique of Scorpion real-time PCR.

Patients and Methods

This retrospective cross-sectional study was performed during the year 2009, on 285 paraffin embedded gastric specimens of patients, who were pathologically proved to have gastric cancer and were admitted to Bou-Ali, Shahid Rajaie and Dehkhoda Hospitals and Bahar and Farzam Private Laboratory of Qazvin city, Iran.

Results

The results of the Scorpion real-time PCR showed that H. pylori DNA was present in 8.42% of the total specimens. Modified McMullen’s staining of paraffin embedded sections were positive in ten patients. There was no significant relationship between the presence of H. pylori, sex, age and place of residence.

Conclusions

Although the existence of H. pylori in gastric tissue samples of patients with gastric cancer is controversial however, our results showed that in our studied specimens a significant number of patients with gastric cancer had H. pylori colonization.

Background

Gastric cancer is the second leading cause of cancer-related deaths worldwide and it seems that environmental and lifestyle factors and infection with Helicobacter pylori (H. pylori) have a major role in the etiology of gastric cancer.

Objectives

The aim of this study was to investigate the presence of H. pylori DNA in archival gastric tissues of patients with gastric cancer disease by rapid, sensitive and specific technique of Scorpion real-time PCR.

Patients and Methods

This retrospective cross-sectional study was performed during the year 2009, on 285 paraffin embedded gastric specimens of patients, who were pathologically proved to have gastric cancer and were admitted to Bou-Ali, Shahid Rajaie and Dehkhoda Hospitals and Bahar and Farzam Private Laboratory of Qazvin city, Iran.

Results

The results of the Scorpion real-time PCR showed that H. pylori DNA was present in 8.42% of the total specimens. Modified McMullen’s staining of paraffin embedded sections were positive in ten patients. There was no significant relationship between the presence of H. pylori, sex, age and place of residence.

Conclusions

Although the existence of H. pylori in gastric tissue samples of patients with gastric cancer is controversial however, our results showed that in our studied specimens a significant number of patients with gastric cancer had H. pylori colonization.

Scorpion;Real-Time PCR;Helicobacter pylori;Cancer Scorpion;Real-Time PCR;Helicobacter pylori;Cancer http://www.Biotech-health.com/index.php?page=article&article_id=25836 Amir Farzam Amir Farzam Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Pathology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Pathology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Reza Najafipour Reza Najafipour Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Genetics, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Genetics, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Pouran Johari Pouran Johari Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Taghi Naserpour Farivar Taghi Naserpour Farivar Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-2813338034 Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Microbiology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cell and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-2813338034
en 10.17795/bhs-26379 The Role of Calcium in Calprotectin Dimerization as a Cancer Biomarker The Role of Calcium in Calprotectin Dimerization as a Cancer Biomarker research-article research-article Conclusions

The expression of recombinant calprotectin, as an effective protein, can help in diagnosis or treatment of inflammatory and cancer processes in the future. Furthermore, Ca2+ induced a partial change in secondary and tertiary structure of calprotectin subunits and this change is probably necessary for protein dimerization.

Results

Interaction of S100A8 and S100A9 in the presence of Ca2+ were revealed by decreasing the emission intensity of intrinsic fluorescence and increasing of the external fluorescence and also changes in the CD spectra of subunits after Ca2+ interactions.

Objectives

The aim of this study was to investigate the effects of calcium in calprotectin dimerization as a cancer biomarker.

Materials and Methods

Heterodimeric calprotectin was formed with incubation of recombinant S100A8 and S100A9 subunits in the presence of Ca (1 mM), at 25˚C for 15 minutes. Tertiary and secondary structures of S100A8, S100A9 and their complex were investigated, using fluorescence and circular dichroism (CD) spectroscopy, respectively.

Background

S100A8 and S100A9 as two subunits of heterodimeric calprotectin are identified mainly in leukocytes and are involved in inflammatory processes and several cancerous pathogens. This study was performed in order to evaluate the interaction of recombinant calprotectin subunits and to estimate calprotectin’s tertiary and secondary structures.

Conclusions

The expression of recombinant calprotectin, as an effective protein, can help in diagnosis or treatment of inflammatory and cancer processes in the future. Furthermore, Ca2+ induced a partial change in secondary and tertiary structure of calprotectin subunits and this change is probably necessary for protein dimerization.

Results

Interaction of S100A8 and S100A9 in the presence of Ca2+ were revealed by decreasing the emission intensity of intrinsic fluorescence and increasing of the external fluorescence and also changes in the CD spectra of subunits after Ca2+ interactions.

Objectives

The aim of this study was to investigate the effects of calcium in calprotectin dimerization as a cancer biomarker.

Materials and Methods

Heterodimeric calprotectin was formed with incubation of recombinant S100A8 and S100A9 subunits in the presence of Ca (1 mM), at 25˚C for 15 minutes. Tertiary and secondary structures of S100A8, S100A9 and their complex were investigated, using fluorescence and circular dichroism (CD) spectroscopy, respectively.

Background

S100A8 and S100A9 as two subunits of heterodimeric calprotectin are identified mainly in leukocytes and are involved in inflammatory processes and several cancerous pathogens. This study was performed in order to evaluate the interaction of recombinant calprotectin subunits and to estimate calprotectin’s tertiary and secondary structures.

Calcium;Calprotectin;Biomarkers;Cancer;Fluorescence Spectroscopy Calcium;Calprotectin;Biomarkers;Cancer;Fluorescence Spectroscopy http://www.Biotech-health.com/index.php?page=article&article_id=26379 Fatemeh Nemati Nikoo Fatemeh Nemati Nikoo Department of Nanotechnology, Pharmacuitical Sciences Branch, Islamic Azad University, Tehran, IR Iran Department of Nanotechnology, Pharmacuitical Sciences Branch, Islamic Azad University, Tehran, IR Iran Koorosh Goodarzvand Chegini Koorosh Goodarzvand Chegini Department of Biochemistry and Genetic, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Biochemistry and Genetic, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Reza Najafi Pour Reza Najafi Pour Department of Biochemistry and Genetic, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Biochemistry and Genetic, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Nematollah Gheibi Nematollah Gheibi Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-2813338034 Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran; Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran. Tel: +98-2813338034
en 10.17795/bhs-26373 Inhibitory Effects of a Palladium Complex on the Activity, Stability, and Structure of Tyrosinase Enzyme Inhibitory Effects of a Palladium Complex on the Activity, Stability, and Structure of Tyrosinase Enzyme research-article research-article Background

Tyrosinase, as a copper-containing enzyme, is widely distributed in different levels of life span. It is also a key enzyme in melanin biosynthesis, which plays a crucial role in determining the color of mammalian skin and hair.

Objectives

The current study aimed to determine the effect of a palladium complex on cresolase and catecholase reactions of mushroom tyrosinase (MT).

Materials and Methods

The MT kinetics parameters were obtained from double reciprocal plots of Lineweaver-Burk and the inhibition constants (Ki) were determined by the secondary plots. Thermodynamic parameters were obtained from thermal and chemical denaturation of the tyrosinase with and without the presence of palladium complex. The tertiary and secondary structures of tyrosinase were detected by fluorescent and Circular Dichroism (CD) techniques.

Results

The inhibition modes of palladium complex were competitive in both activities of the enzyme with Ki values of 3.74 and 10.55 μM for cresolase and catecholase activities, respectively. In thermal denaturation, the melting points (Tm) of the enzyme were 59.4˚C and 51˚C for the sole enzyme and its treatment by palladium, respectively. In chemical denaturation, the magnitudes of half denaturant concentration (Cm) were 1 μM vs. 1.36μM and the free energy of Gibss (ΔGH2O) were calculated 9.3 vs. 7.5 kJ/M for the sole enzyme and its treatment by palladium, respectively.

Conclusions

In overall the palladium complex acted as a good inhibitor of tyrosinase and induced the enzyme thermodynamic and conformational instability, therefore it can be considered in the hyper expression of tyrosinase in melanoma cancer.

Background

Tyrosinase, as a copper-containing enzyme, is widely distributed in different levels of life span. It is also a key enzyme in melanin biosynthesis, which plays a crucial role in determining the color of mammalian skin and hair.

Objectives

The current study aimed to determine the effect of a palladium complex on cresolase and catecholase reactions of mushroom tyrosinase (MT).

Materials and Methods

The MT kinetics parameters were obtained from double reciprocal plots of Lineweaver-Burk and the inhibition constants (Ki) were determined by the secondary plots. Thermodynamic parameters were obtained from thermal and chemical denaturation of the tyrosinase with and without the presence of palladium complex. The tertiary and secondary structures of tyrosinase were detected by fluorescent and Circular Dichroism (CD) techniques.

Results

The inhibition modes of palladium complex were competitive in both activities of the enzyme with Ki values of 3.74 and 10.55 μM for cresolase and catecholase activities, respectively. In thermal denaturation, the melting points (Tm) of the enzyme were 59.4˚C and 51˚C for the sole enzyme and its treatment by palladium, respectively. In chemical denaturation, the magnitudes of half denaturant concentration (Cm) were 1 μM vs. 1.36μM and the free energy of Gibss (ΔGH2O) were calculated 9.3 vs. 7.5 kJ/M for the sole enzyme and its treatment by palladium, respectively.

Conclusions

In overall the palladium complex acted as a good inhibitor of tyrosinase and induced the enzyme thermodynamic and conformational instability, therefore it can be considered in the hyper expression of tyrosinase in melanoma cancer.

Mushroom;Tyrosinase;Palladium;Inhibition Mushroom;Tyrosinase;Palladium;Inhibition http://www.Biotech-health.com/index.php?page=article&article_id=26373 Nematollah Gheibi Nematollah Gheibi Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IR Iran Nasibe Yaghouby Nejad Nasibe Yaghouby Nejad Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, IR Iran Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, IR Iran Mehdi Sahmani Mehdi Sahmani Department of Biochemistry, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Biochemistry, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran Department of Biochemistry, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran; Department of Biochemistry, School of Medicine, Qazvin University of Medical Sciences, Qazvin, IR Iran