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23407587
Development in Immunoprophylaxis against Rabies for Animals and Humans
Rabies is a fatal neurological disease and a persistent global problem. It is spread primarily by domestic dogs but other canid, viverrid (skunks and raccoons) and chiropteran species are considered as the most efficient vectors of the disease. Since dogs are the main perpetuator of rabies, special attention has to be given to bring all the dogs including unauthorized stray dogs under immunization umbrella in order to control rabies. Vaccination is the only way to combat the disease before and after exposure or infection as there is no treatment available once the symptoms have appeared. After the first crude nerve tissue vaccine developed by Pasteur in 1885, a number of rabies vaccines for animal and human use have been developed with varying degree of safety and efficacy over the years. Presently, cell culture based inactivated rabies vaccines are largely used in most of the parts of the world. However, these vaccines are too expensive and unaffordable for vaccination of people and animals in developing countries. The comparatively cheaper inactivated nerve tissues vaccines can cause serious side-effects such as autoimmune encephalomyelitis in inoculated animals and production has been discontinued in several countries. Although attenuated live vaccines can efficiently elicit a protective immune response with a smaller amount of virus, they sometimes can cause rabies in the inoculated animals by its residual virulence or pathogenic mutation during viral propagation in the body. New-generation rabies vaccines generated by gene manipulation although in experimental stage may be a suitable alternative to overcome the disadvantages of the live attenuated vaccines. So, awareness must be created in general public about the disease and the cell culture based vaccines available in the market should be recommended for wide scale use to prevent and control this emerging and reemerging infectious disease in foreseeable future.
Lyssavirus, Rabies virus, Rabies, Vaccination, Zoonoses
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https://www.ajmb.org/En/Article.aspx?id=29
https://www.ajmb.org/PDF/En/FullText/29.pdf
SukdebNandiVirology Laboratory, Center for Animal Disease Research and Diagnosis (CADRAD), Indian Veterinary Research Institute (IVRI) , Izatnagar, U.P, India 95
ManojKumarVirology Laboratory, Center for Animal Disease Research and Diagnosis (CADRAD), Indian Veterinary Research Institute (IVRI) , Izatnagar, U.P, India 100
en
23407454
The Effect of Human Chorionic Gonadotropin Treatment on Recipient Mouse Germ Cell Proliferation Following Spermatogonial Stem Cell Transplantation of Neonatal Donor Mice
Spermatogonia are the male germ line stem cells whose life long expansion is needed for permanent production of spermatozoa. The present study was designed to examine the effect of hCG treatment on germ cell proliferation following stem cell transplantation in mice. Spermatogonial stem cells were isolated from neonatal mice testes and characterized by alkaline phosphatase, immunoreactivity and morphological analysis. hCG was injected into normal and cell transplanted mice. We then evaluated the testosterone levels and cell number in normal mice. After that, cyclin B1 gene expression was investigated in transplanted mice. Different doses of busulfan were injected to investigate the effects of chemotherapy on morphological criteria and preparation of recipient mice for transplantation. In this report we show proliferative potential of spermatogonial stem cells after cytotoxic treatment, transplantation efficiency by semi-quantitative RT-PCR, and hCG effect on stem cell regeneration in normal mice and following cell transplantation. The results indicate that spermatogonial stem cells can proliferate after transplantation, and the efficiency of their transplantation depends on hormonal treatment. Therefore, hormonal treatment after stem cell transplantation will be a powerful avenue for increasing the efficiency of transplantation and fertility restoration.
Busulfan, Cell proliferation, hCG, Transplantation
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35
https://www.ajmb.org/En/Article.aspx?id=27
https://www.ajmb.org/PDF/En/FullText/27.pdf
Mohammad MehdiAkhondiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran13
RezaAkbarzadeh NajarReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran94
MahmoodJeddi-TehraniMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR , Tehran, Iran15
Mohammad RezaSadeghiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran , Iran40
Amir-HassanZarnaniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran7
HodjattallahRabbaniImmune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska Institute , Stockholm, Sweden24
SheidaSalehkhouReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran96
LeilaEiniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran97
FatemehHoseinzadehReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran98
MahnazHeidariReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran99
en
23408735
Production and Characterization of Mouse Monoclonal Antibodies Recognizing Multiple Subclasses of Human IgG
Different IgG subclass profiles are produced in response to different antigenic stimuli in a variety of diseases. IgG subclass levels may reflect disease severity. Quantification of IgG subclasses depends on the availability of specific Monoclonal antibodies (MAbs). In the present study seven hybridoma clones producing MAbs reactive with multiple subclasses of human IgG were established. Splenocytes from Balb/c mice immunized with Fc fractions of human IgG1 or IgG2 myeloma proteins were fused with mouse myeloma cells. Fused cells were selected and cloned by limiting dilution assay. Antibody secreting cells were screened by Enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAbs was further analyzed, using a panel of purified human myeloma paraproteins of different IgG subclasses by ELISA and immunoblotting. Cross-reactivity to immunoglobulins (Igs) of other species was studied by indirect ELISA using serum samples collected from 9 animals. The MAbs were found to react with triple IgG subclasses, including IgG1,2,4 (n=4) and IgG1,2,3 (n=3). Immunoblotting studies revealed recognition of linear (n=4) or conformational (n=3) epitopes by these MAbs. The most abundant cross-reactivity (71.4%) was observed with monkey Ig while no cross-reactivity was detected with hen and cat sera. The MAbs mostly displayed a restricted pattern of cross-reactivity and one of them did not bind to any of the animal sera tested. The affinity constant of 3 MAbs was measured by ELISA. Based on the data obtained from this study, mouse MAbs reactive with multiple human IgG subclasses are directed to a variety of immunogenic epitopes, mostly shared with IgG of other species. These MAbs are valuable tools for purification of non-reactive IgG subclasses through negative affinity chromatography. These MAbs could also provide an opportunity for epitope mapping of the Fc region of IgG, as well as serological phylogenetic studies.
ELISA, Hybridoma, IgG, Isotype, Monoclonal antibody, Subclass
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https://www.ajmb.org/En/Article.aspx?id=28
https://www.ajmb.org/PDF/En/FullText/28.pdf
FatemehHajighasemiDepartment of Immunology, School of Medicine, Shahed University , Tehran, Iran4
FazelShokriMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR , Tehran, Iran5
en
23407146
Intermittent Feeding Attenuates Clinical Course of Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice
Multiple Sclerosis (MS) is an autoimmune inflammatory, demyelinating disease of human central nervous system. Experimental Autoimmune Encephalomyelitis (EAE) is the commonly used animal model of MS. Calorie restriction has been found to reduce inflammation and autoimmune responses and promote neuroprotection. In this study we evaluated the effects of intermittent feeding protocol of the calorie restriction in a mouse model of EAE. Fifty four female mice (C57BL/6) were used in this study. The animals were divided into two dietary groups: ad libitum (AL) (n=29) with free access to food and water and intermittent feeding (IF) (n=25) with access to food on alternate days. After 8 weeks, EAE was induced in animals by immunization with MOG antigen (Hooke labs, Lawrence, MA, USA) subcutaneously. AL and IF groups were then further divided into two groups each: AA (ad libitum until the end of study) (n=16) and AI (subjected to intermittent feeding regimen after immunization day) (n=13). The IF group was divided into II (continued intermittent feeding regimen until the end of study) (n=13) and IA (changed to AL regimen after immunization day) (n=12). All the animals were behaviorally monitored for 35 days after immunization and observed daily for the signs and severity of disease with EAE scoring scale [0-5] and cumulative disease index (CDI) score. Intermittent feeding significantly reduced the incidence of EAE in IF groups (AI 0%, II 18.5%, IA 22.2%, p<0.05). In addition, intermittent feeding significantly delayed the onset of EAE in AI group (p<0.05) and also, intermittent feeding significantly reduced the severity of disease in II and IA groups (AA vs. II, p<0.05 & AA vs. IA p<0.05) groups. The CDI was also significantly reduced in intermittent feeding fed groups [AI, II and IA compared to AA group (P<0.05, <0.01, <0.05 respectively)]. Intermittent feeding regimen protocol of the calorie restriction significantly suppressed EAE incidence, induction, and severity. The results of this study suggest possible role of intermittent feeding in the treatment of Multiple Sclerosis patients.
Experimental autoimmune encephalomyelitis, Intermittent feeding, Mice, Multiple sclerosis
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https://www.ajmb.org/En/Article.aspx?id=30
https://www.ajmb.org/PDF/En/FullText/30.pdf
LayaKafamiDepartment of Pathobiology, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran101
MohsinRazaApplied Neuroscience Research Center, Baqiyatallah University of Medical Sciences , Tehran, Iran102
AlirezaRazaviDepartment of Pathobiology, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran103
AbbasMirshafieyDepartment of Pathobiology, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran104
MansoorehMovahedianDepartment of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University , Tehran, Iran105
Mohammad RezaKhorramizadehDepartment of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences , Tehran, Iran61
en
23409223
Editorial
New advances in Genetic Technologies are changing the lives of millions of people around the world in important ways. These advances have also raised difficult ethical and legal questions for policy makers in many countries. Therefore, there is a great interest and need by experts in both governmental and non-governmental institutions to have a deeper understanding of the issues involved for establishing the necessary societal rules to regulate the use of genetic technologies in the fields of Medicine, Veterinary Science and Agriculture. To address these issues and provide a forum for a scientific discussion at a national level, the Avicenna Research Institute is planning to hold a conference in November 2010, entitled 'Genetics: Law, Ethics and Psychology'. The conference will particularly focus on the use of new genetic technologies and its impacts on the society from the legal and ethical point of view. In view of the fact that better understanding of the genetic basis of human behavior and physiology is imperative to comprehend the more complex topics of the conference, the genetics of human behavior will also be discussed in the convention.
The conference will aim to address the specific areas of concern in the use of genetic technologies in human health as follows:
• Assisted Reproduction Techniques (e.g. in-vitro fertilization)- ART are used to help fertility problems. Ethical issues are around the creation, selection, and disposal of embryos. These technologies can also require the use of sperm, eggs, or wombs from other women who are unrelated to the real parents and are not expected to play a role in raising the child.
• Pre-implantation Genetic Diagnosis – In this technique an embryo at 6-10 cell stage can be tested and selected for or against a specific sex, disease and physical condition. Although, this method can be used for treatment purposes, however the ethical issues are around the use of this technology by parents to select a specific baby based on personal desires.
• Cloning and Stem cells - Cloning is an essential tool of modern biology which has led to important drugs and new therapies. Cloning also has helped the understanding of genetic basis of human development and disease. Cloning has a potential to be used in producing a lifetime supply of therapeutic stem cells that are genetically matched to a patient. The ethical issues around cloning concern the production and destruction of a two-to-four-day-old embryo to make a line of embryonic stem cells. Another, concern is assuring that women donating eggs for research give proper informed consent. Some fear that a cloned embryo could be implanted into a woman resulting in a baby; a cloned human being.
• Animal and Plant Cloning – Cloning animals and plants for specific purposes are becoming possible with new genetic technologies. Specific recombinant proteins to be used as therapeutic drugs are being produced in some animals and specific plants with certain characteristics are now possible to be made and some are currently available in the market for human use. The ethical issues around the animal and plant cloning are the consequence of releasing such new species in the environment and its impact on the society and human health.
• Biobank – Biospecimens are being stored in public and private repositories and contain genetic material to identify gene variations associated with human diseases and lead to diagnostic tests and targeted treatments for specific diseases. Ethical issues are around the methods used in obtaining informed consent, protect privacy and disclose of research results, ownership of biospecimens as intellectual property and the ethical use of the biospecimens.
• New frontiers in genetics research–There have been many new genetic technologies developed and there are plans to be applied in human health.
• The research areas such as Human Behavior, Epigenetics, Nutrigenomics are some of the topics that are planned to be addressed in the forum.
I look forward to your active participation in the conference and receive articles examining the issues related to ethical and legal aspects of Medical Biotechnology.
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https://www.ajmb.org/En/Article.aspx?id=157
https://www.ajmb.org/PDF/En/FullText/157.pdf
AliM. ArdekaniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran2
en
23407328
Flow Cytometric Analysis of 4-HPR-induced Apoptosis and Cell Cycle Arrest in Acute Myelocytic Leukemia Cell Line (NB-4)
In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or programmed cell death (apoptosis) can be induced by variety of agents including: Vitamin analogs, demethylating agents, cyclic AMP analogs and anti-proliferative agents. To the best of our knowledge there has been not any study specifically to analyze apoptotic and anti-proliferative effects of 4-HPR (a vitamin analog) in NB-4 cell line. To test whether this drug has activity in acute myeloid leukemia (AML), we first analyzed the anti-proliferative effect of 4-HPR in one AML cell line (NB-4) using MTT Assay. Next we tested whether this drug induced apoptotic cell death. The ability of this compound to induce apoptosis of cancer cells was examined by Annexin V-FITC Assay using Flow cytometry. We also analyzed the cell cycle progression by PI staining using flow cytometry. Using MTT assay, NB-4 cells exhibited increased inhibition of proliferation at micromolar concentrations of 4-HPR at 24, 48 and 72 hrs post treatment. Flow cytometry analysis indicates that 4-HPR is a potent inducer of in vitro apoptotic cell death, and cell cycle analysis revealed an increase in S phase population. In total, the results indicate that 4-HPR is a strong inhibitor of AML cell proliferation and a potent inducer of in vitro apoptotic cell death. Further studies are required to evaluate the in vitro effects of 4-HPR in AML blasts derived from AML patients.
Acute myelocytic leukemia, Apoptosis, Cell differentiation, Flow cytometry, 4-HPR
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https://www.ajmb.org/En/Article.aspx?id=26
https://www.ajmb.org/PDF/En/FullText/26.pdf
ShahrzadSoleymani FardBiology Department, Faculty of Basic Science, Science and Research Branch, Islamic Azad University , Tehran, Iran92
MahmoodJeddi-TehraniMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR , Tehran, Iran15
Mohammad MehdiAkhondiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , Tehran, Iran13
MehrdadHashemiMolecular Genetic Department, Tehran Medical Branch, Islamic Azad University , Tehran, Iran93
AliM. ArdekaniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, , Tehran,, Iran2
en
23408769
Development of a Latex Agglutination Test as a Simple and Rapid Method for Diagnosis of Trichomonas vaginalis Infection
Trichomoniasis is a worldwide infection and due to its complications rapid and accurate diagnosis of infection especially in pregnant women is very important. In this study, development of a latex agglutination test using native antigens for rapid diagnosis of trichomoniasis is investigated. Trichomonas vaginalis was harvested from TYIS33 culture medium and anti Trichomonas vaginalis antiserum was raised in rabbits. Salt precipitation method was used for antibody purification. Polyesteren latex particles coated with purified antibody and used for detection of Trichomonas vaginalis. Clinical samples of vaginal discharge were collected from 500 women and examined for Trichomonas vaginalis by using wet mount, culture and latex agglutination tests. Sensitivity and specificity of latex test was determined considering culture as golden standard. Sensitivity and specificity of latex agglutination test was 100% and 81% and those of wet mount were 33.3% and 100%, respectively. Positive and negative predictive values of latex agglutination test were 6% and 100%, respectively. Due to inconvenient sensitivity and specificity of the latex agglutination test developed in this study, further work is recommended to improve the test.
Latex agglutination test, Sensitivity, Specificity, Trichomonas vaginalis
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https://www.ajmb.org/En/Article.aspx?id=31
https://www.ajmb.org/PDF/En/FullText/31.pdf
HosseinYousofi DaraniDepartment of Parasitology, Cell and Molecular Research Center, Faculty of Medicine, Shahrekord University of Medical Sciences , Shahrekord, Iran106
FiruzehAhmadiDepartment of Parasitology, Shahrekord University of Medical Sciences , Shahrekord, Iran107
NozhatZabardastDepartment of Parasitology, Shahrekord University of Medical Sciences , Shahrekord, Iran108
Hossein AliYousefiDepartment of Parasitology, Isfahan University of Medical Sciences , Isfahan, Iran109
HedayatShirzadDepartment of Immunology, Faculty of Medicine, Shahrekord University of Medical Sciences , Shahrekord, Iran110