Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 21
Filter
1.
J Vector Borne Dis ; 60(4): 444-448, 2023.
Article in English | MEDLINE | ID: mdl-38174525

ABSTRACT

Background & objectives: The resistance to insecticide among Anopheles stephensi population due to insecticide selection pressure has been previously reported from Iran. The current study was performed to evaluate the susceptibility of different insecticide reagents against An. stephensi by bioassay and molecular methods in Saravan County, a malaria-endemic area in southeastern Iran. Methods: An. stephensi mosquitoes were collected from different larval habitats in Saravan City, southeastern Iran in 2022. At first, the susceptibility of collected samples for DDT, permethrin, and deltamethrin were evaluated by bioassay test. The collected mosquitoes were then evaluated for the presence of different kdr mutations. Results: Insecticide susceptibility tests were conducted on the field population of An. stephensi from Saravan, revealing its potential resistance to pyrethroids and DDT. Of the 150 An. stephensi samples, 4 % carried the kdr L1014F mutation as heterozygous and the rest of them were homozygous L1014 wild type. Interpretation & conclusion: The current study revealed the presence of L1014F mutation for the first time in Iran. So, further monitoring of kdr mutations in the VGSC gene and resistance phenotypes should be performed.


Subject(s)
Anopheles , Insecticides , Pyrethrins , Animals , Insecticides/pharmacology , Anopheles/genetics , DDT , Insecticide Resistance/genetics , Iran , Pyrethrins/pharmacology , Mutation
2.
Avicenna J Med Biotechnol ; 9(3): 150-154, 2017.
Article in English | MEDLINE | ID: mdl-28706612

ABSTRACT

BACKGROUND: Recurrent Spontaneous Abortion (RSA) is caused by multiple genetic and non-genetic factors. Around 50% of the RSA cases have no known etiology and are considered as Unexplained RSA (URSA). Estrogens, via binding to their receptors, play an important role in female reproduction. This study aimed to investigate whether single nucleotide polymorphisms (SNPs; +1082G/A, +1730G/A and rs1256030 C/T) in the estrogen receptor beta (ESR2) gene are associated with susceptibility to URSA in a population of Iranian women. METHODS: In this case-control study, the study groups consisted of 240 subjects with a history of URSA and 102 fertile women as controls. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were measured on day 2-3 of menstrual cycle. Two functional SNPs, +1082G/A (a silent mutation in exon 5) and +1730G/A (3' untranslated region of the exon 8), and one intron, rs1256030C/T, in the ESR2 gene were genotyped, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: Serum levels of LH were significantly increased in URSA women. No significant differences in distribution of +1082G/A, +1730G/A and rs1256030C/T between URSA and control groups were observed. CONCLUSION: Our findings suggest that the studied SNPs on ESR2 gene may not be associated with URSA.

3.
Hepat Mon ; 16(9): e39416, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27822265

ABSTRACT

BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are major health problem in the world. Hairdressers (barbers) are in continuous contact with scissors and blades, and are considered a high-risk group for these infections. OBJECTIVES: The aim of this study was to analyze the prevalence of hepatitis B and C infections in barbers in Tehran and to evaluate their attitudes and knowledge about the occupational risk of these infections. METHODS: Six hundred eleven barbers were included in this study. A group of 556 bakers were also selected from the same regions, as a low-risk control group. Serum levels of hepatitis B surface antigen (HBsAg), HBsAg-specific antibody (HBsAb), hepatitis B core antigen-specific antibody (HBcAb), and hepatitis C virus-specific (anti-HCV) antibody markers were measured with the enzyme-linked immunosorbent assay (ELISA). Participants were interviewed using a questionnaire consisting of four sections: demographic information, awareness, behavior, and personal attitudes. RESULTS: There were no significant differences in the frequency of HBsAg between the two groups. However, the frequency of HCV Ab in barbers was significantly higher than that in bakers (P < 0.005). In addition, the frequency of HBsAb marker in barbers was significantly correlated with increased awareness (P < 0.05) and number of tattoos (P < 0.001). HBcAb marker was significantly correlated with age (P < 0.001) and duration of professional career (P < 0.005). With age, barbers' attitudes improved significantly (P < 0.05). CONCLUSIONS: Being a barber alone is not a potential risk factor for HBV infection, while HCV infection is still an occupational health hazards for barbers. We suggest more extensive case-control studies with regard to rates of hepatitis B and C markers among barbers in other Iranian cities to assess the incidence of hepatitis B and C infections among this population.

4.
Infect Dis (Lond) ; 48(2): 127-32, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26439274

ABSTRACT

BACKGROUND: Filamentous hemagglutinin (FHA) is a principal virulence factor, an important immunogenic antigen of Bordetella pertussis, and a major component of many acellular pertussis vaccines. In the present study, the human antibody response to different regions of FHA was determined in healthy children and adults vaccinated with either whole-cell or acellular pertussis vaccines. METHODS: To define the immunodominant regions of FHA, four overlapping recombinant fragments were expressed and produced in Escherichia coli and then purified by His-tagged based affinity chromatography. Two groups comprising healthy preschool children (n = 50) and adults (n = 26) were vaccinated with a single dose of commercial whole-cell and acellular DTaP vaccines, respectively. An antigen-based ELISA was applied to measure serum levels of anti-FHA antibody to both native and recombinant proteins in vaccinated volunteers. RESULTS: In both groups of vaccinated individuals, the anti-FHA antibody response was mainly directed against epitopes located within a fragment of FHA spanning amino acid residues 1877-2250 of the mature FHA molecule (p < 0.001). No or little antibody was detected against the other recombinant segments of FHA. CONCLUSION: Our results suggest that the human antibody response to FHA is directed to an immunodominant region located within residues 1877-2250 of the FHA molecule. Characterization and epitope mapping of the major components of acellular pertussis vaccine and future modifications in vaccine formulation may improve its efficacy and protectivity.


Subject(s)
Adhesins, Bacterial/immunology , Antibodies, Bacterial/blood , Pertussis Vaccine/administration & dosage , Pertussis Vaccine/immunology , Virulence Factors, Bordetella/immunology , Adult , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Epitopes, B-Lymphocyte/immunology , Female , Healthy Volunteers , Humans , Immunodominant Epitopes/immunology , Male , Vaccines, Acellular/administration & dosage , Vaccines, Acellular/immunology , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/immunology
5.
J Reprod Immunol ; 110: 48-53, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25989617

ABSTRACT

Unexplained recurrent spontaneous abortion (URSA) has been suggested to be associated with the failure of fetal-maternal immunological tolerance in which the regulatory T lymphocytes (Tregs) play a crucial role. This study evaluated the association between single-nucleotide polymorphisms (SNPs) in the forkhead/winged helix transcription factor (FOXP3) gene, a key factor for the development and function of Tregs, and URSA, in an Iranian population. In this case-control study, 195 patients with a history of URSA and101 healthy women were included as case and control groups respectively. Four SNPs in the FOXP3 gene, two in the promoter region: -924A/G and -3279C/A, and two intronic, -20G/A and +459T/C, were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The -924A/G (p<0.0001) and -20G/A (p=0.008) polymorphisms were found to be associated with URSA. The respective odds ratios (OR) for bearing -924A/G and -20G/A gene polymorphisms were 4.1 [95% CI 2.3-7.5] and 2.1 [95% CI 1.2-3.6] fold higher in URSA women than those in controls. Thus, there were significant differences in the distribution of A and G alleles of -924A/G and -20G/A between URSA and controls (p=0.001, OR; 3.6 [95% CI 2.1-6.1] and p=0.006, OR; 1.6 [95% CI 1-2.6] respectively). No associations were found for -3279C/A and +459T/C polymorphisms between URSA and controls. These results suggest that polymorphisms of the FOXP3 gene might confer susceptibility to URSA, probably by altering FOXP3 function and/or its expression.


Subject(s)
Abortion, Habitual/genetics , Forkhead Transcription Factors/genetics , Genetic Predisposition to Disease , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Adult , Case-Control Studies , Female , Humans
6.
Monoclon Antib Immunodiagn Immunother ; 34(1): 36-43, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25723282

ABSTRACT

The gene that codes for the CD44 family members consists of 20 exons, nine of which encode the standard form of the molecule. The other exons can be inserted in various combinations into the membrane proximal region of the extracellular domain of the protein, giving rise to variant isoforms (CD44v). CD44 variants, especially the CD44v6, have been reported to regulate tumor invasion, progression, and metastasis of carcinomas. Producing a high affinity monoclonal antibody against human CD44v6 provides a powerful tool to monitor and trace CD44v6 function in different biological fluids. In this study, a synthetic peptide from CD44v6 was conjugated to keyhole limpet hemocyanin (KLH) and injected into BALB/c mice. Splenocytes from the immunized mice were fused with murine SP2/0 myeloma cells followed by selection of antibody producing hybridoma cells. After screening of hybridoma colonies by ELISA, high affinity antibodies were selected and purified by affinity chromatography. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibodies. Six stable hybridoma cell lines, designated as 1H1, 1H2, 2A12, 2G11, 3H3, and 3H7, were obtained. Flow cytometry and immunocytochemistry results showed that the new monoclonal antibodies recognized CD44v6 on the cell surface. This novel panel of anti-CD44v6 antibodies has the potential for investigating the role of CD44v6 in cancer pathogenesis.


Subject(s)
Antibodies, Monoclonal/biosynthesis , Antigens, Neoplasm/analysis , Hyaluronan Receptors/analysis , Animals , Antibodies, Monoclonal/chemistry , Antibody Specificity , Antigens, Neoplasm/immunology , Blotting, Western , Cell Line , Cell Line, Tumor , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Founder Effect , Haptens/chemistry , Haptens/immunology , Hemocyanins/chemistry , Hemocyanins/immunology , Humans , Hyaluronan Receptors/immunology , Hybridomas/immunology , Immunization , Immunoconjugates/administration & dosage , Immunoconjugates/chemistry , Immunoconjugates/immunology , Immunohistochemistry , Mice , Mice, Inbred BALB C , Peptides/administration & dosage , Peptides/chemical synthesis , Peptides/immunology , Protein Structure, Tertiary
7.
J Reprod Infertil ; 16(4): 193-8, 2015.
Article in English | MEDLINE | ID: mdl-27110516

ABSTRACT

BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) single-nucleotide polymorphisms (SNPs) C677T and A1298C have been described as strong risk factors for idiopathic recurrent miscarriage (RM). However, very few studies have investigated the association of paternal MTHFR SNPs with RM. The aim of the present study was to evaluate the prevalence of paternal C677T and A1298C SNPs among Iranian RM couples. METHODS: The study subjects comprised 225 couples with more than three consecutive pregnancy losses, and 100 control couples with no history of pregnancy complications. All females in the case group had MTHFR polymorphisms; and genotype SNPs were analyzed by PCR-RFLP. Groups were statistically compared using Mann Whitney U-test and Chi-square statistical tests. The p<0.05 were considered significant. RESULTS: Statistically significant difference was detected in the frequency of MTHFR SNPs in male partners of the two groups (p=0.019). Combined heterozygosity of MTHFR polymorphisms was a common phenomenon in the males; 52 (23.1%) and 14 (14%) of males in RM and control groups, respectively. Absence of combined homozygosity for both SNPs in all studied groups/genders was observed. CONCLUSION: The MTHFR gene composition of male partners of RM couples may contribute to increased risk of miscarriage.

8.
Iran J Immunol ; 11(2): 97-104, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24975966

ABSTRACT

BACKGROUND: Pro-inflammatory and anti-inflammatory cytokines and polymorphisms of their genes have been described to be involved in the pathogenesis of recurrent miscarriage (RM). OBJECTIVE: To investigate the association between RM and five polymorphisms of cytokine genes, interleukin 10 (IL-10), (-592 A/C, -819 C/T, -1082 A/G), IL-6 (-174 C/G) and IL-17 (-197 G/A) in Iranian women. METHOD: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to determine the frequencies of the IL-6, IL-10 and IL-17 gene polymorphisms in 85 women with RM compared with 104 healthy controls. RESULTS: The frequencies of IL-10 promoter gene polymorphisms (-592 A/C and -819 C/T) were significantly higher in RM women than those in controls (p=0.003). However, no statistically significant differences were observed in the frequencies of IL-6 (-174 C/G), IL-10 (-1082 A/G) and IL-17 (-197 G/A) polymorphisms between RM women and controls. CONCLUSION: These results suggest that IL-10 gene polymorphism screening might have some relevance in patients with RM, a suggestion which requires further studies.


Subject(s)
Abortion, Habitual/genetics , Interleukin-10/genetics , Interleukin-17/genetics , Interleukin-6/genetics , Polymorphism, Genetic , Adult , Alleles , Case-Control Studies , Female , Gene Frequency , Genetic Association Studies , Genetic Predisposition to Disease , Genotype , Humans , Polymorphism, Single Nucleotide , Pregnancy
9.
Iran J Reprod Med ; 12(6): 395-400, 2014 Jun.
Article in English | MEDLINE | ID: mdl-25071847

ABSTRACT

BACKGROUND: Recurrent pregnancy loss (RPL) is a multifactorial disorder. Environmental factors and genetics can affect pregnancy outcomes. OBJECTIVE: Conflicting data suggest an association between estrogen receptor alpha (ESR1) gene polymorphisms and RPL. In this study, such association was investigated in Iranian women with RPL. MATERIALS AND METHODS: In this case control study, blood samples were collected from 244 women with a history of three or more consecutive pregnancy losses and 104 healthy women with at least two live births. Using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP), we studied -397C/T and -351A/G polymorphisms on ESR1 gene in case and control subjects. RESULTS: The genotypic frequencies of -397C/T and -351A/G polymorphisms on ESR1were not significantly different between RPL and control groups (p=0.20 and p=0.09, respectively). A significantly negative correlation was observed between -397C/T and -351A/G (r=-0.852, p<0.001) in RPL women and complete linkage disequilibrium between the investigated polymorphisms was found (D': 0.959; r-square= 0.758, p<0.001). CONCLUSION: This investigation suggests that the analyzed polymorphisms on ESR1gene are not associated with an increased risk of RPL in the studied population.

10.
Cancer Cell Int ; 14: 54, 2014.
Article in English | MEDLINE | ID: mdl-24966802

ABSTRACT

BACKGROUND: Many types of tumors are organized in a hierarchy of heterogeneous cell populations with different molecular signature. Such heterogeneity may be associated with different responsiveness to microenvironment stimuli. In the present study, the effects of lipopolysaccharide (LPS) and lipoteichoic acid (LTA), as well-known mediators of inflammation, on cancerous behavior of three prostate tumor cells, LNCaP, PC3 and DU145, were investigated. METHODS: Expression of TLR1-10, CD14 and MyD88 transcripts was investigated by RT-PCR. Protein expression of TLR2 and 4 was scrutinized by flow cytometry, immunofluorescent staining and Western blotting. Experiments were set up to assess the effects of LPS and LTA at different concentrations and times on cell proliferation, extracellular matrix invasion, adhesion and cytokine production. RESULTS: We showed that prostate cancer cell lines differentially express TLR1-10, MyD88 and CD14 transcripts. DU145 failed to express TLR4 gene. Positively-identified TLR2 protein in all prostate cancer cells and TLR4 protein in PC3 and LNCaP by Western blotting was not accompanied by cell surface expression, as judged by flow cytometry. Immunofluorescent staining clearly demonstrated predominantly perinuclear localization of TLR2 and TLR4. LTA activation of all prostate cancer cells significantly increased cell proliferation. Regardless of lacking TLR4, DU145 cells proliferated in response to LPS treatment. While LPS caused increased invasiveness of LNCaP, invasive capacity of PC3 was significantly reduced after LPS or LTA stimulation. Stimulation of all prostate tumor cells with LTA was associated with increased cell adhesion and IL-8 production. IL-6 production, however, was differentially regulated by LPS stimulation in prostate tumor cells. CONCLUSION: The data shows that cancer cells originated from the same histologically origin exhibit heterogeneous response to the same TLR ligand. Therefore, a thorough and comprehensive judgment on how and to what extent a particular cancer is affected by TLR agonist could not be inferred by studying an individual cell line.

11.
J Assist Reprod Genet ; 31(1): 121-4, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24189965

ABSTRACT

PURPOSE: To compare plasminogen activator inhibitor type1 (PAI-1) mutation rates in different groups of patients with the record of recurrent miscarriage (RM) or implantation failure (IF) with special emphasis on the number of missed pregnancies and/or implantation failures (RM ≥ 2, IF ≥ 2, RM + IF ≥ 2, RM ≥ 3, IF ≥ 3 and RM + IF ≥ 3). METHOD: Case-control study from PCR products and RFLP data of DNA from blood of patients who referred to the infertility clinic including 595 patients (421 RM ≥ 2, 119 IF ≥ 2 and 55 RM + IF ≥ 2) as the case groups and 100 healthy women as the control group. RESULTS: All six different subgroups of patients showed increased frequencies of the mutant allele (4G) in comparison to the control group (p < 0.001) suggesting a role for PAI-1 mutation in RM and IF. CONCLUSIONS: The different patient subgroups suffer similar rates of risk in developing RM and IF when compared to controls.


Subject(s)
Abortion, Habitual/genetics , Embryo Implantation/genetics , Embryo Loss/genetics , Mutation , Plasminogen Activator Inhibitor 1/genetics , Abortion, Habitual/epidemiology , Adult , Case-Control Studies , Embryo Loss/epidemiology , Female , Genetic Association Studies , Genetic Predisposition to Disease , Humans , Polymorphism, Restriction Fragment Length , Pregnancy , Young Adult
12.
Iran J Public Health ; 43(11): 1569-75, 2014 Nov.
Article in English | MEDLINE | ID: mdl-26060726

ABSTRACT

BACKGROUND: Varicella zoster virus (VZV) is a member of herpes family viruses, which causes varicella (chickenpox) after primary infection and herpes zoster (shingles) because of latent virus reactivation from dorsal root ganglia. Generally, prevalence of varicella antibodies increases with age. We aimed to compare the prevalence of anti-VZV antibody in children under seven years old, in order to obtain a preliminarily picture of general presence of these antibodies to design an immunization plan. METHODS: In this cross-sectional study, performed from September 2011 to September 2012 in Tehran, Iran, 267 serum samples including sera from 7 month old infants, n= 87; 18 month old children, n= 86; and 6 year old children, n= 94 were assessed for the presence of specific IgG antibodies against VZV, using ELISA technique. RESULTS: 4.6% of 7 month, 12.8% of 18 month and 21.3% of 6-year-old children were seropositive. No relation was found between demographic variables (e.g. age and birth weight) and seropositivity in these age groups. VZV antibodies increased with age. Serum levels of varicella antibodies were elevated in 18 months old compared to 7 months old children, significantly (P < 0.001). CONCLUSION: In view of the significant elevation of VZV antibodies in children from 7 months to 18 months of age and rate of seronegative children, our results support the necessity of varicella immunization between 7 and 18 months of age in order to prevent viral infection.

13.
Iran Red Crescent Med J ; 15(4): 340-4, 2013 Apr.
Article in English | MEDLINE | ID: mdl-24083010

ABSTRACT

BACKGROUND: Chlamydia trachomatis and Mycoplasma genitalium infections are the most prevalent sexually transmitted bacterial infections in the world that cause urogenital infections in both men and women. It appears that infertility is a complication of these infections. OBJECTIVE: This study was designed to estimate the prevalence of Chlamydia trachomatis and Mycoplasma genitalium in symptomatic and asymptomatic men and to assess risk factors associated with infection. PATIENTS AND METHODS: Urine specimens were collected from 200 men; 100 of them were symptomatic and 100 asymptomatic. Samples were examined by PCR to detect the infections. RESULTS: C. trachomatis was detected in 20% of symptomatic and in 4% of asymptomatic men (P < 0.001). The prevalence of M. genitalium was revealed to be 12% and 2% in symptomatic and asymptomatic men, respectively (P < 0.01). Four of 100 men in the symptomatic group were infected with both organisms. C. trachomatis infection was associated with dysuria, urethral discharge, testicular swelling, and genital ulcer (P < 0.05). M. genitalium infection was related with dysuria, testis inflammation, pelvic pain and low educational level (P < 0.05). Furthermore, the prevalence of infections at ages 30-39 years was more than other ages. CONCLUSIONS: Considering the role of these bacteria in urogenital infections, a screening test is recommended. Since the PCR assay is a highly sensitive and specific assay for the detection of these bacteria in male urine specimens, it provides a noninvasive technique for routine screening.

14.
Hum Vaccin Immunother ; 9(6): 1316-22, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23442608

ABSTRACT

The present study was undertaken to compare the immunogenicity and reactogenicity of two diphtheria-tetanus-whole cell pertussis (DTwP) vaccines administered to Iranian preschool children. In this randomized, double-blind and multicenter prospective study, 672 children aged 4-6 y were administered with either a local DTwP vaccine (DTwP-Local) (n = 337) or a commercial vaccine (DTwP-Pasteur) (n = 335). All subjects received DTwP vaccine at 4-6 y of age, following the national immunization schedule of Iran. Blood samples were collected before and 2-4 weeks after the vaccination. Immunogenicity of each vaccine was assessed by ELISA using commercial kits. Reactogenicity was assessed by the parents for seven days post-booster using diary cards. The geometric mean titers (GMTs) of the antibodies induced against diphtheria and tetanus by DTwP-Local were 7.7 and 9.4 IU/ml and those of DTwP-Pasteur were 8.2 and 8.6 IU/ml, respectively. There was no significant difference between the immunogenicity of the two vaccines against diphtheria and tetanus. The GMTs of antibodies produced against pertussis were 30.2 EU/ml for DTwP-Local and 47.9 EU/ml for DTwP-Pasteur vaccines (p<0.001). Pain and fever (axillary temperature>37.5°C) were the most frequent local and systemic reactions observed after the vaccination. All local and systemic reactions observed after vaccination were significantly higher in subjects immunized with DTwP-Local vaccine. Immunogenicity against diphtheria and tetanus was similar for the two vaccines, but immunogenicity of the local vaccine against pertussis was significantly less efficient than that of DTwP-Pasteur. This difference and the higher side effects of the DTwP-Local vaccine could be due to the bacterial strain or the preparation or formulation protocol of the local pertussis vaccine.


Subject(s)
Diphtheria-Tetanus-Pertussis Vaccine/adverse effects , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Antibodies, Bacterial , Child , Child, Preschool , Diphtheria-Tetanus-Pertussis Vaccine/administration & dosage , Double-Blind Method , Drug-Related Side Effects and Adverse Reactions/epidemiology , Drug-Related Side Effects and Adverse Reactions/pathology , Enzyme-Linked Immunosorbent Assay , Female , Fever/chemically induced , Fever/epidemiology , Humans , Iran , Male , Pain/chemically induced , Pain/epidemiology , Prospective Studies
15.
Hybridoma (Larchmt) ; 31(5): 372-7, 2012 Oct.
Article in English | MEDLINE | ID: mdl-23098305

ABSTRACT

Leptin is an important protein that regulates energy storage and homeostasis in humans and animals. Leptin deficiency results in various abnormalities such as diabetes, obesity, and infertility. Producing a high affinity monoclonal antibody against human leptin provides an important tool to monitor and trace leptin function in different biological fluids. In this study, recombinant human leptin was conjugated to KLH and injected into mice. After immunization, mouse myeloma SP2/0 cells were fused with murine splenocytes followed by selection of antibody-producing hybridoma cells. After screening of different hybridoma colonies by ELISA, a high affinity antibody was selected and purified by affinity chromatography. The affinity constant of the antibody was measured by ELISA. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibody. The anti-leptin antibody had a high affinity (around 1.13 × 10(-9) M) for its antigen. The saturation of the antibody with leptin (20 moles leptin per 1 mole antibody) in Western blot analysis proved that the antibody had specific binding to its antigen. Immunocytochemistry and flow cytometry on JEG-3 (human placental choriocarcinoma cell) cells revealed that the anti-leptin antibody recognized intracellular leptin. In conclusion, we report here the production and characterization of a murine anti-leptin antibody with high affinity for human leptin.


Subject(s)
Antibodies, Monoclonal/immunology , Antibody Specificity , Hemocyanins/immunology , Immunoconjugates/immunology , Leptin/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Antibody Affinity , Blotting, Western , Cell Line, Tumor , Chromatography, Affinity , Enzyme-Linked Immunosorbent Assay , Female , Hemocyanins/chemistry , Humans , Hybridomas/immunology , Immunization , Immunoconjugates/administration & dosage , Immunoconjugates/chemistry , Kinetics , Leptin/administration & dosage , Mice , Mice, Inbred BALB C , Pregnancy , Recombinant Proteins/administration & dosage , Recombinant Proteins/immunology
16.
J Reprod Infertil ; 13(2): 89-94, 2012 Apr.
Article in English | MEDLINE | ID: mdl-23926530

ABSTRACT

BACKGROUND: Recurrent pregnancy loss is (RPL) a heterogeneous condition. While the role of acquired thrombophilia has been accepted as an etiology for RPL, the contribution of specific inherited thrombophilic gene polymorphisms to the disorder has been remained controversial. METHODS: One hundred women with a history of two or more consecutive abortions and 100 women with at least two live births and no miscarriages were included in the study and evaluated for the presence of 11 thrombophilic gene polymorphisms (Factor V LEIDEN, Factor V 4070 A/G, Factor V 5279 A/G, Factor XIII 103 G/T, Factor XIII 614 A/T, Factor XIII 1694 C/T, PAI-1 -675 4G/5G, ITGB3 1565 T/C, ß-Fibrinogen -455G/A, MTHFR 677 C/T, MTHFR 1298 A/C) using PCR-RFLP technique. The data were statistically analyzed using Mann-Whitney test and logistic regression model. RESULTS: There was no relation between factor XIII 103G/T gene polymorphism with increased risk of RPL. However, the other 10 gene polymorphisms were found to be associated with increased/decreased risk of RPL. Multiple logistic regression model for analyzing the simultaneous effects of these polymorphisms on the risk of RPL showed that six of these 11 polymorphisms (Factor V 1691G/A, Factor V 5279A/G, Factor XIII 614A/T, ß-Fibrinogen -455G/A, ITGB3 1565T/C, and MTHFR 1298A/ C) were associated with RPL. CONCLUSION: It is possible to calculate the risk of abortion in a patient with RPL by determining only six of the 10 polymorphisms that are individually associated with RPL.

17.
J Nanobiotechnology ; 9: 31, 2011 Aug 08.
Article in English | MEDLINE | ID: mdl-21824404

ABSTRACT

BACKGROUND: Several materials are available in the market that work on the principle of protein magnetic fishing by their histidine (His) tags. Little information is available on their performance and it is often quoted that greatly improved purification of histidine-tagged proteins from crude extracts could be achieved. While some commercial magnetic matrices could be used successfully for purification of several His-tagged proteins, there are some which have been proved to operate just for a few extent of His-tagged proteins. Here, we address quantitative evaluation of three commercially available Nickel nanomagnetic beads for purification of two His-tagged proteins expressed in Escherichia coli and present helpful hints for optimized purification of such proteins and preparation of nanomagnetisable matrices. RESULTS: Marked differences in the performance of nanomagnetic matrices, principally on the basis of their specific binding capacity, recovery profile, the amount of imidazole needed for protein elution and the extent of target protein loss and purity were obtained. Based on the aforesaid criteria, one of these materials featured the best purification results (SiMAG/N-NTA/Nickel) for both proteins at the concentration of 4 mg/ml, while the other two (SiMAC-Nickel and SiMAG/CS-NTA/Nickel) did not work well with respect to specific binding capacity and recovery profile. CONCLUSIONS: Taken together, functionality of different types of nanomagnetic matrices vary considerably. This variability may not only be dependent upon the structure and surface chemistry of the matrix which in turn determine the affinity of interaction, but, is also influenced to a lesser extent by the physical properties of the protein itself. Although the results of the present study may not be fully applied for all nanomagnetic matrices, but provide a framework which could be used to profiling and quantitative evaluation of other magnetisable matrices and also provide helpful hints for those researchers facing same challenge.


Subject(s)
Magnetite Nanoparticles/chemistry , Nickel/chemistry , Recombinant Proteins/isolation & purification , Histidine/chemistry
18.
Am J Reprod Immunol ; 66(2): 149-56, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21241403

ABSTRACT

PROBLEM: To identify the associations of the plasminogen activator inhibitor-1 (PAI-1) -675 4G/5G, beta fibrinogen (BF) -455G/A, integrin beta 3 (ITGB3) 1565T/C, and methylenetetrahydrofolate reductase (MTHFR) 677C/T and 1298A/C polymorphisms with recurrent pregnancy loss (RPL). METHOD OF STUDY: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were performed to assess the frequency of five candidate genetic risk factors for RPL, and the frequencies of the polymorphisms were calculated and compared between case and control groups. RESULTS: The BF -455G/A, MTHFR 677C/T, and 1298A/C polymorphisms were found to be positively, and ITGB3 1565T/C polymorphism negatively, associated with RPL. Homozygosity but not heterozygosity for PAI-1 -675 4G/5G polymorphism was significantly higher in patients with RPL than in the control group. The presence of both mutations of MTHFR genes highly increased the risk of RPL. CONCLUSION: The data highlight the importance of thrombophilia screening in patients with RPL.


Subject(s)
Abortion, Habitual/genetics , Fibrinogen/genetics , Integrin beta3/genetics , Methylenetetrahydrofolate Reductase (NADPH2)/genetics , Plasminogen Activator Inhibitor 1/genetics , Polymorphism, Genetic/genetics , Female , Humans , Polymerase Chain Reaction , Pregnancy , Risk Factors
19.
Am J Reprod Immunol ; 64(3): 212-7, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20384622

ABSTRACT

PROBLEM: among important suspected causes of thrombophilia in women with recurrent pregnancy loss (RPL) are the polymorphisms of coagulation factor XIII (FXIII) gene. We performed a case-control study on the association between three polymorphisms of factor XIII (FXIII G103T, FXIII A614T and FXIII C1694T) and RPL in Iranian women. METHOD OF STUDY: DNA samples from peripheral blood of 100 female patients with at least two recurrent abortions, as case group, and 100 healthy women with history of at least two successful deliveries were subjected to PCR-RFLP, and the frequencies of the polymorphisms were calculated and compared between the two groups. RESULTS: the prevalence of FXIII G103T polymorphism was 29% in the case group and 17% in the control group (P = 0.158). The frequencies of FXIII A614T and FXIII C1694T were 84% and 66% in the case group and 48% and 31% in the control group (P <0.001 and P < 0.001), respectively. The two latter polymorphisms are associated with RPL in Iranian women and increase the risk of RPL. A correlation was also found between FXIII A614T and FXIII C1694T polymorphisms (P < 0.001). CONCLUSION: we suggest the evaluation of FXIII A614T and FXIII C1694T polymorphisms in women with RPL.


Subject(s)
Abortion, Habitual/genetics , Factor XIII/genetics , Abortion, Habitual/epidemiology , Abortion, Habitual/physiopathology , Case-Control Studies , DNA Mutational Analysis , Female , Gene Frequency , Genetic Association Studies , Genetic Predisposition to Disease , Humans , Iran , Polymorphism, Genetic , Pregnancy , Risk Factors , Thrombophilia
20.
Iran J Allergy Asthma Immunol ; 8(2): 85-93, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19671937

ABSTRACT

Universal vaccination of neonates and children against diphtheria, tetanus and pertussis has had a tremendous impact on the control of these infectious diseases worldwide. Immunization by the triple diphtheria, tetanus and whole cell pertussis vaccine (DTwP) has been applied in Iran for almost 50 years. Periodic assessment of immunogenicity of this vaccine is an important aspect of successful mass vaccination programs. The present study was performed to assess the antibody response against tetanus, diphtheria and pertussis in a group of Iranian infants vaccinated with a local DTwP vaccine. In this prospective study, 330 infants received primary vaccination at 2, 4 and 6 months of age with DTwP vaccine manufactured by Razi Institute of Iran. Blood samples were taken 2-4 weeks after the third dose to assess seroprotection and geometric mean titers ( GMT) of specific antibodies. Among the 283 infants who completed the vaccination course, 98.2% and 100% developed antibodies against diphtheria and tetanus, respectively. The GMT of antibodies to tetanus, diphtheria and pertussis, were 2.09 IU/ml, 2.08 IU/ml and 8.73 EU/ml, respectively. Comparison of the results obtained from this study with those from previous studies performed in other countries revealed a similar GMT and protection rates for diphtheria and tetanus components. In the absence of well-established serological criteria, judgment about protection rate against pertussis has not been possible. A prospective vaccination study using the local DTwP vaccine in parallel to a WHO approved standard vaccine, could enable assessment of immunogenicity of the pertussis component.


Subject(s)
Antibodies, Bacterial/blood , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Diphtheria/prevention & control , Tetanus/prevention & control , Whooping Cough/prevention & control , Antibody Formation , Antigens, Bacterial/immunology , Diphtheria/blood , Diphtheria/immunology , Diphtheria/microbiology , Diphtheria-Tetanus-Pertussis Vaccine/blood , Female , Humans , Infant , Iran , Linear Models , Male , Prospective Studies , Tetanus/blood , Tetanus/immunology , Tetanus/microbiology , Whooping Cough/blood , Whooping Cough/immunology , Whooping Cough/microbiology
SELECTION OF CITATIONS
SEARCH DETAIL
...