1.Detection of Eosinophilic Cell-free Granules Based on Expression of CCR3 and MBP Markers in Esophageal Biopsy Specimens of Patients with Suspected and Confirmed Eosinophilic Esophagitis
Saba EBRAHIMI ; Saghi KHATAMI ; Fatemeh Elham MAHJOUB ; Maryam MONAJEMZADEH ; Azizollah YOUSEFI ; Amirhossein HOSSEINI ; Majid KHOSHMIRSAFA ; Fatemeh.S. MOUSAVINASAB ; Mahboubeh MANSOURI ; Mehdi SHABANI ; Mehrnaz MESDAGHI
Pediatric Gastroenterology, Hepatology & Nutrition 2025;28(1):19-26
Purpose:
Eosinophilic esophagitis (EoE) is the most well-known eosinophilic gastrointestinal disorder (EGID) characterized by the presence of a high number eosinophils within the esophageal epithelium and the clinical signs. Biopsies of patients with suspected EoE may not show a high number of eosinophils, however the presence of granules may help with the diagnosis. This study aims to evaluate the presence of cell-free eosinophil granules in the esophageal tissue of patients with suspected and confirmed EoE to accelerate the diagnosis and treatment of patients with low eosinophil count.
Methods:
Fifteen patients with confirmed EoE and 15 suspected of EoE were included in this study. Patients' esophageal tissue biopsies were stained using immunohistochemistry (IHC) to identify eosinophils and their cell-free granules. For testing, anti-major basic protein (MBP) and anti-chemokine receptor type 3 (CCR3) were used as primary antibodies and a double-staining kit containing secondary antibodies conjugated to the enzyme and related chromogens were used.
Results:
Cell-free granules with different degrees were observed in 53.3% and 93.3% of suspected and confirmed EoE samples, respectively. Furthermore, in esophageal biopsy of 73.3% of patients with suspected and 93.3% of patients with a definitive diagnosis of EoE, basal layer hyperplasia (BLH) was recognized.
Conclusion
The results of the present study showed that IHC can be applied to detect cell-free eosinophil granules in esophageal tissue. Observation of granules and basal cell hyperplasia in biopsies of suspected EoE patients whose eosinophil count is below the threshold can be valuable findings to make a definitive diagnosis for these patients.
2.Detection of Eosinophilic Cell-free Granules Based on Expression of CCR3 and MBP Markers in Esophageal Biopsy Specimens of Patients with Suspected and Confirmed Eosinophilic Esophagitis
Saba EBRAHIMI ; Saghi KHATAMI ; Fatemeh Elham MAHJOUB ; Maryam MONAJEMZADEH ; Azizollah YOUSEFI ; Amirhossein HOSSEINI ; Majid KHOSHMIRSAFA ; Fatemeh.S. MOUSAVINASAB ; Mahboubeh MANSOURI ; Mehdi SHABANI ; Mehrnaz MESDAGHI
Pediatric Gastroenterology, Hepatology & Nutrition 2025;28(1):19-26
Purpose:
Eosinophilic esophagitis (EoE) is the most well-known eosinophilic gastrointestinal disorder (EGID) characterized by the presence of a high number eosinophils within the esophageal epithelium and the clinical signs. Biopsies of patients with suspected EoE may not show a high number of eosinophils, however the presence of granules may help with the diagnosis. This study aims to evaluate the presence of cell-free eosinophil granules in the esophageal tissue of patients with suspected and confirmed EoE to accelerate the diagnosis and treatment of patients with low eosinophil count.
Methods:
Fifteen patients with confirmed EoE and 15 suspected of EoE were included in this study. Patients' esophageal tissue biopsies were stained using immunohistochemistry (IHC) to identify eosinophils and their cell-free granules. For testing, anti-major basic protein (MBP) and anti-chemokine receptor type 3 (CCR3) were used as primary antibodies and a double-staining kit containing secondary antibodies conjugated to the enzyme and related chromogens were used.
Results:
Cell-free granules with different degrees were observed in 53.3% and 93.3% of suspected and confirmed EoE samples, respectively. Furthermore, in esophageal biopsy of 73.3% of patients with suspected and 93.3% of patients with a definitive diagnosis of EoE, basal layer hyperplasia (BLH) was recognized.
Conclusion
The results of the present study showed that IHC can be applied to detect cell-free eosinophil granules in esophageal tissue. Observation of granules and basal cell hyperplasia in biopsies of suspected EoE patients whose eosinophil count is below the threshold can be valuable findings to make a definitive diagnosis for these patients.
3.Oxidized Mannan: A Novel Adjuvant Candidate for Enhancing Immune Responses in Veterinary Rabies Vaccine
Rajab MARDANI ; Anahita BAHMANJE ; Yousef Cheraghi KAZERONI ; Fereydoon KHOSHROO ; Bahram ROSHANAIE ; Tahereh SADEGHCHE ; Kourosh PAJAIE ; Seyed Nezamedin HOSSEINI ; Delaram DOROUD ; Maryam SHAHALI
Chonnam Medical Journal 2025;61(2):84-89
Rabies continues to pose a serious public health threat worldwide, with vaccination being the most effective means of prevention. However, commercially available inactivated rabies vaccines often require multiple doses and lack potent adjuvants to enhance their efficacy. This study aimed to investigate the coupling of whole inactivated rabies virus to mannan under oxidizing conditions to improve immune responses against a standard rabies vaccine. We explored the conjugation of whole inactivated rabies virus with oxidized mannan (Rab-OxMan) to enhance immune responses. Mice were immunized intraperitoneally with 350 µg of the Rab-OxMan formulation on days 1 and 7. Two weeks after immunization, serum samples were collected to measure levels of IgG, IgM, and TNF- using ELISA. The vaccine’s potency was also evaluated using the National Institutes of Health (NIH) assay. Our findings showed a significant increase in IgG levels and a decrease in IgM levels in the Rab-OxMan group compared to the Alum-adjuvanted vaccine group (p<0.05). Additionally, TNF- levels were notably higher in the Rab-OxMan group (p<0.05). Statistical analysis revealed that IgG levels had the highest sensitivity and specificity, with a significant correlation between the measured variables. Importantly, the Rab-OxMan formulation provided 1.8 times greater protection in challenge tests compared to the alum-adjuvanted group. This study is the first to demonstrate that oxidized mannan can serve as a novel adjuvant for veterinary rabies vaccines. The results highlight significant improvements in the immunogenicity and efficacy of rabies vaccines, suggesting a promising strategy for enhancing rabies prevention and potentially reducing the incidence of this deadly disease.
4.Detection of Eosinophilic Cell-free Granules Based on Expression of CCR3 and MBP Markers in Esophageal Biopsy Specimens of Patients with Suspected and Confirmed Eosinophilic Esophagitis
Saba EBRAHIMI ; Saghi KHATAMI ; Fatemeh Elham MAHJOUB ; Maryam MONAJEMZADEH ; Azizollah YOUSEFI ; Amirhossein HOSSEINI ; Majid KHOSHMIRSAFA ; Fatemeh.S. MOUSAVINASAB ; Mahboubeh MANSOURI ; Mehdi SHABANI ; Mehrnaz MESDAGHI
Pediatric Gastroenterology, Hepatology & Nutrition 2025;28(1):19-26
Purpose:
Eosinophilic esophagitis (EoE) is the most well-known eosinophilic gastrointestinal disorder (EGID) characterized by the presence of a high number eosinophils within the esophageal epithelium and the clinical signs. Biopsies of patients with suspected EoE may not show a high number of eosinophils, however the presence of granules may help with the diagnosis. This study aims to evaluate the presence of cell-free eosinophil granules in the esophageal tissue of patients with suspected and confirmed EoE to accelerate the diagnosis and treatment of patients with low eosinophil count.
Methods:
Fifteen patients with confirmed EoE and 15 suspected of EoE were included in this study. Patients' esophageal tissue biopsies were stained using immunohistochemistry (IHC) to identify eosinophils and their cell-free granules. For testing, anti-major basic protein (MBP) and anti-chemokine receptor type 3 (CCR3) were used as primary antibodies and a double-staining kit containing secondary antibodies conjugated to the enzyme and related chromogens were used.
Results:
Cell-free granules with different degrees were observed in 53.3% and 93.3% of suspected and confirmed EoE samples, respectively. Furthermore, in esophageal biopsy of 73.3% of patients with suspected and 93.3% of patients with a definitive diagnosis of EoE, basal layer hyperplasia (BLH) was recognized.
Conclusion
The results of the present study showed that IHC can be applied to detect cell-free eosinophil granules in esophageal tissue. Observation of granules and basal cell hyperplasia in biopsies of suspected EoE patients whose eosinophil count is below the threshold can be valuable findings to make a definitive diagnosis for these patients.
5.Oxidized Mannan: A Novel Adjuvant Candidate for Enhancing Immune Responses in Veterinary Rabies Vaccine
Rajab MARDANI ; Anahita BAHMANJE ; Yousef Cheraghi KAZERONI ; Fereydoon KHOSHROO ; Bahram ROSHANAIE ; Tahereh SADEGHCHE ; Kourosh PAJAIE ; Seyed Nezamedin HOSSEINI ; Delaram DOROUD ; Maryam SHAHALI
Chonnam Medical Journal 2025;61(2):84-89
Rabies continues to pose a serious public health threat worldwide, with vaccination being the most effective means of prevention. However, commercially available inactivated rabies vaccines often require multiple doses and lack potent adjuvants to enhance their efficacy. This study aimed to investigate the coupling of whole inactivated rabies virus to mannan under oxidizing conditions to improve immune responses against a standard rabies vaccine. We explored the conjugation of whole inactivated rabies virus with oxidized mannan (Rab-OxMan) to enhance immune responses. Mice were immunized intraperitoneally with 350 µg of the Rab-OxMan formulation on days 1 and 7. Two weeks after immunization, serum samples were collected to measure levels of IgG, IgM, and TNF- using ELISA. The vaccine’s potency was also evaluated using the National Institutes of Health (NIH) assay. Our findings showed a significant increase in IgG levels and a decrease in IgM levels in the Rab-OxMan group compared to the Alum-adjuvanted vaccine group (p<0.05). Additionally, TNF- levels were notably higher in the Rab-OxMan group (p<0.05). Statistical analysis revealed that IgG levels had the highest sensitivity and specificity, with a significant correlation between the measured variables. Importantly, the Rab-OxMan formulation provided 1.8 times greater protection in challenge tests compared to the alum-adjuvanted group. This study is the first to demonstrate that oxidized mannan can serve as a novel adjuvant for veterinary rabies vaccines. The results highlight significant improvements in the immunogenicity and efficacy of rabies vaccines, suggesting a promising strategy for enhancing rabies prevention and potentially reducing the incidence of this deadly disease.
6.Detection of Eosinophilic Cell-free Granules Based on Expression of CCR3 and MBP Markers in Esophageal Biopsy Specimens of Patients with Suspected and Confirmed Eosinophilic Esophagitis
Saba EBRAHIMI ; Saghi KHATAMI ; Fatemeh Elham MAHJOUB ; Maryam MONAJEMZADEH ; Azizollah YOUSEFI ; Amirhossein HOSSEINI ; Majid KHOSHMIRSAFA ; Fatemeh.S. MOUSAVINASAB ; Mahboubeh MANSOURI ; Mehdi SHABANI ; Mehrnaz MESDAGHI
Pediatric Gastroenterology, Hepatology & Nutrition 2025;28(1):19-26
Purpose:
Eosinophilic esophagitis (EoE) is the most well-known eosinophilic gastrointestinal disorder (EGID) characterized by the presence of a high number eosinophils within the esophageal epithelium and the clinical signs. Biopsies of patients with suspected EoE may not show a high number of eosinophils, however the presence of granules may help with the diagnosis. This study aims to evaluate the presence of cell-free eosinophil granules in the esophageal tissue of patients with suspected and confirmed EoE to accelerate the diagnosis and treatment of patients with low eosinophil count.
Methods:
Fifteen patients with confirmed EoE and 15 suspected of EoE were included in this study. Patients' esophageal tissue biopsies were stained using immunohistochemistry (IHC) to identify eosinophils and their cell-free granules. For testing, anti-major basic protein (MBP) and anti-chemokine receptor type 3 (CCR3) were used as primary antibodies and a double-staining kit containing secondary antibodies conjugated to the enzyme and related chromogens were used.
Results:
Cell-free granules with different degrees were observed in 53.3% and 93.3% of suspected and confirmed EoE samples, respectively. Furthermore, in esophageal biopsy of 73.3% of patients with suspected and 93.3% of patients with a definitive diagnosis of EoE, basal layer hyperplasia (BLH) was recognized.
Conclusion
The results of the present study showed that IHC can be applied to detect cell-free eosinophil granules in esophageal tissue. Observation of granules and basal cell hyperplasia in biopsies of suspected EoE patients whose eosinophil count is below the threshold can be valuable findings to make a definitive diagnosis for these patients.
7.Oxidized Mannan: A Novel Adjuvant Candidate for Enhancing Immune Responses in Veterinary Rabies Vaccine
Rajab MARDANI ; Anahita BAHMANJE ; Yousef Cheraghi KAZERONI ; Fereydoon KHOSHROO ; Bahram ROSHANAIE ; Tahereh SADEGHCHE ; Kourosh PAJAIE ; Seyed Nezamedin HOSSEINI ; Delaram DOROUD ; Maryam SHAHALI
Chonnam Medical Journal 2025;61(2):84-89
Rabies continues to pose a serious public health threat worldwide, with vaccination being the most effective means of prevention. However, commercially available inactivated rabies vaccines often require multiple doses and lack potent adjuvants to enhance their efficacy. This study aimed to investigate the coupling of whole inactivated rabies virus to mannan under oxidizing conditions to improve immune responses against a standard rabies vaccine. We explored the conjugation of whole inactivated rabies virus with oxidized mannan (Rab-OxMan) to enhance immune responses. Mice were immunized intraperitoneally with 350 µg of the Rab-OxMan formulation on days 1 and 7. Two weeks after immunization, serum samples were collected to measure levels of IgG, IgM, and TNF- using ELISA. The vaccine’s potency was also evaluated using the National Institutes of Health (NIH) assay. Our findings showed a significant increase in IgG levels and a decrease in IgM levels in the Rab-OxMan group compared to the Alum-adjuvanted vaccine group (p<0.05). Additionally, TNF- levels were notably higher in the Rab-OxMan group (p<0.05). Statistical analysis revealed that IgG levels had the highest sensitivity and specificity, with a significant correlation between the measured variables. Importantly, the Rab-OxMan formulation provided 1.8 times greater protection in challenge tests compared to the alum-adjuvanted group. This study is the first to demonstrate that oxidized mannan can serve as a novel adjuvant for veterinary rabies vaccines. The results highlight significant improvements in the immunogenicity and efficacy of rabies vaccines, suggesting a promising strategy for enhancing rabies prevention and potentially reducing the incidence of this deadly disease.
8.Detection of Eosinophilic Cell-free Granules Based on Expression of CCR3 and MBP Markers in Esophageal Biopsy Specimens of Patients with Suspected and Confirmed Eosinophilic Esophagitis
Saba EBRAHIMI ; Saghi KHATAMI ; Fatemeh Elham MAHJOUB ; Maryam MONAJEMZADEH ; Azizollah YOUSEFI ; Amirhossein HOSSEINI ; Majid KHOSHMIRSAFA ; Fatemeh.S. MOUSAVINASAB ; Mahboubeh MANSOURI ; Mehdi SHABANI ; Mehrnaz MESDAGHI
Pediatric Gastroenterology, Hepatology & Nutrition 2025;28(1):19-26
Purpose:
Eosinophilic esophagitis (EoE) is the most well-known eosinophilic gastrointestinal disorder (EGID) characterized by the presence of a high number eosinophils within the esophageal epithelium and the clinical signs. Biopsies of patients with suspected EoE may not show a high number of eosinophils, however the presence of granules may help with the diagnosis. This study aims to evaluate the presence of cell-free eosinophil granules in the esophageal tissue of patients with suspected and confirmed EoE to accelerate the diagnosis and treatment of patients with low eosinophil count.
Methods:
Fifteen patients with confirmed EoE and 15 suspected of EoE were included in this study. Patients' esophageal tissue biopsies were stained using immunohistochemistry (IHC) to identify eosinophils and their cell-free granules. For testing, anti-major basic protein (MBP) and anti-chemokine receptor type 3 (CCR3) were used as primary antibodies and a double-staining kit containing secondary antibodies conjugated to the enzyme and related chromogens were used.
Results:
Cell-free granules with different degrees were observed in 53.3% and 93.3% of suspected and confirmed EoE samples, respectively. Furthermore, in esophageal biopsy of 73.3% of patients with suspected and 93.3% of patients with a definitive diagnosis of EoE, basal layer hyperplasia (BLH) was recognized.
Conclusion
The results of the present study showed that IHC can be applied to detect cell-free eosinophil granules in esophageal tissue. Observation of granules and basal cell hyperplasia in biopsies of suspected EoE patients whose eosinophil count is below the threshold can be valuable findings to make a definitive diagnosis for these patients.
9.Preventive efficacy of hydrocortisone enema for radiation proctitis in rectal cancer patients undergoing short-course radiotherapy: a phase II randomized placebo-controlled clinical trial
Mohammad MOHAMMADIANPANAH ; Maryam TAZANG ; Nam Phong NGUYEN ; Niloofar AHMADLOO ; Shapour OMIDVARI ; Ahmad MOSALAEI ; Mansour ANSARI ; Hamid NASROLLAHI ; Behnam KADKHODAEI ; Nezhat KHANJANI ; Seyed Vahid HOSSEINI
Annals of Coloproctology 2024;40(5):506-514
Purpose:
This study aimed to investigate the efficacy of hydrocortisone enema in preventing radiation proctitis in patients with rectal cancer undergoing short-course radiotherapy (SCRT).
Methods:
This phase II randomized controlled trial enrolled patients with newly diagnosed locally advanced rectal cancer (clinically staged T3–4 and/or N1–2M0). Participants received a median of 4 cycles of neoadjuvant chemotherapy (capecitabine plus oxaliplatin) followed by 3-dimensional conformal SCRT (25 Gy in 5 fractions). Patients were randomly assigned to receive either a hydrocortisone enema (n=50) or a placebo (n=51) once daily for 5 consecutive days during SCRT. The primary endpoint was the incidence and severity of acute proctitis.
Results:
Of the 111 eligible patients, 101 were included in the study. Baseline characteristics, including sex, age, performance status, and tumor location, were comparable across the treatment arms. None of the patients experienced grade 4 acute gastrointestinal toxicity or had to discontinue treatment due to treatment-related adverse effects. Patients in the hydrocortisone arm experienced significantly less severe proctitis (P<0.001), diarrhea (P=0.023), and rectal pain (P<0.001) than those in the placebo arm. Additionally, the duration of acute gastrointestinal toxicity following SCRT was significantly shorter in patients receiving hydrocortisone (P<0.001).
Conclusion
Hydrocortisone enema was associated with a significant reduction in the severity of proctitis, diarrhea, and rectal pain compared to placebo. Additionally, patients treated with hydrocortisone experienced shorter durations of gastrointestinal toxicity following SCRT. This study highlights the potential benefits of hydrocortisone enema in managing radiation-induced toxicity in rectal cancer patients undergoing radiotherapy.
10.In vivo and in vitro sperm production: an overview of the challenges and advances in male fertility restoration
Zahra BASHIRI ; Seyed Jamal HOSSEINI ; Maryam SALEM ; Morteza KORUJI
Clinical and Experimental Reproductive Medicine 2024;51(3):171-180
Male infertility can be caused by genetic anomalies, endocrine disorders, inflammation, and exposure to toxic chemicals or gonadotoxic treatments. Therefore, several recent studies have concentrated on the preservation and restoration of fertility to enhance the quality of life for affected individuals. It is currently recommended to biobank the tissue extracted from testicular biopsies to provide a later source of spermatogonial stem cells (SSCs). Another successful approach has been the in vitro production of haploid male germ cells. The capacity of SSCs to transform into sperm, as in testicular tissue transplantation, SSC therapy, and in vitro or ex vivo spermatogenesis, makes them ideal candidates for in vivo fertility restoration. The transplantation of SSCs or testicular tissue to regenerate spermatogenesis and create embryos has been achieved in nonhuman mammal species. Although the outcomes of human trials have yet to be released, this method may soon be approved for clinical use in humans. Furthermore, regenerative medicine techniques that develop tissue or cells on organic or synthetic scaffolds enriched with bioactive molecules have also gained traction. All of these methods are now in different stages of experimentation and clinical trials. However, thanks to rigorous studies on the safety and effectiveness of SSC-based reproductive treatments, some of these techniques may be clinically available in upcoming decades.

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