Neven A. Ebrahim;Hussein Abdelaziz Abdalla;Neimat Abd Elhakam Yassin;Aya Elsayed Maghrabia;Amira Ibrahim Morsy
Anatomy and Cell Biology
/
v.56
no.4
/
pp.526-537
/
2023
Hepatitis C virus (HCV) infection is a major health problem worldwide and its eradication is mandatory. Direct acting HCV polymerase inhibitors, such as Sofosbuvir (SOF), is an effective regimen. However, it has some side effects like mutagenesis, carcinogenesis, and the impairment of testicular function. It is important to evaluate the safety of SOF on the ovary, as there are no studies yet. Increasing the production of Reactive Oxygen Species (ROS), causes oxidative stress, which affects ovulation process, female reproduction, and fertility. Accumulation of SOF in the cells was demonstrated to promote ROS generation. Vitamin E (Vit E) is an antioxidant agent that has an essential role in the female reproductive system, its deficiency can cause infertility. We explored the effect of SOF treatment alone and co-treated with Vit E on ovarian ROS level and ovarian morphology experimentally using biochemical and immunohistochemical studies. Significant changes in oxidative stress markers; nitric oxide and malondialdehyde lipid peroxidation, antioxidant enzymes; catalase, super oxide dismutase, and reduced glutathione, proliferating markers; proliferation cell nuclear antigen and Ki-67 antigen and caspase 3 apoptotic marker were demonstrated. It was shown that where SOF induced oxidative stress, it also aggravated ovarian dysfunction. The essential role of Vit E as an antioxidant agent in protecting the ovarian tissue from the effect of oxidative stress markers and preserving its function was also displayed. This could be guidance to add Vit E supplements to SOF regimens to limit its injurious effect on ovarian function.
Purpose: Patients with hematological malignancies are at an increased risk of severe infection with coronavirus disease 2019 (COVID-19). However, developing an adequate immune response after vaccination is difficult, especially in patients with lymphoid neoplasms. Since the long-term effects of the BNT162b2 vaccine are unclear, the humoral immune response 5 months after the two vaccinations in patients with hematological disorders was analyzed. Materials and Methods: Samples were collected from 96 patients vaccinated twice with BNT162b2 and treated with at least one line of an antitumor or immunosuppressive drug in our hospital from November 2021 to February 2022. Serum anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) spike (S) antibody titers were analyzed. Patients were age- and sex-matched using propensity matching and compared with a healthy control group. Patients with serum anti-SARS-CoV-2 S antibodies were defined as 'responder' if >50 U/mL. The patients had B-cell non-Hodgkin lymphoma (B-NHL), multiple myeloma, chronic myeloid leukemia, etc. Results: Patients had significantly low antibody levels (median, 55.3 U/mL vs. 809.8 U/mL; p<0.001) and a significantly low response rate (p<0.001). Multivariate analysis showed that patients with B-NHL, aged >72 years, were associated with a low response to vaccination. There were no significant differences between patients with chronic myeloid leukemia and healthy controls. Conclusion: Our study shows that patients with hematological disorders are at risk of developing severe COVID-19 infections because of low responsiveness to vaccination. Moreover, the rate of antibody positivity differed between the disease groups. Further studies are warranted to determine an appropriate preventive method for these patients, especially those with B-NHL.
Purpose: The development and study of hepatitis C virus (HCV) vaccine candidates' individualized responses are of great importance. Here we report on an HCV DNA vaccine candidate based on selected envelope (E1/E2) epitopes. Besides, we assessed its expression and processing in human peripheral blood mononuclear cells (PBMCs) and in vivo cellular response in mice. Materials and Methods: HCV E1/E2 DNA construct (EC) was designed. The antigen expression of EC was assayed in PBMCs of five HCV-uninfected donors via a real-time quantitative polymerase chain reaction. Serum samples from 20 HCV antibody-positive patients were used to detect each individual PBMCs expressed antigens via enzyme-linked immunosorbent assay. Two groups, five Swiss albino mice each, were immunized with the EC or a control construct. The absolute count of lymph nodes' CD4+ and CD8+ T-lymphocytes was assessed. Results: Donors' PBMCs showed different levels of EC expression, ranging between 0.83-2.61-fold in four donors, while donor-3 showed 34.53-fold expression. The antigens expressed in PBMCs were significantly reactive to the 20 HCV antibody repertoire (all p=0.0001). All showed comparable reactivity except for donor-3 showing the lowest reactivity level. The absolute count % of the CD4+ T-cell significantly increased in four of the five EC-immunized mice compared to the control group (p=0.03). No significant difference in CD8+ T-cells % was observed (p=0.89). Conclusion: The inter-individual variation in antigen expression and processing dominance was evident, showing independence in individuals' antigen expression and reactivity levels to antibodies. The described vaccine candidate might result in a promising natural immune response with a possibility of CD4+ T-cell early priming.
Purpose: The purpose of this study was to evaluate the regenerative capacity of stem cells combined with bone graft material and a collagen matrix in rabbit calvarial defect models according to the type and form of the scaffolds, which included type I collagen matrix and synthetic bone. Methods: Mesenchymal stem cells (MSCs) were obtained from the periosteum of participants. Four symmetrical 6-mm-diameter circular defects were made in New Zealand white rabbits using a trephine drill. The defects were grafted with (1) group 1: synthetic bone (β-tricalcium phosphate/hydroxyapatite [β-TCP/HA]) and 1×105 MSCs; (2) group 2: collagen matrix and 1×105 MSCs; (3) group 3: β-TCP/HA, collagen matrix covering β-TCP/HA, and 1×105 MSCs; or (4) group 4: β-TCP/HA, chipped collagen matrix mixed with β-TCP/HA, and 1×105 MSCs. Cellular viability and cell migration rates were analyzed. Results: Uneventful healing was achieved in all areas where the defects were made at 4 weeks, and no signs of infection were identified during the healing period or at the time of retrieval. New bone formation was more evident in groups 3 and 4 than in the other groups. A densitometric analysis of the calvarium at 8 weeks post-surgery showed the highest values in group 3. Conclusions: This study showed that the highest regeneration was found when the stem cells were applied to synthetic bone along with a collagen matrix.
Min-Ju Kim;Ki Back Chu;Keon-Woong Yoon;Hae-Ji Kang;Dong-Hun Lee;Eun-Kyung Moon;Fu-Shi Quan
Parasites, Hosts and Diseases
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v.62
no.2
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pp.193-204
/
2024
Malaria is a global disease affecting a large portion of the world's population. Although vaccines have recently become available, their efficacies are suboptimal. We generated virus-like particles (VLPs) that expressed either apical membrane antigen 1 (AMA1) or microneme-associated antigen (MIC) of Plasmodium berghei and compared their efficacy in BALB/c mice. We found that immune sera acquired from AMA1 VLP- or MIC VLP-immunized mice specifically interacted with the antigen of choice and the whole P. berghei lysate antigen, indicating that the antibodies were highly parasite-specific. Both VLP vaccines significantly enhanced germinal center B cell frequencies in the inguinal lymph nodes of mice compared with the control, but only the mice that received MIC VLPs showed significantly enhanced CD4+ T cell responses in the blood following P. berghei challenge infection. AMA1 and MIC VLPs significantly suppressed TNF-α and interleukin-10 production but had a negligible effect on interferon-γ. Both VLPs prevented excessive parasitemia buildup in immunized mice, although parasite burden reduction induced by MIC VLPs was slightly more effective than that induced by AMA1. Both VLPs were equally effective at preventing body weight loss. Our findings demonstrated that the MIC VLP was an effective inducer of protection against murine experimental malaria and should be the focus of further development.
Min, Bo Ram;Lee, Young Mi;Park, Jae Seok;Choi, Won-Il;Kwon, Kun Young
Tuberculosis and Respiratory Diseases
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v.64
no.1
/
pp.22-27
/
2008
Background: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity. Methods: In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor. Results: The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased. Conclusion: MMPs may mediate the invasion of S. aureus into epithelial cells.
The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.
A Recent discovery of surface antigens in cells has led to the success of quantitative measurement of T-cell subpopulations, and this has especially opened the way for an epoch-making development in the understanding and classification of cellular immune mechanisms. It is known that phenotypes of T-cell subpopulations exist in many forms according to the variation of species or animal experimental models. In Korea, Clonorchis sinensis still gives rise to public concern as it infects more than eighty million people and threatens the public by causing cirrhosis of the liver, or liver cancer when liver infection becomes prolonged and chronic. Up until now there has been much progress in research and improvement in the classification system of Clonorchis sinensis in the area of humoral immunity, but as for research in the area of cellular immune mechanisms, there is almost none. Knowing all these circumstances, the authors delved for the characterization of Iymphocyte subpopulations with mice as Clonorchis sinensis in the area of cellular immunity, and obtained the following results. That is, we injected Clonorchis sinensis antigens mixed in Freund's ajuvant solution intraperitoneally in mice and measured the T-cell subpopulation characterization of spleen lymphocytes with flow cytometry. The results of these measurements showed that CD2, CD5 and CD8 decreased early following injections but then in-creased again seven weeks after the injections. CD4, however, showed a slight increase shortly after the injection but then a fair increase seven weeks after the injection.
This is the result of the annual statistic analysis of thoracic and cardiovascular surgical cases in 1992 Korea. Overall 17, 520 cases of surgery [11, 732 cases of thoracic surgery by 54 institutes / 5, 788 cases of cardiovascular surgery by 48 institutes] were done. 1. Tumor [N=2, 532] : Lung was the most frequently involved organ by tumor [54.9%],and the remainders were mediastinum [16.2%] / esophagus [14.8%] / chest wall [11.7%] / tracheobronchus [1.3%] / pleura [1.1%] in order. Of 1, 082 cases of primary lung cancer surgery,the frequency of cell type was squamous [62.6%] / adeno [21.6%] / small cell [7.1%] / large cell [2.7%]. Of 411 cases of mediastinal tumor surgery,the frequency of cell type was neurogenic [28.8%] / thymoma [27.6%] / teratoma [17.7%] / congenital cystic [17.2%]. Of 376 cases of esophageal tumor surgery,primary cancer were the most [85.4%]. 2. Infection [N=3, 157] : Pleura was the most frequently involved organ [59.0%],and the remainders were lung [31.3%] / chest wall [8.6%] / mediastinum [1.1%] in order. 3. Miscellaneous [N=6, 043] : Lung and pleural disease esp. pneumothorax [85.1%] was the most frequent surgical indication. The remainders were chest wall anomaly [3.4%] / benign esophageal disease [3.4%] / diaphragmatic pathology [2.4%] / myasthenia [1.4%] in order. Of 85 cases of thymectomy for myasthenia gravis,thymoma was noted in 58.8%. 1. Congenital heart disease [N=3, 363] : The ratio of noncyanotic to cyanotic heart disease was 3:1. Of 2, 516 cases of noncyanotic heart disease,the frequency of disease entity was VSD [44.1%] / ASD [26.0%] / PDA [19.4%] / PS [3.3%],and that of 847 cases of cyanotic heart disease was TOF [29.4%] / ECD [15.6%] / TGA [9.7%] / DORV [7.6%]. Overall mortalities were 2.1% in noncyanotic and 12.2% in cyanotic heart surgery. 2. Acquired heart disease [N=1, 929] : Of 1, 422 cases of valvular surgery,single mitral pathology was the most frequent candidate [48.0%],and total 1, 574 prosthetic valves which were mainly mechanical [95.6%] were used. Of 376 cases of coronary surgery,triple vessel was the most [35.9%],and the frequency of bypassing grafts was great saphenous vein [52.9%] / internal mammary artery [44.7%] / artificial vessel [2.4%]. Overall mortalities were 3.4% in valvular and 4.5% in coronary surgery. 3. Pericardium,Cardiac tumor,Arrhythmia,Aortic aneurysm,Assist device,and Pacemaker : There were no specific changes compared to previous survey1]. This nation-wide inquiry will be continued and reported annually by KTCS Society.
The Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) is one of the major genomic resources for human genetics and immunological studies. Use of LCLs is currently extended to pharmacogenetic studies to investigate variations in human gene expression as well as drug responses between individuals. We evaluated four common internal controls for gene expression analysis of selected hematopoietic transcriptional regulatory genes between B cells and LCLs. In this study, the expression pattern analyses showed that TBP (TATA box-binding protein) is a suitable internal control for normalization, whereas GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is not a good internal control for gene expression analyses of hematopoiesis-related genes between B cells and LCLs at different subculture passages. Using the TBP normalizer, we found significant gene expression changes in selected hematopoietic transcriptional regulatory genes (downregulation of RUNX1, RUNX3, CBFB, TLE1, and NOTCH2 ; upregulation of MSC and PLAGL2) between B cells and LCLs at different passage numbers. These results suggest that these hematopoietic transcriptional regulatory genes are potential cellular targets of EBV infection, contributing to EBV-mediated B-cell transformation and LCL immortalization.
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