Mitochondrial damage induced by HTLV-1 infection in host cells / 中华微生物学和免疫学杂志

Objective@#To investigate the effects of human adult T lymphoblastic leukemia virus typeⅠ (HTLV-1) infection on the production of reactive oxygen species (ROS) and mitochondrial damage in host cells.@*Methods@#A cell model of HTLV-1 infection was established by co-culturing HTLV-1-positive cell line MT2 with HeLa cells. ROS, mitochondrial membrane potential (MMP) and total mitochondria were detected using specific fluorescence probe labeling method. Cell apoptosis was detected by Annexin V-FITC/PI method. Western blot was performed to detect viral proteins Tax and p19, as well as mitochondrial proteins TIM23 and TOM20. After the treatment of MT2 cells with different concentrations of reverse transcription inhibitors (ZDV), relative viral loads were detected by quantitative real-time PCR and Western blot, and the mass of mitochondria was analyzed by flow cytometry.@*Results@#After co-culturing HeLa cells with MT2 cells for 24 h, the ROS level in host cells increased without obvious cell apoptosis, while the mitochondrial membrane potential, mitochondrial protein expression and total mitochondria decreased significantly. When the replication of HTLV-1 in MT2 cells was inhibited by ZDV, the ROS level and total mitochondria increased.@*Conclusions@#HTLV-1 infection can cause oxidative stress in host cells, resulting in mitochondrial damage. Autophagy might be activated to degrade mitochondrial damage and maintain cell homeostasis during the infection.

Mitochondrial damage induced by HTLV-1 infection in host cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of human adult T lymphoblastic leukemia virus typeⅠ (HTLV-1) infection on the production of reactive oxygen species (ROS) and mitochondrial damage in host cells. Methods A cell model of HTLV-1 infection was established by co-culturing HTLV-1-positive cell line MT2 with HeLa cells. ROS, mitochondrial membrane potential ( MMP) and total mitochondria were detected using specific fluorescence probe labeling method. Cell apoptosis was detected by Annexin V-FITC/PI method. Western blot was performed to detect viral proteins Tax and p19, as well as mitochondrial pro-teins TIM23 and TOM20. After the treatment of MT2 cells with different concentrations of reverse transcrip-tion inhibitors ( ZDV) , relative viral loads were detected by quantitative real-time PCR and Western blot, and the mass of mitochondria was analyzed by flow cytometry. Results After co-culturing HeLa cells with MT2 cells for 24 h, the ROS level in host cells increased without obvious cell apoptosis, while the mitochon-drial membrane potential, mitochondrial protein expression and total mitochondria decreased significantly. When the replication of HTLV-1 in MT2 cells was inhibited by ZDV, the ROS level and total mitochondria increased. Conclusions HTLV-1 infection can cause oxidative stress in host cells, resulting in mitochon-drial damage. Autophagy might be activated to degrade mitochondrial damage and maintain cell homeostasis during the infection.

Growth of the human lens in the Indian adult population: Preliminary observations.

Context: The eye lens grows throughout life by the addition of new cells inside the surrounding capsule. How this growth affects the properties of the lens is essential for understanding disorders such as cataract and presbyopia. Aims: To examine growth of the human lens in the Indian population and compare this with the growth in Western populations by measuring in vitro dimensions together with wet and dry weights. Settings and Design: The study was conducted at the research wing of a tertiary eye care center in South India and the study design was prospective. Materials and Methods: Lenses were removed from eye bank eyes and their dimensions measured with a digital caliper. They were then carefully blotted dry and weighed before being placed in 5% buffered formalin. After 1 week fixation, the lenses were dried at 80 °C until constant weight was achieved. The constant weight was noted as the dry weight of the lens. Statistical Analysis Used: Lens parameters were analyzed as a function of age using linear and logarithmic regression methods. Results: Data were obtained for 251 lenses, aged 16–93 years, within a median postmortem time of 22 h. Both wet and dry weights increased linearly at 1.24 and 0.44 mg/year, respectively, throughout adult life. The dimensions also increased continuously throughout this time. Conclusions: Over the age range examined, lens growth in the Indian population is very similar to that in Western populations.

Adult Reye-like syndrome associated with serologic evidence of acute parvovirus B19 infection

Reye's syndrome is an infrequently diagnosed medical condition affecting mainly children. The etiology, epidemiology and natural history of Reye's syndrome have been cloudily written in footnotes of medical books and exotic papers since the initial description in early 1950s. We report here a case of adult Reye's syndrome associated with serologic evidence of parvovirus B19 infection.

MHC Antigen Expressions in Human Embryonic Neural Stem Cells and Adult Breast Epithelial Stem Cells / 대한이식학회지

PURPOSE: Due to their unique capacity to self-renew and for multiple differentiation, stem cells are considered potent candidates for cell replacement therapy in many devastating diseases. However, studies on immune rejection, which is a major problem facing successful stem cell therapy, are rare. Thus, we examined MHC expression of human stem cells and effects of IFN-gamma on the MHC class I expression of the cells in order to determine whether human stem cells might be rejected after transplantation. METHODS: The MHC antigen expressions of human embryonic neural stem cell line (HB1.F3) and human breast epithelial stem cell line (M13SV1) were examined by RT-PCR and FACS. The effects of varying concentrations of IFN-gamma and of varying incubation times with IFN-gamma on the expression of MHC class I antigens in these stem cell lines were also examined by FACS. RESULTS: The results show low expression levels of MHC class I antigens on surfaces of these cells. A dramatic induction of MHC class I expression was observed when the cells were treated with IFN-gamma. Maximal induction of MHC class I antigen expression in HB1.F3 and M13SV1 cells was observed at above the concentrations of 20 ng/mL and 5 ng/mL of IFN-gamma 48 h after treatment, respectively. Elevated MHC class I levels in HB1.F3 and M13SV1 cells were sustained for 48 h and 72 h after withdrawing IFN-gamma, respectively. CONCLUSION: These results suggest that human stem cells express high levels of MHC class I antigens, and thus may be rejected on transplantation unless they are modified. Therefore, in addition to studies on stem cell differentiation, studies on overcoming the immunological barriers to stem cell transplantation are prerequisite for successful clinical application of stem cell therapy.

The Establishment of the Apoptotic Model of Cultured Human Adult Cardiomyocytes / 中国医师杂志

Objective To establish a model for hypoxia-induced apoptosis of human adult cardiomyocytes in vitro. Methods The human adult heart cells of ventricular muscle were isolated by the digestion of trypsin and collagenase, cultured in Iscove's modified Dulbecco's medium (IMDM), and purified by means of differential attachment technique, adding Brdu and slicing fibroblasts by sterile needle. After 4 to 6 days, the cardiomyocytes were cultured in an incubator containing 3% O 2, 92% N 2 and 5% CO 2 for 6, 12, 24 and 48h to form hypoxia injury for cells. Morphological changes of apoptotic cardiomyocytes were measured by invertion microscopy and electronic microscopy. Apoptotic rate was measured by TUNEL staining. Results The cardiomyocytes occurred opoptosis after hypoxia injury, apoptotic rate of which at 6, 12, 24 and 48h after hypoxia was (3 3?0 9)%, (8 3?1 8)%, (16 1?2 6)% and (19 4?2 3)% respectively. Conclusion The apoptotic rate of cardiomyocytes increased with time of hypoxia lengthened. The model for hypoxia-induced apoptosis of human adult cardiomyocytes was successful and reliable.

Light and Electron Microscopic Study on Metaplasia of Epithelium of Esophago-gastric Junction in Human Adult / 대한해부학회지

Columnar epithelium-lined esophagus or intestinal metaplasia of the stomach are acquired disorder associated with some pathologic conditions such as reflux esophagitis and gastric carcinoma. The aims of this study were to investigate the characteristics of the metaplastic epithelium in 12 resected specimens of esophago-gastric junction (EGJ) from the patients of gastric carcinoma by histochemistry, immunohistoche-mistry, and electron microscopy. By hematoxylin-eosin staining, metaplasia was found in all 12 cases examined, among them 10 specimens were intestinal metaplasia of the gastric epithelium, 2 cases columnar epithelium-lined esophagus. Intestinal metaplasia was villiform and consisted of columnar cells and mucous (goblet) cells, and showed variations in the pattern of arrangement: metaplasia occurred in EGJ region, occurred in the gastric epithelium apart some distance from EGJ, intermingled with gastric epithelium. The mucous cells of the intestinal metaplasia were stained with both PAS and alcian blue. The gastric epithelium, however, was only stained with PAS. The columnar epithelium-lined esophagus was stained with PAS but not stained with alcian blue. This simple epithelium was positive for 5D3. Interestingly in 2 cases, stratified squamous epithelium was interposed between gastric epithelium. This epithelium was positive for AE8 and was distinguished from gastric epithelium which were positive for 5D3. The columnar cells of the intestinal metaplasia showed regular microvilli, many apical mitochondria, and well developed supranuclear Golgi complex which were characteristic features of typical absorptive cells. The mucous cells were oval in shape and contained numerous mucous droplets in the apical cytoplasm. Intestinal gland consisted of undifferentiated cells and mucous cells, and did not have Paneth cell. Gastric gland consisted of mucous cells of similar fine structural features to those of gastric epithelial cell. Chief cell and parietal cell were not present in the glandular epithelium. These results indicate that most of the metaplasia occurs in the gastic cardiac portion rather than esophageal portion in EGJ, and suggest that the histologic type of metaplsia of gastric epithelium is complete, and that of esophageal epithelium is junctional.