Real-time observation of neutrophil extracellular trap formation in the inflamed mouse brain via two-photon intravital imaging / 한국실험동물학회지

Intravital imaging via two-photon microscopy (TPM) is a useful tool for observing and delineating biological events at the cellular and molecular levels in live animals in a time-lapse manner. This imaging method provides spatiotemporal information with minimal phototoxicity while penetrating a considerable depth of intact organs in live animals.Although various organs can be visualized using intravital imaging, in the field of neuroscience, the brain is the main organ whose cell-to-cell interactions are imaged using this technique. Intravital imaging of brain disease in mouse models acts as an abundant source of novel findings for studying cerebral etiology. Neutrophil infiltration is a wellknown hallmark of inflammation; in particular, the crucial impact of neutrophils on the inflamed brain has frequently been reported in literature. Neutrophil extracellular traps (NETs) have drawn attention as an intriguing feature over the last couple of decades, opening a new era of research on their underlying mechanisms and biological effects.However, the actual role of NETs in the body is still controversial and is in parallel with a poor understanding of NETs in vivo. Although several experimental methods have been used to determine NET generation in vitro, some research groups have applied intravital imaging to detect NET formation in the inflamed organs of live mice. In this review, we summarize the advantages of intravital imaging via TPM that can also be used to characterize NET formation, especially in inflamed brains triggered by systemic inflammation. To study the function and migratory pattern of neutrophils, which is critical in triggering the innate immune response in the brain, intravital imaging via TPM can provide new perspectives to understand inflammation and the resolution process.

Three-Dimensional Distribution of Cochlear Macrophages in the Lateral Wall of Cleared Cochlea

Objectives@#. Resident macrophages are well known to be present in the cochlea, but the exact patterns thereof in spiral ligaments have not been discussed in previous studies. We sought to document the distribution of macrophages in intact cochleae using three-dimensional imaging. @*Methods@#. Cochleae were obtained from C-X3-C motif chemokine receptor 1+/GFP mice, and organ clearing was performed. Three-dimensional images of cleared intact cochleae were reconstructed using two-photon microscopy. The locations of individual macrophages were investigated using 100-μm stacked images to reduce bias. Cochlear inflammation was then induced by lipopolysaccharide (LPS) inoculation into the middle ear through the tympanic membrane. Four days after inoculation, three-dimensional images were obtained. @*Results@#. Macrophages were scarce in areas adjacent to the stria vascularis, particularly the area just beneath it even though many have suspected macrophages to be abundant in this area. This finding remained consistent upon LPS-induced cochlear inflammation, despite a significant increase in the number of macrophages, compared to non-treated cochlea. @*Conclusion@#. Resident macrophages in spiral ligaments are scarce in areas adjacent to the stria vascularis.

Three-Dimensional Distribution of Cochlear Macrophages in the Lateral Wall of Cleared Cochlea

Objectives@#. Resident macrophages are well known to be present in the cochlea, but the exact patterns thereof in spiral ligaments have not been discussed in previous studies. We sought to document the distribution of macrophages in intact cochleae using three-dimensional imaging. @*Methods@#. Cochleae were obtained from C-X3-C motif chemokine receptor 1+/GFP mice, and organ clearing was performed. Three-dimensional images of cleared intact cochleae were reconstructed using two-photon microscopy. The locations of individual macrophages were investigated using 100-μm stacked images to reduce bias. Cochlear inflammation was then induced by lipopolysaccharide (LPS) inoculation into the middle ear through the tympanic membrane. Four days after inoculation, three-dimensional images were obtained. @*Results@#. Macrophages were scarce in areas adjacent to the stria vascularis, particularly the area just beneath it even though many have suspected macrophages to be abundant in this area. This finding remained consistent upon LPS-induced cochlear inflammation, despite a significant increase in the number of macrophages, compared to non-treated cochlea. @*Conclusion@#. Resident macrophages in spiral ligaments are scarce in areas adjacent to the stria vascularis.

Mitofusin-2 Promotes the EpithelialMesenchymal Transition-Induced Cervical Cancer Progression

High expression of mitofusin-2 (MFN2), a mitochondrial fusion protein, has been frequently associated with poor prognosis of patients with cervical cancer. Here, we aimed to identify the function of MFN2 in cervical cancer to understand its influence on disease prognosis.To this end, from cervical adenocarcinoma, we performed an MTT assay and quantitative RT-PCR (qRT-PCR) analysis to assess the effects of MFN2 on the proliferation and of HeLa cells. Then, colony-formation ability and tumorigenesis were evaluated using a tumor xenograft mouse model. The migration ability related to MFN2 was also measured using a wound healing assay. Consequently, epithelial-mesenchymal transition (EMT) of MFN2-knockdowned HeLa cells originating from adenocarcinoma. markers related to MFN2 were assessed by qRT-PCR. Clinical data were analyzed using cBioPortal and The Cancer Genome Atlas. We found that MFN2 knockdown reduced the proliferation, colony formation ability, migration, and in vivo tumorigenesis of HeLa cells. Primarily, migration of MFN2-knockdowned HeLa cells decreased through the suppression of EMT. Thus, we concluded that MFN2 facilitates cancer progression and in vivo tumorigenesis in HeLa cells. These findings suggest that MFN2 could be a novel target to regulate the EMT program and tumorigenic potential in HeLa cells and might serve as a therapeutic target for cervical cancer. Taken together, this study is expected to contribute to the treatment of patients with cervical cancer.

Neutrophil Extravasation Cascade: What Can We Learn from Two-photon Intravital Imaging?

Immune cells (leukocytes or white blood cells) move actively and sensitively based on body conditions. Despite their important role as protectors inside the body, it is difficult to directly observe the spatiotemporal momentum of leukocytes. With advances in imaging technology, the introduction of two-photon microscopy has enabled researchers to look deeper inside tissues in a three-dimensional manner. In observations of immune cell movement along the blood vessel, vascular permeability and innate immune cell movements remain unclear. Here, we describe the neutrophil extravasation cascade, which were observed using a two-photon intravital imaging technique. We also provide evidence for novel mechanisms such as neutrophil body extension and microparticle formation as well as their biological roles during migration.

Extravasating Neutrophil-derived Microparticles Preserve Vascular Barrier Function in Inflamed Tissue

Emerging evidence suggests that gap formation and opening of the endothelial junctions during leukocyte extravasation is actively controlled to maintain the integrity of the vascular barrier. While the role for endothelial cells to this process has been well defined, it is not clear whether leukocytes are also actively contributing to endothelial barrier function. We have recently showed that extravasating leukocytes deposit microparticles on the subendothelium during the late stages of extravasation, which is LFA-1 dependent. Using multiphotonintravital microscopy (MP-IVM) of mouse cremaster muscle vessels in the current work, we show that microparticle formation and deposition maintains the integrity of the microvascular barrier during leukocyte extravasation. Inhibition of neutrophil-derived microparticle formation resulted in dramatically increased vascular leakage. These findings suggest that deposition of microparticles during neutrophil extravasation is essential for maintaining endothelial barrier function and may result in temporal difference between neutrophil extravasation and an increase in vascular leakage.

Symptomatic Analysis of Men with Chronic Prostatitis/Chronic Pelvic Pain Syndrome

PURPOSE: Prostatitis is an extremely common syndrome that afflicts 2~10% of men and is now classified into NIH category. Of these, NIH category III(Chronic Prostatitis/Chronic Pelvic Pain Syndrome, CP/CPPS) varies widely in clinical presentation and response to treatment. We evaluated the characteristic symptoms of CP/CPPS based on NIH chronic prostatitis symptom index(NIH-CPSI), International Prostate Symptom Score(IPSS) and International Index of Erectile Function-5(IIEF-5) for the exploration of the future prospects of treatment. MATERIALS AND METHODS: 114 subjects agreed to participate in the study. The data from 99 patients were available for evaluation. The candidates with category IIIa and IIIb chronic pelvic pain syndrome were from 3rd decade to 6th decade for exclusion of effect of BPH. The NIH-CPSI, IPSS, IIEF-5 were used to grade symptoms and the quality of life impact at the start of the study. We investigated the correlation of the each domain of the scales categorized by age, location and multiplicity of the pain, and duration of the disease. Pearson's correlation method and ANOVA test were used for statistical analysis. RESULTS: Mean age of the patients was 41.1+/-8.5(3rd decade; 8 patients, 4th decade; 30, 5th decade 47, 6th decade 14). There were significant correlation between IPSS total score and urinary domain of NIH-CPSI (R=0.82), and irritative domain of IPSS and urinary domain of NIH-CPSI(R=0.76) with pearson's method. Analysis of the scores categorized by age revealed that there were no difference in 2nd, 3rd, 4th decade, but 6th decades had high IPSS total score. Analysis of the scores categorized by location and multiplicity of the pain revealed that the most common site of the pain was perineum(34.3%), and the patients who had 3 or more painful area had higher scores in all domains of CPSI and IPSS except pain domain of NIH-CPSI than the patients who had 1 painful area. Analysis of the scores categorized by duration of the disease revealed that the IPSS scores of the patients suffered during 1 year or more were higher than the patient suffered during 6months or less. There was no significant correlation between IIEF-5 and other scales. CONCLUSION: In our study there was significant correlation between urinary domain of NIH-CPSI and total score of IPSS. So, urinary domain of NIH-CPSI could be helpful index when alpha-blocker is used for the treatment of CP/CPPS. And alpha-blocker could be more effective in patients who are older and have longer duration of the disease and have multiple painful areas.

Comparison of Waters' radiography, panoramic radiography, and computed tomography in the diagnosis of antral mucosal thickening / 대한구강악안면방사선학회지

With the CT findings as gold standard, the sensitivity, the specificity, and the diagnostic accuracy of WatersO radiography and WatersO radiography with panoramic radiography were compared in the diognosis of antral mucosal thickening of 16 patients. Three oral radiologists and three non-oral radiologists interpreted the WatersO radiographs and after 4 weeks, interpreted the WatersO radiographs and panoramic radiographs simutaneously. The interpretation was the existence or the non-existence of the mucosal thickening on the medial, the posterolateral, the floor, and the roof of maxillary sinus. The obtained results were as followed ; 1. In oral radiologist group, the sensitivity, specificity and diagnostic accuracy of WatersO film were 0.7250, 0.8489 and 0.7578 respectively. 2. The sensitivity and the diagnostic accuracy in oral radiologist group were higher than those of non-oral radiologist group (P0.05). 3. There was no significant difference of the diagnostic abilities except the specificity in oral radiologist group between WatersO radiography and WatersO radiography with panoramic radiography (p>0.05). 4. The sensitivity and the diagnostic accuracy were the highest in the case of medial wall interpretation, the specificity was the highest in the posterioateral wall. 5. In the posterolateral wall and the floor, the sensiticity and the diagnostic accuracy of oral radiologist group were higher than those of non-oral radiologist group (P<0.05).

Nonspecific inflammation in the face / 대한구강악안면방사선학회지

Patient with compalints of swelling, pain in the maxillary region and discomfort visited Seoul National University Dental Hospital in August last year. Clinical examination and diagnostic imagings implied he was suffered from fungal h yphal infection but no causative fungus was found by the histopathologic and microbiologic investigation. Therefore he w as diagnosed with nonspecific inflammation. But as yet, we do think this case is very similar to some kinds of mucomycos is. So we presented this case for more thorough discussion. Following are founded in the examination. 1. Patient had sufferd from Diabetes mellitus and complained of stuffness, headache, swelling in buccal cheeks and pare sthesia. And we found more maxillary bony destruction and ulcer with elevated margin in the palate by clinical examinati on. 2. In the first visit, Plain films revealed general bony destruction of the maxilla, radiopaqueness in the sinonasal ca vities. CT and MRI showed soft tissue mass filled in the paranasal sinus except frontal sinus and bony destruction in in volved bones. 3. No causative bacteria and fungus was found in the biopsy and microbiologic cultures. 4. Caldwell-Luc operation and curettage were carried and antibiotics were taken for 4 months. But now he was worse than in the past. 5. In the second visit, involvement of orbit, parapharyngeal sinus, clivus, cavernous sinus and middle cranial fossa we re seen clearly in the CT and MRI.

Nonspecific inflammation in the face / 대한구강악안면방사선학회지

Patient with compalints of swelling, pain in the maxillary region and discomfort visited Seoul National University Dental Hospital in August last year. Clinical examination and diagnostic imagings implied he was suffered from fungal h yphal infection but no causative fungus was found by the histopathologic and microbiologic investigation. Therefore he w as diagnosed with nonspecific inflammation. But as yet, we do think this case is very similar to some kinds of mucomycos is. So we presented this case for more thorough discussion. Following are founded in the examination. 1. Patient had sufferd from Diabetes mellitus and complained of stuffness, headache, swelling in buccal cheeks and pare sthesia. And we found more maxillary bony destruction and ulcer with elevated margin in the palate by clinical examinati on. 2. In the first visit, Plain films revealed general bony destruction of the maxilla, radiopaqueness in the sinonasal ca vities. CT and MRI showed soft tissue mass filled in the paranasal sinus except frontal sinus and bony destruction in in volved bones. 3. No causative bacteria and fungus was found in the biopsy and microbiologic cultures. 4. Caldwell-Luc operation and curettage were carried and antibiotics were taken for 4 months. But now he was worse than in the past. 5. In the second visit, involvement of orbit, parapharyngeal sinus, clivus, cavernous sinus and middle cranial fossa we re seen clearly in the CT and MRI.