Single-cell RNA sequencing identifies distinct transcriptomic signatures between PMA/ionomycin- and αCD3/αCD28-activated primary human T cells

Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell–derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55+) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

Restoration of Immune Privilege in Human Dermal Papillae Controlling Epithelial-Mesenchymal Interactions in Hair Formation

BACKGROUND@#Hair follicles are among a handful of organs that exhibit immune privilege. Dysfunction of the hair follicle immune system underlies the development of inflammatory diseases, such as alopecia areata. @*METHODS@#Quantitative reverse transcription PCR and immunostaining was used to confirm the expression of major histocompatibility complex class I in human dermal papilla cells. Through transcriptomic analyses of human keratinocyte stem cells, major histocompatibility complex class I was identified as differentially expressed genes. Organ culture and patch assay were performed to assess the ability of WNT3a conditioned media to rescue immune privilege. Lastly, CD8? T cells were detected near the hair bulb in alopecia areata patients through immunohistochemistry. @*RESULTS@#Inflammatory factors such as tumor necrosis factor alpha and interferon gamma were verified to induce the expression of major histocompatibility complex class I proteins in dermal papilla cells. Additionally, loss of immune privilege of hair follicles was rescued following treatment with conditioned media from outer root sheath cells. Transcriptomic analyses found 58 up-regulated genes and 183 down-regulated genes related in MHC class I? cells. Using newborn hair patch assay, we demonstrated that WNT3a conditioned media with epidermal growth factor can restore hair growth. In alopecia areata patients, CD8? T cells were increased during the transition from mid-anagen to late catagen. @*CONCLUSION@#Identification of mechanisms governing epithelial and mesenchymal interactions of the hair follicle facilitates an improved understanding of the regulation of hair follicle immune privilege.

Fruit and vegetable consumption frequency and mental health in Korean adolescents: based on the 2014–2017Korea Youth Risk Behavior Survey / 한국영양학회지

Purpose@#This study examined the association between fruit and vegetable intake and mental health in Korean adolescents. @*Methods@#This study used the data from the 2014–2017 Korea Youth Risk Behavior Survey, a national cross-sectional survey on 137,101 boys and 130,806 girls aged 12–18. Fruit and vegetable intake was assessed based on the frequency of consumption. The outcome variables were the perceived happiness, perceived stress, depressive symptom and suicidal ideation over the previous 12 months. Logistic regression models were used after adjusting for the demographic, life style and other dietary factors. @*Results@#Only 34% and 29% of Korean adolescents consumed fruits more than 5 times/ week and vegetables more than 2 times/day, respectively; whereas 37%, 25% and 12.2% of Korean adolescents had perceived stress, depressive symptom and suicidal ideation, respectively. After adjusting for the confounding variables, the greater consumption of fruit and vegetable were all associated with a higher odds of perceived happiness; the adjusted odds ratios (AORs) (95% CI) were 1.53 (1.46–1.60) in boys and 1.82 (1.73–1.90) in girls who consumed fruit ≥ 5 times/week, and 1.65 (1.54–1.76) in boys and 1.62 (1.51–1.72) in girls who consumed vegetable ≥ 2 times/day. In contrast, the consumption of fruit or vegetable were all significantly associated with a lower odds of perceived stress, depressive symptom, and suicidal ideation; the AOR (95% CI) were 0.70 (0.67–0.73), 0.88 (0.84–0.93), and 0.78 (0.73– 0.83) in boys who consumed fruit 3–4 times/week, and 0.71 (0.67–0.76), 0.88 (0.81–0.94), and 0.68 (0.62–0.74) in boys who consumed vegetable 5–7 times/week. Similar associations of fruit or vegetable consumption with perceived stress, depressive symptom, or suicidal ideation were found in girls. @*Conclusion@#These findings provide evidence that increasing fruit and vegetable intake is important for better mental health among adolescents.

Modified Transosseous Wiring Technique for Neglected Fracture–Dislocation of the Proximal Interphalangeal Joint

BACKGROUND: Fracture–dislocation of the proximal interphalangeal (PIP) joint of the finger is challenging due to the high risk of stiffness. The purpose of this study is to evaluate the clinical and radiological results of a modified transosseous wiring technique for the management of chronic fracture–dislocations of the PIP joint. METHODS: Ten patients (nine men and one woman; mean age, 38.3 years; range, 21 to 69 years) with neglected fracture–dislocation of the PIP joint were included. The mean duration from injury to operation was 14.7 weeks (range, 3 to 66 weeks). The dorsolateral approach and extension block pinning were used to reduce dislocation. After thorough debridement of the scar tissues in the dorsal dead space and the fracture site, the reduction was maintained with transosseous wiring. Radiologic evaluations of bone union and arthritic changes and clinical evaluations (range of motion of the PIP joint and Disabilities of the Arm, Shoulder and Hand [DASH] score) were performed. The mean follow-up period was 12.9 months (range, 12 to 19 months). RESULTS: All patients demonstrated evidence of radiographic healing within a mean healing time of 6 weeks (range, 4 to 10 weeks); however, one had a widened gap and one had an early arthritic change. The mean range of motion in the PIP joint was 81° (range, 50° to 105°). The mean DASH score was 21.6 (range, 7.5 to 35.8). CONCLUSIONS: For chronic fracture–dislocation of the PIP joint, transosseous wiring with direct curettage and optimal bone purchase can provide satisfying outcome.

Extensor Pollicis Longus Tendon Rupture with Concomitant Rupture of the Extensor Digitorum Communis II Tendon and Extensor Indicis Proprius after Volar Plating for Distal Radius Fracture

Extensor tendon rupture is well known complication following distal radius fracture after either conservative treatment or volar plating. However, there are not many reports in literature about concomitant ruptures of other extensor tendons. We report a case of delayed rupture of extensor pollicis longus (EPL), second extensor digitorum communis (EDC II), and extensor indicis proprius (EIP) tendons 4 weeks after volar plating for distal radius fracture. Due to the absence of EIP, EIP transfer was discouraged for EPL reconstruction. Thumb and index finger extension was restored by palmaris longus tendon graft for EPL and EDC II.

Signature Genes in Macrodactyly through Transcriptome Network Analysis Reveal their Association of Lipid Metabolism / 체질인류학회지

Macrodactyly is one of the most difficult hand anomalies to treat not only surgically but medically as well. Little is known about the molecular pathways and lipid metabolism of this disease. To elucidate the potential mechanism of macrodactyly progress, we used the bioinformatical analysis including quantile normalization, principal component analysis, heatmap and volcano plot. For the functional bioinformatical study, lipid, lipoprotein and phospholipid metabolism of Kyoto Encyclopedia of Genes and Genomes, Wiki Pathways, and Reactome Pathway were utilized to compare the differentially expressed genes in macrodactyly with control group. We found up-regulation of CDK6 and E2F1, which are associated with the mitotic cell cycle of cancer cells. PIK3CG, associated with cancer and lipid metabolism, was also enriched in macrodactyly. In down-regulated genes, PTEN was highlighted in lipid metabolism, phosphatidylinositol signaling system and insulin signaling. ABCD3, related in peroxisomal import of fatty acids, was also down-regulated. In this study, we predicted the pathogenic candidate genes as well as the potential molecular pathways related to macrodactyly by identifying the signature genes. Signature genes through systems bioinformatical analysis can be utilized to catch the insight of the molecular pathogenesis of macrodactyly.

Comparative Analyses of Signature Genes in Acute Rejection and Operational Tolerance

Using biomarkers as prediction tools or therapeutic targets can be a valuable strategy in transplantation. Recent studies identified biomarkers of acute rejection (AR) and operational tolerance (TOL) through the application of meta-analysis. In this study, we comparatively analyzed the signature genes in acute rejection and operational tolerance seen in human allogeneic transplantations using massive bioinformatical meta-analysis. To identify the signature genes in opposite immunological conditions, AR and TOL, we first collected the 1,252 gene expression data specifically intended for those circumstances. Then we excluded based on biological cut-values, Principal Component Analysis (PCA) as well as Multi-Dimensional Scaling (MDS). Using differentially expressed genes (DEGs) from meta-analysis, we then applied a ranked scoring system to identify the signature genes of AR and TOL. We identified 53 up-regulated and 32 down-regulated signature genes in acute rejection condition. Among them, ISG20, CXCL9, CXCL10, CCL19, FCER1G, PMSE1, UBD are highly expressed in AR condition. In operational tolerance, we identified 110 up-regulated and 48 down-regulated signature genes. TCL1A, BLNK, MS4A1, EBF1, IGHM are up-regulated in TOL condition. These genes are highly representative of AR or TOL across the different organs such as liver, kidney and heart. Since immune response is the sum of complex biological and molecular dynamics, these signature genes as well as pathway analysis using a systems biology approach could be used to catch the insights of the certain pathways that would be overlooked with the conventional gene-level comparative analysis.

Gene Expression Analysis in Nasal Polyp Using Microarray / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: A nasal polyp is a chronic inflammatory disease that affects 1~4% of the general adult population. It is characterized by edematous inflamed mucosa prolapsing into the nose, leading to nasal obstruction, abnormal secretions, loss of smell, headache, and a reduced quality of life. We sought to screen for diseases related genes in nasal polyp by using DNA microarray and to validate the altered expression of selected genes. SUBJECTS AND METHOD: The samples were obtained from nine patients with nasal polyps and two patients with pituitary adenoma. DNA microarrays containing 22,063 genes were used to compare individual gene profiles of nasal polyp samples and normal mucosal samples. A study of mRNA expression of selected genes was performed using RT-PCR. RESULTS: 126 genes were up-regulated at least 2-fold, and 161 genes were down-regulated at least 50% in nasal polyps compared with the normal sinus mucosa. The three most up-regulated genes (STATH, TCN1, DMBT1) and one most down-regulated gene (SCGB1A1) were retested by means of RT-PCR. RT-PCR confirmed the differential expressions of all the genes in nasal polyp compared with normal sinus mucosa. CONCLUSION: This study could provide a clue for investigating possible pathophysiologic processes involved in nasal polyps.

Difference of Gene Expression between Hypertrophic Scar Keratinocytes and Normal Keratinocytes

PURPOSE: There is no clear evidence of the original cause of hypertrophic scar, and the effective method of treatment is not yet established. Recently the steps of searching in gene and molecular level are proceeding. we are trying to recognize the difference between keratinocytes of hypertrophic scar and normal skin. Then we do support the comprehension of the scar formation mechanism and scar management. METHODS: Total RNAs were extracted from cultured keratinocytes from 4 hypertrophic scars and normal skins. The cDNA chips were prepared. A total of 3063 cDNAs from human cDNA library were arrayed. And the scanning data were analyzed. RESULTS: On microarray, heat shock protein, pyruvate kinase, tumor rejection antigen were more than 2 fold intensity genes. Among them, heat shock 70 kd protein showed the strongest intensity difference. CONCLUSION: In this study, it can be concluded that heat shock proteins play an important role in the process of wound healing and scar formation. This study provides basic biologic information for scar research. The new way of the prevention and treatment of scar formation would be introduced with further investigations.

The Usefulness of Standard Electroretinogram in Classification of Central Retinal Vein Occlusion

PURPOSE: Central retinal vein occlusion is classified as ischemic and non-ischemic types. Ischemic type has poor visual prognosis because of neovascular complications. Fluorescein angiography shows lower reproducibility and predictability of neovascularization and has complications although it has been the standard method for classification. This study was made to determine the usefulness of the electroretinogram (ERG) in the classification and predictor of neovascularization. METHODS: Forty-two eyes with central retinal vein occlusion were classified as ischemic and non-ischemic groups by means of indirect ophthalmoscope and fluorescein angiography finding. All patients were examined with standard electroretinogram and followed. During the follow-up period, development of neovascularization was evaluated. All parameters of electroretinogram were analyzed. RESULTS: Compared to the control group, a and b wave amplitudes in maximal combined response, b/a ratio, b wave amplitude in cone response, and b wave implicit time in 30Hz flicker response showed statistically significant difference in only ischemic central retinal vein occlusion (P<0.05). Compared to the control group, a and b wave amplitudes in maximal combined response, b/a ratio show statistically significant difference in only central retinal vein occlusion with rubeosis. CONCLUSIONS: Standard electroretinogram is thought to be a useful method for classifying the central retinal vein occlusion and predicting the rubeosis in central retinal vein occlusion.

Transpupillary Thermotherapy in Circumscribed Choroidal Hemangioma

PURPOSE: We described a patient with decreased vision due to circumscribed choroidal hemangioma and the patient was treated with transpupillary thermotherapy. METHODS: A 40-year-old male with circumscribed choroidal hemangioma combined with serous retinal detachment involving macula was managed by 2 sessions of transpupillary thermotherapy with 810 mm infrared diode laser. RESULTS: Improvement of central visual acuity, atrophy of circumscribed choroidal hemangioma and reabsorption of serous retinal detachment were observed without any complication during 6 months follow up period after 2 sessions of transpupillary thermotherapy. CONCLUSIONS: Transpupillary thermotherapy is a safer and alternatively effective treatment in circumscribed choroidal hemangioma with serous retinal detachment rather than ordinary photocoagulation or radiotherapy.

A case of idiopathic thrombocytopenic purpura / 대한피부과학회지

Idiopathic thrombocytupenic purpura is known as an autommune t,hrombocytopenic purpura. It is characterized by either an acute or gradual onset of petechiae or ecchymoses in the skin and mucous membranes, especially in the mouth. Epistaxis, conjunct.ival hemorrhage, hemorrhagic bullae in the mouth, gingival bleeding, melena, and hematemesis may occur. We report a case of idiopathic thrombocytopenic purpura in a 10-year-old male, who showed multiple petechiie and ecchymoses on both lower extremities, and hemorchagic bullae in the mouth. Periheral blood smear showed decreased platlet count, and in the bone marrow megakeyocytes were incraesed. Bleeding time was increased, but coagulation time was normal.